Automated embryonic axis extraction device

Abstract

Apparatus(es), system(s), and method(s) provide for automated separation / extraction and collection of embryonic axes from dicot seeds. Such apparatus(es), system(s), and method(s) can include a rotating suspension of a combination of whole dicot seeds and liquid, wherein the whole dicot seeds are comminuted to substantially separate / extract the embryonic axes. The apparatus(es), system(s), and method(s) can further include through-pores wherein the separated / extracted embryonic axes are generally passed through the through-pores for collection. Embryonic axes separated from seeds can be used for genetic research and experimentation including, but not limited to, biolistic transformation(s), biolistic gene delivery, agrobacterial transformation(s), and / or gene editing.

Description

Agent Ref. No. P14771WOOOTITLE : AUTOMATED EMBRYONIC AXIS EXTRACTION DEVICECROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit under 35 U.S.C. § 119 to provisional patent application U.S. Serial No. 63 / 709,108, filed October 18, 2024. The provisional patent application is herein incorporated by reference in its entirety, including without limitation, the specification, claims, and abstract, as well as any figures, tables, appendices, or drawings thereof.TECHNICAL FIELD

[0002] The present disclosure relates generally to apparatus(es), method(s), and / or system(s) having applications in at least the genetic editing, genetic modification, plant modification, and agriculture industries. More particularly, but not exclusively, the present disclosure relates to apparatus(es), method(s), and / or system(s) for automatically extracting embryonic axes from plant seeds.BACKGROUND

[0003] The background description provided herein gives context for the present disclosure. Work of the presently named inventors, as well as aspects of the description that may not otherwise qualify as prior art at the time of filing, are neither expressly nor impliedly admitted as prior art.

[0004] As the world’s population grows, it is becoming increasingly important that humans produce enough food to feed all of Earth’s inhabitants. One approach to help solve the issue related to feeding the world’s population is to genetically modify plants, such as edible crops, so that such plants increase the amount of edible matter produced, are more durable, are more resilient in the face of adverse environmental or other conditions, and the like such that these crops ultimately produce more food.

[0005] One method in which genetic modification occurs is by using genetic material found in the embryonic axes of plant seeds. All plant seeds contain an embryonic axis. The genetic material of the embryonic axis can be used for genetic research and experimentation. In order to effectively access the genetic material of the embryonic axis of a plant seed, the embryonic axis is typically separated and / or extracted from the rest of the seed. The extracted embryonic axes can then be collected such that they can be manipulated for genetic research and / or experimentation.Agent Ref. No. P14771WOOO

[0006] As is well known in the art, plant tissue, particularly the interior parts of a plant seed, are somewhat fragile and susceptible to degradation or destruction when exposed to sufficient mechanized stresses or forces, to particular chemical substances, or to energy sources such as thermal or electromagnetic energy. Embryonic axes can also be relatively small and physiologically connected to other seed parts, which can make identification, access, and extraction difficult.

[0007] Existing approaches of separating / extracting embryonic axes from plant seeds involve manual extraction. Such manual extraction is performed by a researcher with basic tools such as a scalpel, forceps, and the like. Such manual extraction can only be performed one seed at a time. Thus, the process is tedious and time-consuming. Not only is the process slow, but it is costly. The slow, one-at-a-time nature of the process leads to a researcher spending large amounts of time extracting embryonic axes from seeds. Thus, if the researcher is a paid employee, contractor, and / or any other type of paid position, said researcher must be paid for that time spent extracting embryonic axes from seeds. Additionally, if a laboratory would like to increase the amount of embryonic axes extracted from seeds during a particular timespan, the laboratory would need to hire additional researchers which leads to the process being even more costly.

[0008] There are additional issues related to manual extraction. Human error is one such issue. For example, inadvertent destruction of embryonic axes can occur which wastes both the genetic material and the time spent extracting it. On the other hand, extraction and collection of embryonic axes with other seeds parts and / or debris can taint effective use of the genetic material.

[0009] Another issue with manual extraction is the ergonomic strain. Looking under a microscope for an extended period of time puts physical stress on the shoulder and back of the researcher. There is also the muscular strain on the hands of the researcher from repetitive movement.

[0010] An additional problem with manual extraction is that it requires finding specialized researchers to perform such manual extraction. Manual extraction requires fine-tune motor skills that the average person does not have. Thus, hiring, especially when scaling up an embryonic extraction process, is difficult.

[0011] Manual extraction tries to balance competing factors, including those indicated above, to non-destructively and effectively extract specific interior seed parts, including embryonic axes. But technical problems and deficiencies exist.Agent Ref. No. P14771WOOO

[0012] Thus, there exists a need in the art for an effective technological solution including an apparatus, system, and / or method which allows for automated (or at least semi-automated), rather than manual, extraction and collection of embryonic axes from plant seeds. There exists a further need in the art for an improved apparatus, system, and / or method which allows for extraction and collection of embryonic axes from plant seeds, wherein said extraction and collection is performed in a faster, more cost-effective, and less tedious manner with less risk of human error than what is known in the prior art.SUMMARY

[0013] The following objects, features, advantages, aspects, and / or embodiments, are not exhaustive and do not limit the overall disclosure. No single embodiment need provide each and every object, feature, or advantage. Any of the objects, features, advantages, aspects, and / or embodiments disclosed herein can be integrated with one another, either in full or in part.

[0014] It is a primary object, feature, and / or advantage of the present disclosure to improve on or overcome the deficiencies in the art.

[0015] It is a further object, feature, and / or advantage of aspects and / or embodiments shown and / or described in the present disclosure to provide apparatus(es), method(s), and / or system(s) configured to automatically, or at least semi-automatically, extract embryonic axes from plant seeds and collect the extracted embryonic axes in a fast, efficient, effective, and cost-effective manner.

[0016] It is still yet a further object, feature, and / or advantage of aspects and / or embodiments shown and / or described in the present disclosure to provide apparatus(es), method(s), and system(s) that are configured to rotate a suspension of whole plant seeds and a liquid in order to comminute the whole seeds and collect the extracted embryonic axes.

[0017] The apparatus(es), method(s), and system(s) disclosed herein can be used in a wide variety of applications. For example, the apparatus(es), method(s), and system(s) disclosed herein can be used for a vast array of plant seeds. The apparatus(es), method(s), and system(s) can be used for any dicot seed. Additionally, the embryonic axes extracted from seeds via the apparatus(es), method(s), and system(s) disclosed herein can be used in a variety of different ways including, but not limited to, a biolistic transformation, biolistic gene delivery, an agrobacterial transformation, and / or gene editing.

[0018] It can be beneficial that the apparatus(es), method(s), and system(s) be safe, cost- effective, and durable. For example, the apparatus(es), method(s) and system(s) disclosed herein can be configured to minimize and / or eliminate potential harm to a user. Further, theAgent Ref. No. P14771WOOO apparatus(es), method(s) and system(s) can be cost-effective, as noted above, in that they can be configured to automatically, or at least semi-automatically, extract embryonic axes from seeds in a much faster and more efficient manner than manual extraction. The apparatus(es), method(s), and system(s) greatly reduce time and effort spent extracting and collecting embryonic axes. This reduces costs associated with paying researchers and / or paying for laboratory space and / or time. Additionally, aspect(s) of the apparatus(es) and / or system(s) can be adapted to resist thermal transfer, electric conductivity, and / or failure (e.g., cracking, crumbling, shearing, creeping, and the like) due to excessive and / or prolonged exposure to tensile, compressive, and / or balanced forces acting on the apparatus(es) and / or system(s).

[0019] Methods can be practiced which facilitate use, manufacture, assembly, maintenance, and repair of the apparatus(es) and / or system(s) which accomplish some or all of the previously stated objectives.

[0020] The apparatus(es) and method(s) described herein can be incorporated into systems which accomplish some or all of the previously stated objectives. Additionally, the system(s) described herein can be incorporated into larger designs which accomplish some or all of the of the previously stated objectives. For example, the system(s) described herein can be scaled up or down to be incorporated into other designs.

[0021] According to some aspects of the present disclosure, a method for automated separation and collection of embryonic axes of dicot seeds comprises: rotating a suspension of a plurality of whole dicot seeds and a liquid relative to a surface having through-pores, the surface being effective to: comminute the whole dicot seeds into smaller pieces, and substantially pass embryonic axes; and collecting the passed embryonic axes.

[0022] According to some additional aspects of the present disclosure, the plurality of whole dicot seeds comprises a plurality of seeds of the Fabaceae family. Whole seeds of Fabaceae family members which can be used include, but are not limited to, soybean (Glycine max), peanut (Arachis hypogaea), bean (Phaseolus sp.), pea (Pisum sativum), lentil (Lens culinaris), and chickpea (Cicer arietinum) seeds.

[0023] According to some additional aspects of the present disclosure, the comminution imposes on the whole dicot seeds one or more of: friction; shear or impact stress(es); and / or degradation.

[0024] According to some additional aspects of the present disclosure, the method further comprises, while rotating the suspension, influencing the seeds toward or to the surface.Agent Ref. No. P14771WOOO

[0025] According to some additional aspects of the present disclosure, the influencing comprises one or more of: centrifugal force(s); mechanical force(s); and / or gravitational force(s).

[0026] According to some additional aspects of the present disclosure, the surface comprises: an internal portion of an inverted three-dimensional conical member positioned in an internal volume of a container, the conical member having an internal space along an axis between an open top base end and a closed opposite bottom end; and wherein the internal portion is between the open top base end and the closed opposite bottom end.

[0027] According to some additional aspects of the present disclosure, the rotating comprises rotating the suspension with an impeller: operably connected to a rotary power source; and wherein the impeller comprises an impeller body having top and bottom ends along a rotational axis, wherein the impeller body is adjustable relative to the closed opposite bottom end of the conical member.

[0028] According to some additional aspects of the present disclosure, the impeller body comprises: a structure extending towards or to the internal portion of the conical member when in operative position; bristles extending generally laterally from the rotational axis towards, at, or near the internal portion of the conical member when in operative position; or one or more flanges extending generally laterally from the rotational axis towards, at, or near the internal portion of the conical member when in operative position.

[0029] According to some additional aspects of the present disclosure, the method further comprises: autoclaving the impeller body prior to the rotation of the suspension; and sterilizing the impeller body prior to the rotation of the suspension.

[0030] According to some additional aspects of the present disclosure, the method further comprises adjusting height of the bottom end of the impeller body such that the bottom end of the impeller body is positioned within several inches from the closed opposite bottom end of the conical member.

[0031] According to some additional aspects of the present disclosure, the method further comprises securing the conical member to the container via a clamp prior to the rotating of the suspension.

[0032] According to some additional aspects of the present disclosure, the collecting of the passed embryonic axes further comprises at least one of the following steps: removing the conical member from the internal volume of the container and pouring out excess liquid from the container wherein at least a portion of debris is poured out with the excess liquid but the passed embryonic axes generally remain in the container; adding an additional amount of liquidAgent Ref. No. P14771WOOO to the container; moving a suspension remaining in the container wherein said remaining suspension comprises at least a portion of the liquid, the additional amount of liquid, the passed embryonic axes, and another portion of the debris; pouring the remaining suspension into a second container; pouring out a majority of liquid from the second container; moving the second container to separate the passed embryonic axes in the second container from debris in the second container; pouring out the debris from the second container while the passed embryonic axes remain in the second container; resuspending the passed embryonic axes in the second container; and / or pouring the passed embryonic axes into a third container.

[0033] According to some additional aspects of the present disclosure, the additional amount of liquid is 500 milliliters.

[0034] According to some additional aspects of the present disclosure, the third container is a petri dish.

[0035] According to some additional aspects of the present disclosure, the method further comprises suspending the passed embryonic axes in a tissue culture medium (e.g., an Agrobacterium co-culture medium or biolistic pre-bombardment medium).

[0036] According to some additional aspects of the present disclosure, the plurality of whole dicot seeds are soybean seeds; a pore size of each of the through-pores is on the order of or 6 millimeters; and the suspension is rotated on the order of 75 to 200 or 300 RPM.

[0037] According to some additional aspects of the present disclosure, the collecting of the passed embryonic axes is for the purpose of one or more of: a biolistic transformation; biolistic gene delivery; an agrobacterial transformation; and / or gene editing.

[0038] According to some additional aspects of the present disclosure, the whole dicot seeds are imbibed for several hours prior to being rotated in the suspension.

[0039] According to some additional aspects of the present disclosure, the liquid is sterile water.

[0040] According to some other aspects of the present disclosure, an apparatus for automated separation and collection of embryonic axes of dicot seeds comprises: a container having: an internal volume configured to receive and retain a suspension of a set of imbibed whole dicot seeds and a liquid; an inverted three-dimensional conical member positioned at least partially in the internal volume of the container, the conical member having: an internal space along an axis between an open top base end and a closed opposite bottom end; and an internal surface between the open top base end and the closed opposite bottom end with through-pores, the internal surface being effective to: comminute the imbibed whole dicot seeds; and substantially pass embryonic axes of each of the imbibed whole dicot seeds; an impeller having: an interface configured to operably connect to a rotary power source; and an impeller body extending from aAgent Ref. No. P14771WOOO base end to an opposite end along a rotational axis, the impeller body configured to: rotate the suspension; and influence the set of imbibed whole dicot seeds toward or to the internal surface of the conical member; and a collection area in the container outside the internal space of the conical member to collect embryonic axes that have passed through the through-pores.

[0041] According to some additional aspects of the present disclosure, the conical member is a circular conical member.

[0042] According to some additional aspects of the present disclosure, the conical member is a truncated cone with the closed opposite bottom end closed at a truncation plane.

[0043] According to some additional aspects of the present disclosure, the internal surface and the impeller together are configured to degrade and / or comminute the set of imbibed whole dicot seeds.

[0044] According to some additional aspects of the present disclosure, a pore size of each of the through-pores is on the order of or 6 millimeters.

[0045] According to some additional aspects of the present disclosure, the impeller body has a cylindrical shape, conical shape, substantially cylindrical shape, or substantially conical shape.

[0046] According to some additional aspects of the present disclosure, the impeller body comprises bristles extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.

[0047] According to some additional aspects of the present disclosure, at least some of the bristles extend to the internal surface of the conical member when in operative position.

[0048] According to some additional aspects of the present disclosure, the impeller body comprises a tire brush.

[0049] According to some additional aspects of the present disclosure, the impeller body comprises one or more flanges extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.

[0050] According to some additional aspects of the present disclosure, the liquid is sterile water.

[0051] According to some additional aspects of the present disclosure, the apparatus further comprises a clamp to attach the conical member to the container such that the axis of the conical member is oblique to the rotational axis of the impeller.

[0052] According to some additional aspects of the present disclosure, the apparatus further comprises a stand configured to secure the rotary power source and the container in place during operation, the stand comprising: a base; a base securing member; an elongated member; and an upper member.Agent Ref. No. P14771WOOO

[0053] According to some additional aspects of the present disclosure, the apparatus in combination with a second container is used for collection of the collected embryonic axes that have passed through the through-pores.

[0054] According to some additional aspects of the present disclosure, the container comprises: an internal volume of on the order of 10 L; a height on the order of 12 inches; and a diameter on the order of 10.5 inches.

[0055] According to some other aspects of the present disclosure, a system for automated separation and collection of embryonic axes of dicot seeds comprises: means for rotating a suspension comprising a set of imbibed whole dicot seeds and a liquid relative to a surface having through-pores, the surface being effective to: comminute the set of imbibed whole dicot seeds into smaller pieces; and substantially pass embryonic axes; and means for collecting passed embryonic axes.

[0056] According to some additional aspects of the present disclosure, the surface comprises: an internal portion of an inverted three-dimensional conical member positioned in an internal volume of a container, the conical member having an internal space along an axis between an open top base end and a closed opposite bottom end; and wherein the internal portion is between the open top base end and the closed opposite bottom end.

[0057] According to some additional aspects of the present disclosure, the system further comprises an impeller having: an interface operably connected to a controllable rotary power source; and an impeller body extending from a base end to an opposite end along a rotational axis, the impeller body configured to: rotate the suspension; and influence the set of imbibed whole dicot seeds toward or to the internal portion of the conical member.

[0058] According to some additional aspects of the present disclosure, the means for collecting comprises the container.

[0059] According to some additional aspects of the present disclosure, the means for collecting further comprises a second container, the second container configured to receive the passed embryonic axes from the first container.

[0060] According to some additional aspects of the present disclosure, the system further comprises a clamp to releasably attach the conical member to the container such that the axis of the conical member is oblique to the rotational axis of the impeller.

[0061] According to some additional aspects of the present disclosure, the system further comprises a stand configured to secure the rotary power source and the container in place during operation, the stand comprising: a base; a base securing member; an elongated member; and an upper member.Agent Ref. No. P14771WOOO

[0062] According to some additional aspects of the present disclosure, the conical member is circular.

[0063] According to some additional aspects of the present disclosure, the conical member is a truncated cone with the closed opposite bottom end closed at a truncation plane.

[0064] According to some additional aspects of the present disclosure, the container comprises: an internal volume of on the order of 10 L; a height on the order of 12 inches; and a diameter on the order of 10.5 inches.

[0065] According to some additional aspects of the present disclosure, the internal portion of the conical member and the impeller together are configured to degrade and / or comminute the set of imbibed whole dicot seeds.

[0066] According to some additional aspects of the present disclosure, the impeller body has a cylindrical shape, conical shape, substantially cylindrical shape, or substantially conical shape.

[0067] According to some additional aspects of the present disclosure, the impeller body comprises one or more flanges extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.

[0068] According to some additional aspects of the present disclosure, the impeller body comprises bristles extending generally laterally from the rotational axis towards the internal portion of the conical member when in operative position.

[0069] According to some additional aspects of the present disclosure, the bristles extend to the internal portion of the conical member when in operative position.

[0070] According to some additional aspects of the present disclosure, the impeller body comprises a tire brush.

[0071] According to some additional aspects of the present disclosure, a pore size of each of the through-pores is on the order of or 6 millimeters.

[0072] According to some additional aspects of the present disclosure, the liquid is sterile water.

[0073] These and / or other objects, features, advantages, aspects, and / or embodiments will become apparent to those skilled in the art after reviewing the following brief and detailed descriptions of the drawings. Furthermore, the present disclosure encompasses aspects and / or embodiments not expressly disclosed but which can be understood from a reading of the present disclosure, including at least: (a) combinations of disclosed aspects and / or embodiments and / or (b) reasonable modifications not shown or described.Agent Ref. No. P14771WOOOBRIEF DESCRIPTION OF THE DRAWINGS

[0074] Several embodiments in which the present disclosure can be practiced are illustrated and described in detail, wherein like reference characters represent like components throughout the several views. The drawings are presented for exemplary purposes and may not be to scale unless otherwise indicated.

[0075] In addition, as will be understood, any of the aspects of any of the embodiments shown and / or described herein could be combined with one another to form any number of embodiments, whether expressly disclosed or not, which would be understood by one skilled in the art.

[0076] Figure 1A shows a side elevation transparent perspective diagrammatic view of an embryonic axis extraction and collection apparatus according to at least some aspects of the present disclosure.

[0077] Figure IB shows a side elevation transparent perspective view of an example embodiment of aspects of a conical member and impeller according to at least some aspects of the present disclosure.

[0078] Figure 1C shows a side elevation transparent perspective view of another example embodiment of aspects of a conical member and impeller according to at least some aspects of the present disclosure.

[0079] Figure ID shows a side elevation transparent perspective view of another example embodiment of aspects of a conical member and impeller according to at least some aspects of the present disclosure.

[0080] Figure IE shows a side elevation transparent perspective view of another example embodiment of aspects of a conical member and impeller according to at least some aspects of the present disclosure.

[0081] Figure IF shows a side elevation transparent perspective view of another example embodiment of aspects of a conical member and impeller according to at least some aspects of the present disclosure.

[0082] Figure 2 shows a perspective view of a 2000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the 2000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art according to at least some aspects of the present disclosure.

[0083] Figure 3 shows a perspective view of a 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art, wherein aspect(s) of the embryonic axis extraction andAgent Ref. No. P14771WOOO collection apparatus of Figure 1A can comprise the 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art according to at least some aspects of the present disclosure.

[0084] Figure 4 shows a perspective view of a Crofton Cone Grater produced by Aldi, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the Crofton Cone Grater produced by Aldi according to at least some aspects of the present disclosure.

[0085] Figure 5 shows a Lab Digital Overhead Stirrer Lab Mixer manufactured by XZBELEC, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the Lab Digital Overhead Stirrer Lab Mixer manufactured by XZBELEC according to at least some aspects of the present invention.

[0086] Figures 6A-F show various views and / or aspects of a 60L Overhead Stirrer produced by +UXI, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the 60L Overhead Stirrer produced by +UXI according to at least some aspects of the present disclosure.

[0087] Figures 7A-C shows various views and / or aspects of a Bottle Brush Drill Accessory produced by RotoScrub, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the Bottle Brush Drill Accessory produced by RotoScrub according to at least some aspects of the present disclosure.

[0088] Figure 8 shows a perspective view of a Paint Mixer Drill Attachment produced by Edward Tools, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the Paint Mixer Drill Attachment produced by Edward Tools according to at least some aspects of the present disclosure.

[0089] Figure 9 shows a perspective view of a 43420 Paint Mixer produced by Hyde, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the 43420 Paint Mixer produced by Hyde according to at least some aspects of the present disclosure.

[0090] Figure 10 shows a perspective view of a Drill Brush Power Scrubber by Useful Products - Drillbrush Soft White Automotive Cleaning kit with Extended Reach Attachment - Drill Bit Extension Carpet Cleaner Solution - Car Interior Brush Set, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 1A can comprise the Drill Brush Power Scrubber by Useful Products according to at least some aspects of the present disclosure.

[0091] Figure 11 shows a perspective view of a media bottle according to at least some aspects of the present disclosure.Agent Ref. No. P14771WOOO

[0092] Figure 12 shows a perspective view of an embryonic axis extraction and collection apparatus according to at least some aspects of the present disclosure.

[0093] Figure 13 shows another perspective view of some components of the embryonic axis extraction and collection apparatus of Figure 12.

[0094] Figure 14 shows an enlarged perspective view of some components of the embryonic axis extraction and collection apparatus of Figure 12.

[0095] Figure 15 shows an isolated perspective view of a stopper that can be used with the embryonic axis extraction and collection apparatus of Figure 12.

[0096] Figure 16 shows a top perspective view of some components that can be used with the embryonic axis extraction and collection apparatus of Figure 12.

[0097] Figure 17 shows a side elevation perspective view of some components of the embryonic axis extraction and collection apparatus of Figure 12.

[0098] Figure 18 shows a perspective view of a Multi-Lens LED Magnifier manufactured by ULINE, wherein aspect(s) of the embryonic axis extraction and collection apparatus of Figure 12 can comprise the Multi-Lens LED Magnifier manufactured by ULINE according to at least some aspects of the present disclosure.

[0099] Figure 19A shows a perspective view of a partially assembled example embodiment of an embryonic axis extraction and collection apparatus according to at least some aspects of the present disclosure.

[0100] Figure 19B shows a perspective view of portion(s) of the conical member of Figure 19A wherein example dimensions of a closed bottom end of the conical member are shown.

[0101] Figure 19C shows a side perspective view of the conical member of Figure 19A wherein example dimensions of an open top base end of the conical member are shown.

[0102] Figure 19D shows a side perspective view of the conical member of Figure 19A wherein example dimensions of the diagonal length of the conical member are shown.

[0103] Figure 19E shows an enlarged side elevation view of one through-pore of the conical member of Figure 19A, wherein example dimensions of the through-pore and / or a dome thereof are shown.

[0104] Figure 19F shows an enlarged side elevation view of one through-pore of the conical member of Figure 19A, wherein example dimensions of the dome thereof are shown.

[0105] Figure 19G shows an enlarged side perspective view of aspect(s) of the conical member of Figure 19A, wherein example dimensions of a dome are shown.Agent Ref. No. P14771WOOO

[0106] Figure 20 shows a perspective view of a partial assembly of an example embodiment of an embryonic axis extraction and collection apparatus, according to at least some aspects of the present disclosure.

[0107] Figure 21 shows a perspective view of the impeller of the embryonic axis extraction and collection apparatus of Figure 20 in isolation.

[0108] Figure 22 shows a perspective view of the container of the embryonic axis extraction and collection apparatus of Figure 20 wherein the container includes a cover.

[0109] Figure 23 shows a top elevation view of the container of the embryonic axis extraction and collection apparatus of Figure 20 wherein the container includes the cover.

[0110] Figure 24 shows a perspective view of the container of the embryonic axis extraction and collection apparatus of Figure 20 wherein the container includes a lip cover to cover the spout.

[0111] Figure 25 shows a perspective view of the impeller of the embryonic axis extraction and collection apparatus of Figure 20 and the Paint Mixer Drill Attachment produced by Edward Tools of Figure 8, both in isolation.

[0112] Figure 26 shows a greatly enlarged view of a dicot seed sectioned into halves, illustrating an exposed embryonic axis inside the seed.

[0113] Figure 27 shows an example method of embryonic axis extraction and collection according to at least some aspects of the present disclosure.

[0114] Figure 28 shows another example method of embryonic axis extraction and collection according to at least some aspects of the present disclosure.

[0115] Figures 29A-E show an example method of embryonic axis extraction and collection according to at least some aspects of the present disclosure.

[0116] Figure 30 shows a diagram of a system including an embryonic axis extraction and collection apparatus according to at least some aspects of the present disclosure.

[0117] Figure 31 shows an isolated perspective view of aspects of the system of Figure 30.

[0118] Figure 32A shows a front elevation view of a Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7, wherein aspect(s) of the system of Figure 30 can comprise the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 according to at least some aspects of the present disclosure.

[0119] Figure 32B shows a side perspective view of the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 of Figure 32A.

[0120] Figure 32C shows a bottom perspective view of the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 of Figure 32A.Agent Ref. No. P14771WOOO

[0121] Figure 33 shows a perspective view of a Polypropylene Beaker with Handle manufactured by Carolina Biological Supply, wherein aspect(s) of the system of Figure 30 can comprise the Polypropylene Beaker with Handle manufactured by Carolina Biological Supply according to at least some aspects of the present disclosure.

[0122] Figure 34 shows a perspective view of a Nalgene Autoclavable Polypropylene Pan manufactured by Thermo Scientific, wherein aspect(s) of the system of Figure 30 can comprise the Nalgene Autoclavable Polypropylene Pan manufactured by Thermo Scientific according to at least some aspects of the present disclosure.

[0123] An artisan of ordinary skill in the art need not view, within isolated figure(s), the near infinite number of distinct permutations of features described in the following detailed description to facilitate an understanding of the present disclosure.DETAILED DESCRIPTION

[0124] The present disclosure is not to be limited to that described herein. Mechanical, electrical, chemical, procedural, and / or other changes can be made without departing from the spirit and scope of the present disclosure. No features shown or described are essential to permit basic operation of the present disclosure unless otherwise indicated.

[0125] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the present disclosure pertain.

[0126] The terms “a,” “an,” and “the” include both singular and plural referents.

[0127] The term “or” is synonymous with “and / or” and means any one member or combination of members of a particular list.

[0128] The terms “invention” or “present invention” are not intended to refer to any single embodiment of the particular invention but encompass all possible embodiments as described in the specification and the claims.

[0129] The term “about” as used herein refers to slight variations in numerical quantities with respect to any quantifiable variable. Inadvertent error can occur, for example, through use of typical measuring techniques or equipment or from differences in the manufacture, source, or purity of components.

[0130] The term “substantially” refers to a great or significant extent. “Substantially” can thus refer to a plurality, majority, and / or a supermajority of said quantifiable variable, given proper context.

[0131] The term “generally” encompasses both “about” and “substantially.”Agent Ref. No. P14771WOOO

[0132] The term “configured” describes structure capable of performing a task or adopting a particular configuration. The term “configured” can be used interchangeably with other similar phrases, such as “constructed”, “arranged”, “adapted”, “manufactured”, and the like.

[0133] Terms characterizing sequential order, a position, and / or an orientation are not limiting and are only referenced according to the views presented.

[0134] The “scope” of the present disclosure is defined by the appended claims, along with the full scope of equivalents to which such claims are entitled. The scope of the disclosure is further qualified as including any possible modification to any of the aspects and / or embodiments disclosed herein which would result in other embodiments, combinations, subcombinations, or the like that would be obvious to those skilled in the art.

[0135] As used herein, the term “exemplary” refers to an example, an instance, or an illustration, and does not indicate a most preferred embodiment unless otherwise stated.

[0136] Referring now to the figures, an embryonic axis extraction and collection apparatus 10 is shown in Figure 1A. The embryonic axis extraction and collection apparatus 10 is configured to separate and collect embryonic axes of whole dicot seeds 18, shown diagrammatically as squares in Figure 1A, in an automated fashion, or at least a semi-automated fashion. As shown in Figure 1A, according to at least some embodiments, the embryonic axis extraction and collection apparatus 10 can include a container 12, a conical member 22, a rotary power source 34, and an impeller 36.

[0137] The container 12 can be any sort of receptacle capable of holding a liquid including any number of commercially available off-the-shelf receptacles. For example, the container 12 could be any sort of beaker, pitcher, cup, bucket, and the like that is configured to receive and retain a suspension of whole dicot seeds and a liquid. The container can comprise a central longitudinal axis 13. According to some embodiments, the container 12 can be the 2000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12A (UNSPSC code: 41121803; catalog number F28992-0000; UPC number 191634064527) (see www.mscdirect.com / product / detail s / 65362576?cid=ppc-google- &mkwid=%7Cdc&pcrid=&rd=k&product_id=65362576&gad_source=l&gclid=Cj0KCQjw3Za yBhDRARIsAPWzx8qN4MgA3VybpeXe- dU6rI3vQnDwDs2JBwrbMDifgqBfgOEfrSVmRv4aAmOXEALw_wcB&gclsrc=aw.ds (date accessed: October 18, 2024)) (see also www.belart.eom / bel-art-f28992-0000-tall-form-2000ml- polypropylene-graduated-pitcher-20ml-graduation.html (date accessed: October 18, 2024)). An example image of the 2000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12A is shown in Figure 2. According to some embodiments, the 2000 milliliter PolypropyleneAgent Ref. No. P14771WOOOGraduated Pitcher manufactured by Bel-Art 12A can include a handle, can be sterile, can be autoclavable, can be translucent, can be configured to be able to be used in temperatures up to about 250 degrees Fahrenheit (121 degrees Celsius) and / or to be able to hold material (such as liquid material) that is up to about 250 degrees Fahrenheit (121 degrees Celsius), can be graduated every 20 milliliters, can have a volume of about 2000 milliliters, and can have the following dimensions: a length / height of about 8.5 inches (21.5 centimeters), a top diameter of about 5 and 7 / 8 inches (15 centimeters), and a bottom diameter of about 4.5 inches (11.6 centimeters).

[0138] According to some embodiments, the container 12 can be the SP Bel-Art Tall Form 10,000 milliliter polypropylene graduated pitcher 12B (Catalog No. : F28995-0000; UPC No. : 191634013761) (see www.belart.eom / bel-art-f28995-0000-tall-form- 10-000ml-polypropylene- graduated-pitcher-200ml-graduation.html (date accessed: October 18, 2024)). The 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12B can include 200 milliliter graduation. The 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12B can comprise a flared rim(s) and spout(s) for quick, easy pouring when large volumes are handled. The 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12B can provide for good chemical resistance. The 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel- Art 12B can be steam autoclavable at 121 degrees Celsius (250 degrees Fahrenheit). An example image of the 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12B is shown in Figure 3. According to some embodiments, the 10,000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12B can be graduated every 200 milliliters, can have a volume of about 10,000 milliliters, and can have the following dimensions: a height of about 12 and 5 / 8 inches (32.0 centimeters), a top diameter of about 10 and 5 / 8 inches (26.9 centimeters), and a bottom diameter of about 8 and % inches (22.3 centimeters).

[0139] According to some embodiments, the container 12 can have a volume of about 2 liters, a height of about 8 inches, and a diameter of about 4.5 inches. According to some embodiments, the container 12 can have a volume of about 0.9985 liters, a height of about 7 inches, and a diameter of about 4.5 inches. According to various embodiments, the container 12 can have an internal volume 14 of about 1 liter to about 2 liters, a height of about 7 inches to about 8 inches, and a diameter of about 4 inches to about 5 inches. According to some embodiments, the container 12 can have an internal volume 14 of approximately 10 liters, a height of approximately 12 inches, and a diameter of approximately 10.5 inches. However, the container 12 can include any suitable dimensions according to need and / or desire. The container 12 isAgent Ref. No. P14771WOOO configured to include an internal volume 14 and a collection area 16 wherein extracted embryonic axes can be collected. The container 12 can be hollow and can be generally cylindrical, according to some embodiments, wherein the container 12 is enclosed on the sides and the bottom with an open top, and wherein the hollow nature of the container 12 creates the internal volume 14. The collection area 16 of the container 12 can generally be located at and / or near the bottom of the container 12. According to some embodiments, the collection area 16 is the closed end and / or floor of the container 12. Substantially extracted embryonic axes can be collected at and / or near the collection area 16.

[0140] When the apparatus 10 is in use, the conical member 22 is configured to be positioned, at least partially, within the internal volume 14 of the container 12 as shown in Figure 1A. The conical member 22 can be three-dimensional and can be inverted such that the open top base end 28 faces upward and the closed opposite bottom end 30 faces downward. The conical member 22 in this example can be any sort of conical structure having at least one aperture and / or pore through which liquids and solids smaller than the aperture and / or pore can pass. For example, the conical member 22 can be any sort of conical grater having at least one aperture and / or pore. According to some embodiments, the conical member 22 can comprise and / or substantially be made of stainless steel. According to some embodiments, the conical member 22 can be the Crofton Cone Grater produced by Aldi 22 A (see www.instacart.com / store / aldi / products / 32398465?guest=true (date accessed: October 18, 2024)). An example image of the Crofton Cone Grater produced by Aldi 22A is shown in Figure 4. According to some embodiments, the Crofton Cone Grater produced by Aldi 22A can be a cone grater, rotary grater, or microplane grater; can include 3 different grating surfaces including course, fine, and extra fine grating blades; can include a stainless steel grating cylinder and a soft grip handle and knob; and / or can include dual size blades and a soft grip handle.

[0141] While the conical member of some embodiments, such as that of Figure 1 A, include one or more through-pores generally positioned only on one portion of the surface of the conical member (such as one-third of the surface of the conical member), according to some embodiments, such as that of Figures 19A-G, one or more through-pores can be positioned on all sides of the surface of (i.e., all around) the conical member. Further, while the conical member of some embodiments may have through-pores of differing sizes, the conical member of other embodiments, such as that of Figures 19A-G, can have through-pores of substantially uniform size. Having through-pores of substantially uniform size wherein the through-pores are positioned on all sides of the surface of the conical member can allow for better and / or more efficient extraction than when through-pores are not positioned on all sides of the surface of theAgent Ref. No. P14771WOOO conical member, such as when they are positioned only on one-third of the surface of the conical member, and / or when the through-pores have differing sizes.

[0142] According to some embodiments, the conical member 22 could be a strainer rather than a grater. According to some embodiments, the conical member 22 comprises a circular conical shape. According to some embodiments, the conical member 22 is configured to include an internal space 24, an internal surface / portion 26, an open top base end 28, a closed opposite bottom end 30, and at least one through-pore 32.

[0143] The conical member 22 can be hollow, wherein the internal space 24 is located in the hollow portion of the conical member 22. The internal space 24 can be positioned along a central longitudinal axis 23 of the conical member 22 that extends from the open top base end 28 to the closed opposite bottom end 30. The internal surface 26 of the conical member 22 is the inner surface of the side, sloped portions of the conical member 22 positioned between the open top base end 28 and the closed opposite bottom end 30. The internal surface 26 surrounds the internal space 24 of the conical member 22. The internal surface 26 comprises at least one through-pore 32. While the conical member 22 of the example embodiment shown in Figure 1A diagrammatically illustrates six through-pores 32 with circle symbols, according to various embodiments, the conical member 22 can include any number of through-pores ranging from 1 to N where N is any number greater than one. Additionally, while the example embodiment of Figure 1A diagrammatically illustrates the through-pores 32 arranged in a generally straight line, the through-pores 32 could be arranged in any suitable configuration on the conical member 22 such as in a circular configuration, in any sort of patterned configuration, in a random configuration, and / or in any other suitable configuration.

[0144] The open top base end 28 of the conical member 22 is positioned at the end of the conical member 22 that has the larger diameter. The open top base end 28 is configured to be open such that materials, such as solid and / or liquid materials, can enter the conical member 22 via the open top base end 28 wherein said materials have access to the internal space 24 and the internal surface 26 of the conical member 22. The conical member 22 is configured to be positioned such that the open top base end 28 is directed upward.

[0145] The closed opposite bottom end 30 is located at the opposite end of the conical member 22 as the open top base end 28. The closed opposite bottom end 30 is located at the end of the conical member having a smaller diameter than that of the other end. The closed opposite bottom end 30 is closed such that no material can enter and / or exit the internal space 24 of the conical member 22 via the closed opposite bottom end 30. The closed opposite bottom end 30 can be naturally closed, such as an original apex of a cone, such that the internal surface 26 ofAgent Ref. No. P14771WOOO the conical member 22 encapsulates the closed opposite bottom end 30. Alternatively, according to some embodiments, the closed opposite bottom end 30 can be naturally open, such as an originally truncated cone, such that the materials, such as solid or liquid material, could enter and exit the internal space 24 of the conical member 22 via that originally open end. In such embodiments wherein the closed opposite bottom end 30 is naturally open, a stopper (not shown in Figure 1A) (or any other suitable closing member capable of closing the truncated opposite bottom end 30 of the conical member 22) can be inserted at and / or into the closed opposite bottom end 30 to effectively close the closed opposite bottom end 30 such that no materials, such as solid or liquid materials, can enter and / or exit the internal space 24 of the conical member 22 via the closed opposite bottom end 30. The stopper can be any sort of stopper including, but not limited to, a rubber stopper, a plastic stopper, and the like. Alternatively, the closed opposite bottom end 30 of the conical member 22 can comprise a true conical closed apex, rather than a truncated apex, wherein a stopper, or other closing member, is unnecessary. According to embodiments that include the stopper, the conical member 22 can be truncated such that the stopper effectively closes the closed opposite bottom end 30 at the truncation plane. As noted, according to some embodiments such as that shown in Figures 19A-G, the closed opposite bottom end of the conical member can be truncated but can also be originally closed such that no stopper, or any other type of closing member, is necessary to close the closed opposite bottom end.

[0146] As noted, the internal surface 26 of the conical member 22 includes at least one through- pore 32. The at least one through-pores 32 can have a pore size wherein said pore size is equal to the diameter of the at least one through-pores 32. The at least one through-pores 32 can have a pore size effective to substantially pass substantially intact embryonic axes. According to at least one non-limiting example, the pore size of the at least one through-pores 32 is about 6 millimeters. Alternatively, the pore size of the at least one through-pores 32 can be any suitable size capable of stopping a whole dicot seed while also capable of substantially passing the substantially intact embryonic axis of each seed. Thus, the size of the at least one through-pores 32 can vary depending on the type of seed whose embryonic axis is being extracted and collected. According to some embodiments, the pore size of the at least one through-pores 32 can range from about 5 millimeters to about 6 millimeters. According to some embodiments, the conical member 22 can comprise through-pores 32 wherein the through-pores 32 have different sizes. For example, according to some embodiments, the conical member 22 can comprise through-pores 32 wherein each through-pore 32 is one of three different sizes. In other words, according to some embodiments, the conical member 22 could comprise one or more through-Agent Ref. No. P14771WOOO pores 32 having a first pore size, one or more through-pores 32 having a second pore size, and one or more through-pores 32 having a third pore size. As noted, according to some embodiments, the size of each of the one or more through-pores 32 can be substantially the same. According to some embodiments, it can be beneficial to include one or more through- pores 32 having substantially the same size as different sized through-pores can negatively affect the efficiency of extraction under particular circumstances. It should be noted that only one of the through-pores 32 shown in Figure 1A is labeled. There can be benefits to having greater than 1 through-pores. One such benefit is higher through put per unit time during the separation and / or extraction operation of the apparatus 10.

[0147] According to some embodiments, an outer surface of the conical member 22 can include one or more raised domes at least partially covering each through-pore 32. Each dome can be configured to protrude outwardly from the outer surface of the conical member 22 such that each dome partially covers at least a portion of each through-pore 32. According to some embodiments, each through-pore 32 can comprise a raised dome such that the number of through-pores 32 is equal to the number of raised domes. Each dome can have a top and a bottom. The top of each dome can be operatively attached and / or in contact with the outer surface of the conical member 22, and the bottom of each dome can be generally open and / or unattached to the conical member 22 such that any materials (such as liquid and / or embryonic axes) that pass through the through-pores 32 can exit the internal space 24 of the conical member 22 and collect in the container 12. The top of each dome can be generally circular, spherical, and / or ovate in shape, however, any suitable shape could be used. The bottom of each dome could be generally straight such that the bottom comprises a cut-off and / or truncation line of a truncated sphere and / or ovoid shape. However, any suitable shape and / or configuration could be used. When the conical member 22 is in the operative, inverted position such that the open top base end 28 faces upward (i.e., positioned as the “top” end) and the closed opposite bottom end 30 faces downward (i.e., positioned as the “bottom” end) each dome can protrude on the outer surface of the conical member 22 at or near the top of each through-pore 32 and extend downward such that a top portion of each through-pore 32 is at least partially covered. The bottom of each through-pore 32 can be substantially uncovered. Each dome can be substantially circular, spherical, and / or ovate in shape. For example, according to some embodiments, each dome can comprise one quarter, and / or similar to one quarter, of a complete sphere and / or ovoid. According to some embodiments, the height of each dome, meaning the distance each dome extends from the outer surface of the conical member 22, can be about 1 to 2 millimeters. However, according to various embodiments, the height of each dome can vary based on factorsAgent Ref. No. P14771WOOO including, but not limited to, the size of the seed and / or genotype, the size of the through-pores 32, and the like.

[0148] The rotary power source 34 can be any sort of motor and / or rotator capable of rotating an impeller. For example, the rotary power source 34 can be any sort of motor, engine, machine, and the like capable of imparting rotational movement to an impeller. According to some embodiments, the rotary power source 34 can be and / or comprise the Lab Digital Overhead Stirrer Lab Mixer manufactured by XZBELEC 34A (item number 8856587459593) (see www.amazon. com / Overhead-Lab oratory -industrial -O-lOOOOmpas- Warranty / dp / B07C7FNNH3 / ref=asc_df_B07C7FNNH3 / ?tag=hyprod- 20&linkCode=df0&hvadid=312193624974&hvpos=&hvnetw=g&hvrand= 14597012902061643 896&hvpone=&hvptwo=&hvqmt=&hvdev=c&hvdvcmdl=&hvlocint=&hvlocphy=9016722&hv targid=pla-568831710362&psc=l&mcid=b7bb998575af3e569da00ae632286efd&tag=&ref=&adgrpid=615 97290225&hvpone=&hvptwo=&hvadid=312193624974&hvpos=&hvnetw=g&hvrand=145970 12902061643896&hvqmt=&hvdev=c&hvdvcmdl=&hvlocint=&hvlocphy=9016722&hvtargid= pla-568831710362 (date accessed: October 18, 2024)). An example image of the Lab Digital Overhead Stirrer Lab Mixer manufactured by XZBELEC 34A is shown in Figure 5. According to some embodiments, the Lab Digital Overhead Stirrer Lab Mixer manufactured by XZBELEC 34A can have a speed range of about 60-2000 rpm, a motor rating input voltage of about 110 V (60 Hz), about a 20 liter capacity, a torque maximum of about 1850 gem, a chuck range minimum of about 0.5 millimeter, a chuck range maximum of about 10 millimeters, a permissible ambient temperature of about 5 to 40 degrees Celsius, a permissible relative moisture rating of about 80%, can be usable for material with a viscosity of 0-10,000 mPas, and / or can comprise a quiet, brushless motor wherein said motor has a 120 W speed range.

[0149] According to some embodiments, the rotary power source 34 can be and / or comprise the 60L Overhead Stirrer produced by +UXI 34B (ASIN: B07W49155L) (see www.amazon. com / dp / B07W49155L / ref=sspa_dk_detail_l?pd_rd_i=B07W49155L&pd_rd_w= M96v4&content-id=amzn 1. sym.7446a9d 1 -25fe-4460-b 135-a60336bad2c9&pf_rd_p=7446a9d 1 - 25fe-4460-bl35- a60336bad2c9&pf_rd_r=P 1 VPG75D5X3320G10WFW&pd_rd_wg= V49Wf&pd_rd_r=6dda9ac 6-7aa8-4950-9251- 360f36d9ccb5&s=industrial&sp_csd=d21kZ2V0TmFtZTlzcF9kZXRhaWw&th=l (date accessed: October 18, 2024)). Example images of the 60L Overhead Stirrer manufactured by +UXI 34B, and / or components thereof, are shown in Figures 6A-F. While it is said above thatAgent Ref. No. P14771WOOO the Overhead Stirrer produced by +UXI 34B is sized for 60 liters, the Overhead Stirrer produced by +UXI 34B could be sized for any suitable volume such as 20 liters, 40 liters, 60 liters, 100 liters, and the like. According to some embodiments, the Overhead Stirrer produced by +UXI 34B can have dimensions of about 31.5 inches (height) x 15.7 inches (length) x 5.5 inches (width) and can weigh about 25.8 pounds inclusive of a stand and base. According to some embodiments, the Overhead Stirrer produced by +UXI 34B can have dimensions of about 30.7 inches (height) x 12.4 inches (length) x 7.87 inches (width) inclusive of a stand and base. According to some embodiments, when not mounted on a stand, the Overhead Stirrer produced by +UXI 34B can have dimensions of about 6.69 inches (height) x 3.07 inches (width) x 1.77 inches (diameter of portion that connects to impeller). According to some embodiments, when not mounted on a stand, the Overhead Stirrer produced by +UXI 34B can have exterior dimensions of about 186 mm x 83 mm x 220 mm. According to some embodiments, the Overhead Stirrer produced by +UXI 34B can be attached to an impeller interface having a diameter of 0.5 to 10 millimeters. According to some embodiments, the Overhead Stirrer produced by +UXI 34B can have a speed range of about 30-2200 rpm, a timer range of 1-9999 minutes, a speed resolution of about plus or minus 1 rpm, a torque rating of 40, 60, or 80 N*cm, can be configured for use with a material (such as a liquid material) having a maximum viscosity of about 80,000 mPas, can be configured for use in an ambient temperature of about 5- 40 degrees Celsius, can be configured for an input power of about 60 to 160 W, can be configured for an output power of about 50 to 150 W, can be configured for a power rating of about 70 to 160 W, and / or can be configured for use when ambient moisture is about 80%. The Overhead Stirrer produced by +UXI 34B can include a safety circuit, motor protection, and / or overload protection, such as multiple overload protection. The Overhead Stirrer produced by +UXI 34B can include an LCD display to display speed of rotation and / or time. The Overhead Stirrer produced by +UXI 34B can be used with multiple different types of impellers. The portion of the Overhead Stirrer produced by +UXI 34B that is configured to connect to an impeller interface can include a silicon cover for protection and / or can include a self-locking drill clip that is easy to fasten and can be used without tools. The stand of the Overhead Stirrer produced by +UXI 34B can include shedding protection, a non-slip mat / base for stability, and a chuck having an adjustable height which can be adjusted without a tool. The Overhead Stirrer produced by +UXI 34B can include a push-up rod that makes it easy to assemble and / or disassemble the Overhead Stirrer produced by +UXI 34B in case of a need to replace it. The Overhead Stirrer produced by +UXI 34B can include automatic torque adjustment, speed and time control, low noise, and / or a brushless DC motor.Agent Ref. No. P14771WOOO

[0150] According to some embodiments, the rotary power source 34 can be and / or comprise an electric motor powered by plugging into the normal electric grid wherein power is supplied as 110 VAC, 120 VAC, and / or any other suitable power level. According to some embodiments, the rotary power source 34 could be battery-powered. The rotary power source 34 can be operatively attached to the impeller 36 such that the rotary power source 34 rotates the impeller 36 and / or causes the impeller 36 to rotate along a rotational axis 45. According to some embodiments, the rotary power source 34 can be and / or comprise magnetic steering in addition to or alternative to a motor. In Figure 1A, since, according to some embodiments, the longitudinal axis 13 of the container 12, the longitudinal axis 23 of the conical member 22, and the rotational axis 45 of the impeller 36 are aligned, each of the axes 12, 23, and 45 are shown as a single axis. However, as noted herein, according to various embodiments, any combination of the axes 12, 23, 45 can be aligned and / or offset with each other.

[0151] As indicated in Figure 1A, when apparatus 10 is assembled, conical member 22 is inserted, at least partially, into container 12 and the closed bottom end 30 of conical member 22 is held above collection area 16 of container 12. A starting sample solution of seed 18 and liquid carrier / solute 20 is poured into internal space 24 of conical member 22. Impeller 36 is lowered into conical member 22 and is operatively connected to rotary power source 34 by the impeller interface 40. The impeller 36 is configured to influence rotation of the starting solution to and / or toward the internal pore-bearing surface 26 of conical member 22.

[0152] As shown in Figure 1A, impeller 36 can include an impeller body 38 and an interface 40, wherein the interface 40 is configured to operatively connect the impeller 36 to the rotary power source 34. The impeller 36 can comprise any variety of form factors such that it is effective to influence rotation of liquid and / or seed suspended therein. According to some embodiments, the impeller 36 can be and / or comprise the Bottle Brush Drill Accessory produced by RotoScrub 36A (ASIN: B075L5H4XK) (see www.rotoproducts.com / products / bottle-cleaning-brush (date accessed: October 18, 2024)) (see also www.amazon.com / dp / B075L5H4XK?ref=emc_s_m_5_i_atc (date accessed: October 18, 2024)). Example images of the Bottle Brush Drill Accessory produced by RotoScrub 36A are shown in Figures 7A-C. According to some embodiments, the Bottle Brush Drill produced by RotoScrub 36A can comprise a universal steel shaft that works with any drill; a long, durable, extended reach neck with a reinforced steel spine that is configured to be able to reach deep into containers; and / or flexible and / or resilient non-scratch, nylon bristles configured to be able to fit into openings as small as 1 inch in diameter. According to some embodiments, the Bottle Brush Drill produced by RotoScrub 36A can have the following dimensions: about 12 inches or aboutAgent Ref. No. P14771WOOO12.75 inches in length inclusive of the brush head, about 9 inches or about 9.75 inches in length exclusive of the brush head, about 2.3 to 3 inches in width, and about 3 to 3.75 inches in height (i.e., the height / length of the brush head). The Bottle Brush Drill produced by RotoScrub 36A can comprise steel, stainless steel, and / or plastic. The Bottle Brush Drill produced by RotoScrub 36A can include a 0.25 inch hex shank steel shaft with rust-resistant plating and can be used with any drill, impact driver, or rotary power source. According to some embodiments, the Bottle Brush Drill produced by RotoScrub 36A can weigh about 0.01 ounces.

[0153] According to some embodiments, the impeller 36 can be and / or comprise the Paint Mixer Drill Attachment produced by Edward Tools 36B (ASIN: B08TG6NGSN) (see www. amazon. com / Edward-Tool s-Paint-Mixer-Attachment / dp / B 08 T G6N GSN / ref=asc_df_B 08 T G6N GSN / ? tag=hy prod- 20&linkCode=df0&hvadid=693274643898&hvpos=&hvnetw=g&hvrand=l 1455096557745805 773&hvpone=&hvptwo=&hvqmt=&hvdev=c&hvdvcmdl=&hvlocint=&hvlocphy=9016722&hv targid=pla-1918314638158&mcid=d778edccl225374e8eal4f7dc597db0e&th=l (date accessed: October 18, 2024)). The Paint Mixer Drill Attachment produced by Edward Tools 36B can fit all 3 / 8 inch hand drills and larger sizes as well. The Paint Mixer Drill Attachment produced by Edward Tools 36B allows for quick cleaning and reuse. According to some embodiments, the Paint Mixer Drill Attachment produced by Edward Tools 36B can measure about 10.75 inches in length inclusive of the head, and the head can measure about 2.5 inches in width. The Paint Mixer Drill Attachment produced by Edward Tools 36B can be about 6.4 ounces in weight. An example image of the Paint Mixer Drill Attachment produced by Edward Tools 36B is shown in Figure 8.

[0154] According to some embodiments, the impeller 36 can be and / or comprise the 43420 Paint Mixer produced by Hyde 36C (ASIN: B000W68Z04) (see www.amazon.com / HYDE- TOOLS-43420-Gallon- Paint / dp / B000W68Z04 / ref=sr_l_2?crid=2KKSXWEEOXJJL&dib=eyJ2IjoiMSJ9.RA2hJjGWA hZMH3ZPvvqK94Am8YyBMYJYb3-NE9MU04SprhgGWTlKeFv0NHQwHxzB2CNVqj- sXJWHZpzdN5r9VK4FjPYrbWLey_tgs7DuOVce9fNqaHaOLFFmwkzl- u99NzhJo7wLNNSRln3H70KPaCqlN4IBlrjcnB_Efq4nmGppEso7Yro-tm5i7ogI- s9my9UzqofSXafYvYukQDxaa_opCghSXlYuu7ur8z0alVNKRRskFpW6JdnfH2CawoqvVlEP ynVCg9d5_qtu-UgyfSexji8VSRH2O4O7pMdf6kw.m0Q4dqHcmfIoBhD70ycKW5LlR3s2- HZJe9YkFpblulA&dib_tag=se&keywords=Hyde+Tools+43420+l+Gallon%2FQuart+Paint+Mi xer%2C+ 11 %E2%80%9D+x+ 1 %2F4&qid= 1726662803&sprefix=hyde+tools+43420+ 1 +gallon %2Fquart+paint+mixer%2C+l l+x+l%2F4%2Caps%2C256&sr=8-2#customerReviews (dateAgent Ref. No. P14771WOOO accessed: October 18, 2024)). The weight of the 43420 Paint Mixer produced by Hyde 36C can be 0.21 pounds. An example image of the 43420 Paint Mixer produced by Hyde 36C is shown in Figure 9. According to some embodiments, the 43420 Paint Mixer produced by Hyde 36C can have the following dimensions: about 11 inches in length, about 2.5 inches in width, and about 2.5 inches in height. The 43420 Paint Mixer produced by Hyde 36C can have a weight of about 3.2 ounces.

[0155] According to some embodiments, the impeller 36 can be and / or comprise the Drill Brush Power Scrubber by Useful Products - Drillbrush Soft White Automotive Cleaning kit with Extended Reach Attachment - Drill Bit Extension Carpet Cleaner Solution - Car Interior Brush Set 36D (ASIN: B08FNX6NVF) (Global Trade Identification Number: 00810003602848) (see www.amazon.com / Drill-Brush-Scrubber-Useful- Products / dp / B08FNX6NVF?ref_=ast_sto_dp&th=l (date accessed: October 18, 2024)). An example image of the Drill Brush Power Scrubber by Useful Products 36D is shown in Figure 10. According to some embodiments, the Drill Brush Power Scrubber by Useful Products 36D can include a 5-inch-long corner brush and original drill bit extender. According to some embodiments, the Drill Brush Power Scrubber manufactured by Useful Products 36D can be soft and extendable. The Drill Brush Power Scrubber manufactured by Useful Products 36D can comprise nylon bristles and a plastic handle. The Drill Brush Power Scrubber manufactured by Useful Products 36D can have a weight of about 15 ounces.

[0156] As is shown in Figure 1A, the impeller 36, when operational, can be positioned generally at least partially within the internal space 24 of the conical member 22. According to some embodiments, when operational, the impeller body 38 can be positioned above the closed opposite bottom end 30 of the conical member 22 (e.g., about 1 to 2 inches above the closed opposite bottom end 30). According to some embodiments, when operational, the impeller body 38 can be positioned about 1 to 2 inches from the bottom of the container 12. According to some embodiments, the impeller body 38 can be generally cylindrical.

[0157] The impeller body 38 can extend from a lower end 37 to an opposite end 39 along the rotational axis 45 of the impeller 36. The impeller body 38 can further include a structure that extends generally toward or to the internal surface 26 of the conical member 22. For example, according to various embodiments, the impeller body 38 can comprise bristles and / or a brush with bristles, such as a tire brush, that extends generally toward or to the internal surface 26 of the conical member 22. The bristles can extend generally laterally from the rotational axis 45 of the impeller 36 toward the internal surface 26 of the conical member 22 when in operative position, wherein at least some of the bristles extend to the internal surface 26. According toAgent Ref. No. P14771WOOO some embodiments that include a tire brush, the brush can include a drill adapter for removably attaching the impeller 36 to the rotary power source 34. According to various embodiments wherein the impeller body 38 comprises bristles and / or a brush, the bristles and / or brush can be generally cylindrical. According to some embodiments, the bristles and / or brush can be and / or comprise nylon. According to some embodiments, the bristles can be non-scratch, flexible bristles and can fit into openings less than 1 inch in diameter. According to some embodiments, the total diameter of the bristles can be about 2.3 to 3 inches. According to some embodiments, the perimeter of impeller body 38, including any bristles and / or brush thereof, can have as much contact with the conical member 22 as possible. For example, according to some embodiments, when in use, the impeller body 38 can have a cross-sectional diameter, at least towards its lower end 37, that is greater than the cross sectional diameter of internal surface 26 of conical member 22 when impeller body 38 is in operative position. In at least one non-limiting example, the surface 26 has a cross-sectional diameter of about 2.3 inches at approximately 1 inch above the closed opposite bottom end 30, and the impeller body 38 has a cross-sectional diameter of at least 2.3 inches so that this portion of the impeller body 38 abuts, touches, and / or compresses, at least slightly, against internal surface 26. According to at least one non-limiting example, the internal surface 26 has a cross-sectional diameter of approximately 2 inches at approximately 1 inch above the closed opposite bottom end 30. According to at least one non-limiting example, the impeller body 38 has a cross-sectional diameter of approximately 2.5 inches.

[0158] According to some embodiments, the impeller body 38 can be and / or comprise a structure of any kind of shape or configuration including, but not limited to, a spherical shape (such as a ball), bead(s), a cylindrical shape, or other shape.

[0159] The impeller interface 40 can be a structure extending along the rotational axis 45 of the impeller 36, wherein the impeller interface 40 is configured to operatively attach and / or connect the impeller 36 to the rotary power source 34 such that the rotary power source 34 can rotate the impeller 36 and / or cause the impeller 36 to rotate. For example, the impeller interface 40 can be an axle, a rod, a pole, and the like. The interface 40 and / or the rotary power source 34 can include a chuck, lock, or any other suitable releasable or fixed connection mechanism to facilitate operative connection between the impeller interface 40 and the rotary power source 34. The impeller interface 40 and / or rotary power source 34 can include a corresponding element such as a notch, groove, indentation, and the like to mate, interlock, and / or operatively attach in some way the interface 40 to the rotary power source 34.

[0160] The apparatus 10 is configured such that a combination and / or suspension of whole dicot seeds 18 and a liquid 20 can occupy and / or be introduced / poured into the internal space 24 ofAgent Ref. No. P14771WOOO the conical member 22 as is shown in Figure 1A. It is noted that reference numeral 20 in Figure 1A shows the top or crest of liquid 20. The whole seeds 18 can be the seeds of any dicot plant. For example, the whole seeds 18 can be any seeds of the Fabaceae family. As another example, the whole seeds 18 can be soybean seeds. The liquid 20 can be any liquid suitable for suspending a plurality of seeds. According to some embodiments, the number of whole dicot seeds 18 can number anywhere from 1 to more than 1,000. According to some embodiments, the whole dicot seeds 18 are sterile seeds. According to some embodiments, the liquid 20 is water. Additionally or alternatively, the liquid 20 is sterile water. Prior to being added to the apparatus 10, liquid 20 can be stored in media bottle(s) 43 wherein said media bottle(s) 43 can be litersized (e.g., about 5 liter bottle(s)) and / or can include a cap or other type of closure. An example media bottle 43 is shown in Figure 11. Additionally or alternatively, the media bottle(s) 43 can be substantially made of glass and / or they can be reusable. The amount of liquid 20 used by the apparatus 10 to perform embryonic axis extraction and collection for the whole dicot seeds 18, which can include multiple successions, can be stored in such bottle(s). Such an amount of liquid could be approximately 6 to 15 liters. According to some embodiments, the media bottle(s) 43 can be and / or comprise one or more Reusable Glass Media Bottles with Cap manufactured by Fisher Scientific(Catalog No.: FB8005000) (see www.fishersci.com / shop / products / fisherbrand-reusable-glass-media-bottles-cap- 13 / FB8005000?gclid=Cj0KCQjw3ZayBhDRARIsAPWzx8q4YddoIKMKTGt_39h8dG8zK- iNnVwlrgJW8mZonCBep7cIQHWQWCEaAnVmEALw_wcB&ef_id=CjOKCQjw3ZayBhDRA RIsAPWzx8q4YddoIKMKTGt_39h8dG8zK- iNnVwlrgJW 8mZonCBep7cIQHWQWCEaAnVmEALw_wcB :G: s&ppc_id=PLA_goog_20861 45674_81843404874_FB8005000_386247001342_l 152236925343296614&ev_chn=shop&s_ kwcid=AL!4428!3 !386247001342! ! !g!858464079802!&gad_source=l (date accessed: October 18, 2024)). For example, the media bottle 43 shown in Figure 11 is the Reusable Glass Media Bottles with Cap manufactured by Fisher Scientific. According to some embodiments, the Reusable Glass Media Bottles with Cap manufactured by Fisher Scientific can be clear; drip- free; autoclavable (to 140 degrees Celsius); linerless; made of borosilicate glass; and can include a clear, drip-free, replaceable polypropylene pour ring; a polypropylene screw cap; a GL 45 screw thread; a permanent white enamel marking spot; a convenient wide opening / mouth; and enhanced graduations. According to some embodiments, the Reusable Glass Media Bottles with Cap manufactured by Fisher Scientific can have the following dimensions: a capacity of about 5000 milliliters, a diameter of about 7.1 inches, an inner neck diameter of about 1.18 inches, a height of about 12.9 inches, and / or graduations of about 1000 to 4000 milliliters.Agent Ref. No. P14771WOOO

[0161] The apparatus 10 is further configured such that the rotary power source 34 rotates the impeller and / or causes the impeller 36 to rotate such that the impeller 36 rotates the suspension of whole dicot seeds 18 and liquid 20 and / or causes the suspension of whole dicot seeds 18 and the liquid 20 to rotate. According to some embodiments, the rotation of the suspension influences the whole dicot seeds 18 toward and / or to the internal surface 26 of the conical member 22. The influencing of the whole dicot seeds 18 toward and / or to the internal surface 26 of the conical member 22 can include, but is not limited to, centrifugal force(s), mechanical force(s), and / or gravitational force(s).

[0162] The apparatus 10 is configured to comminute the whole dicot seeds 18 in order to promote separation of the embryonic axis of each dicot seed 18 from the rest of the seed so that the substantially extracted / separated embryonic axes of the seeds 18 can be collected. By rotating the suspension of the liquid 20 and the whole dicot seeds 18, the apparatus 10 imposes stresses on the seeds 18, wherein such stresses can include, but are not limited to, friction, shear or impact stress(es), degradation, and the like. The comminution can comprise one or more forces including friction, shear or impact stress(es), and degradation, but the comminution is not limited to such forces. For example, comminution of the seeds 18 can occur based on the seeds’ 18 contact with the internal surface 26 of the conical member 22, contact with the impeller 36 and / or components thereof, contact with other seeds 18, force(s) and / or stress(es) acting on the seeds 18 due to rotation of the suspension of the seeds 18 and the liquid 20, and / or any combination thereof. According to some embodiments, the internal surface 26 and the impeller 36 together are configured to comminute and / or degrade the whole dicot seeds 18 from original whole seeds to into their physiological parts but effectively retain most embryonic axes intact and / or substantially intact.

[0163] The comminution of the whole seeds 18 provides substantial separation / extraction of the embryonic axis of each whole seed 18 from the rest of the seed. The rotation of the suspension then forces the extracted embryonic axes to pass through the through-pores 32 of the conical member 22 wherein the passed embryonic axes 42 can be collected at and / or near the collection area 16 of the container 12 as shown in Figure 1. The through-pores 32 are sized so that they can generally pass embryonic axes 42 (shown diagrammatically in Figure 1A as diamond shapes) without passing other parts of the seeds and / or seed debris. However, according to some embodiments, sometimes debris inadvertently passes through the through-pores 32.

[0164] It can be appreciated by the foregoing that apparatus 10 is configured to influence the starting suspension of whole seeds 18 and liquid 20 to rotate the suspension in conical member 22 to, in turn, impart stress(es) on the seeds 18 in the suspension to comminute (break down intoAgent Ref. No. P14771WOOO smaller pieces), in particular, controlling the comminution by (a) selection of impeller body 38 characteristics (e.g., size, shape, materials, etc.), the liquid of the suspension, relation of the impeller 38 to surface 26 of conical member 22, and size and characteristics of the one or more through-pores 32; and (b) control of speed and duration of time of rotating the impeller 36 to promote sufficient comminution of seeds 18 to effectively extract substantially intact embryonic axes 42 from the seeds 18.

[0165] Figure IB shows a side elevation transparent perspective view of an example embodiment of aspects of a conical member 722 and impeller 736. The conical member 722 can comprise an internal surface 726, an open top base end 728, and a closed opposite bottom end 730. Such components can be the same as and / or similar to corresponding components described herein such as those of the conical member 22, the conical member 122, the conical member 1222, the conical member 1322, and / or components thereof. The impeller 736 can comprise an impeller body 738 and an impeller interface 740. The impeller body 738 can comprise a lower end 737 and an opposite end 739. Such components can be the same as and / or similar to corresponding components described herein such as those of the impeller 36, the impeller 136, the impeller 1236, the impeller 1336, and / or components thereof. Additionally, according to some embodiments, the configuration of components shown in Figure IB can be included in the apparatus 10 and / or the apparatus 110. For example, according to some embodiments, the conical member 22 and / or the conical member 122 can be and / or comprise the conical member 722. Similarly, according to some embodiments, the impeller 36 and / or the impeller 136 can be and / or comprise the impeller 736. As shown in Figure IB, and as noted above, according to some embodiments, the impeller 736 can have a cross-sectional diameter, at least toward its lower end 737, that is greater than the cross-sectional diameter of the internal surface 726 of the conical member 722 at least where the impeller body 738 is in contact with the internal surface 726 when the components are in operative position. As shown in Figure IB, the impeller body 738 can abut, touch, be in contact with, and / or compress against the internal surface 726 of the conical member 722.

[0166] Figure 1C shows a side elevation transparent perspective view of an example embodiment of aspects of a conical member 822 and impeller 836. The conical member 822 can comprise an internal surface 826, an open top base end 828, and a closed opposite bottom end 830. Such components can be the same as and / or similar to corresponding components described herein such as those of the conical member 22, the conical member 122, the conical member 1222, the conical member 1322, and / or components thereof. The impeller 836 can comprise an impeller body 838 and an impeller interface 840. The impeller body 838 can comprise a lowerAgent Ref. No. P14771WOOO end 837, an opposite end 839, and bristles 841. Such components can be the same as and / or similar to corresponding components described herein such as those of the impeller 36, the impeller 136, the impeller 1236, the impeller 1336, and / or components thereof. Additionally, according to some embodiments, the configuration of components shown in Figure 1C can be included in the apparatus 10 and / or the apparatus 110. For example, according to some embodiments, the conical member 22 and / or the conical member 122 can be and / or comprise the conical member 822. Similarly, according to some embodiments, the impeller 36 and / or the impeller 136 can be and / or comprise the impeller 836. The impeller 836 of Figure 1C includes an impeller body 838 that abuts, touches, is in contact with, and / or compresses against the internal surface 826 of the conical member 822 when in operative position. Additionally, as shown in Figure 1C, the impeller body 838 can comprise bristles 841. Such bristles 841 can abut, touch, be in contact with, and / or compress against the internal surface 826 of the conical member 822 when in operative position While the impeller body 838 of Figure 1C is shown to be in contact with the internal surface 826 of the conical member 822, according to some embodiments, the impeller body 838 need not be in contact with the internal surface 826 of the conical member 822.

[0167] Figure ID shows a side elevation transparent perspective view of an example embodiment of aspects of a conical member 922 and impeller 936. The conical member 922 can comprise an internal surface 926, an open top base end 928, and a closed opposite bottom end 930. Such components can be the same as and / or similar to corresponding components described herein such as those of the conical member 22, the conical member 122, the conical member 1222, the conical member 1322, and / or components thereof. The impeller 936 can comprise an impeller body 938 and an impeller interface 940. The impeller body 938 can comprise a lower end 937 and an opposite end 939. Such components can be the same as and / or similar to corresponding components described herein such as those of the impeller 36, the impeller 136, the impeller 1236, the impeller 1336, and / or components thereof. Additionally, according to some embodiments, the configuration of components shown in Figure ID can be included in the apparatus 10 and / or the apparatus 110. For example, according to some embodiments, the conical member 22 and / or the conical member 122 can be and / or comprise the conical member 922. Similarly, according to some embodiments, the impeller 36 and / or the impeller 136 can be and / or comprise the impeller 936. The impeller 936 of Figure ID includes an impeller body 938 that abuts, touches, is in contact with, and / or compresses against the internal surface 926 of the conical member 922 when in operative position. As shown in Figure ID, according to some embodiments, the impeller body 938 can include flexible and / or rigid flaps and / or flanges 945Agent Ref. No. P14771WOOO wherein such flaps and / or flanges 945 abut, touch, are in contact with, and / or compress against the internal surface 926 of the conical member 922 when in operative position. While the impeller body 938 of Figure ID is shown to be in contact with the internal surface 926 of the conical member 922, according to some embodiments, the impeller body 938 need not be in contact with the internal surface 926 of the conical member 922.

[0168] Figure IE shows a side elevation transparent perspective view of an example embodiment of aspects of a conical member 1022 and impeller 1036. The conical member 1022 can comprise an internal surface 1026, an open top base end 1028, and a closed opposite bottom end 1030. Such components can be the same as and / or similar to corresponding components described herein such as those of the conical member 22, the conical member 122, the conical member 1222, the conical member 1322, and / or components thereof. The impeller 1036 can comprise an impeller body 1038 and an impeller interface 1040. The impeller body 1038 can comprise a lower end 1037 and an opposite end 1039. Such components can be the same as and / or similar to corresponding components described herein such as those of the impeller 36, the impeller 136, the impeller 1236, the impeller 1336, and / or components thereof. Additionally, according to some embodiments, the configuration of components shown in Figure IE can be included in the apparatus 10 and / or the apparatus 110. For example, according to some embodiments, the conical member 22 and / or the conical member 122 can be and / or comprise the conical member 1022. Similarly, according to some embodiments, the impeller 36 and / or the impeller 136 can be and / or comprise the impeller 1036. As shown in Figure IE, according to some embodiments, the impeller body 1038 can be conical in shape. While the impeller 1038 is shown to have a truncated lower end 1037, the lower end 1037 could comprise a true conical apex according to some embodiments. While the impeller body 1038 of Figure IE is shown to not be in contact with the internal surface 1026 of the conical member 1022, according to some embodiments, the impeller body 1038 can be in contact with the internal surface 1026 of the conical member 1022.

[0169] Figure IF shows a side elevation transparent perspective view of an example embodiment of aspects of a conical member 1122 and impeller 1136. The conical member 1122 can comprise an internal surface 1126, an open top base end 1128, and a closed opposite bottom end 1130. Such components can be the same as and / or similar to corresponding components described herein such as those of the conical member 22, the conical member 122, the conical member 1222, the conical member 1322, and / or components thereof. The impeller 1136 can comprise an impeller body 1138 and an impeller interface 1140. The impeller body 1138 can comprise a lower end 1037 and an opposite end 1039. Such components can be the same asAgent Ref. No. P14771WOOO and / or similar to corresponding components described herein such as those of the impeller 36, the impeller 136, the impeller 1236, the impeller 1336, and / or components thereof. Additionally, according to some embodiments, the configuration of components shown in Figure IF can be included in the apparatus 10 and / or the apparatus 110. For example, according to some embodiments, the conical member 22 and / or the conical member 122 can be and / or comprise the conical member 1122. Similarly, according to some embodiments, the impeller 36 and / or the impeller 136 can be and / or comprise the impeller 1136. As shown in Figure IF, according to some embodiments, the impeller body 1138 can resemble a propeller or similar shape and can include one or more blades 1145. The blades 1145 can be rigid and / or flexible according to various embodiments. While the impeller body 1138 of Figure IF is shown to be in contact with the internal surface 1126 of the conical member 1122, according to some embodiments, the impeller body 1138 need not be in contact with the internal surface 1126 of the conical member 1122

[0170] According to various embodiments, the apparatus 10 and / or the apparatus 110, both described herein, can incorporate any of the configurations of any of Figures 1B-1F and / or can incorporate any combinations of such configurations.

[0171] Figures 12-17 show views of a specific example embodiment of an embryonic axis extraction and collection apparatus 110. The apparatus 110 can be similar to and / or the same as the apparatus 10. According to some embodiments, the apparatus 110 can comprise the apparatus 10 and / or any components or elements thereof. The apparatus 110 can function in the same manner and / or can have any of the same characteristics as the apparatus 10. For example, like components of the apparatus 110 can be similar to and / or the same as like components of the apparatus 10.

[0172] Just as the apparatus 10, the apparatus 110 is configured to automatically, or at least semi-automatically, comminute whole dicot seeds 118 in liquid suspension in an inverted conical member 122 in order to promote separation of the embryonic axis of each dicot seed 118 from the rest of the seed so that the embryonic axes of the seeds 118 can be substantially separated / extracted and then collected. By rotating the suspension of the liquid 120 and the whole dicot seeds 118, the apparatus 110 imposes stresses on the seeds 118, wherein such stresses include, but are not limited to, friction, shear or impact stress(es), degradation, and the like. For example, comminution of the seeds 118 can occur based on the seeds’ 118 contact with the internal surface 126 of the conical member 122, contact with the impeller 136 and / or components thereof, contact with other seeds 118, force(s) and / or stress(es) acting on the seedsAgent Ref. No. P14771WOOO118 due to rotation of the suspension of the seeds 118 and the liquid 120, and / or any combination thereof.

[0173] The comminution of the whole seeds 118 provides separation / extraction of the embryonic axis of each whole seed 118 from the rest of the seed. The rotation of the suspension then forces the extracted embryonic axes to pass through the through-pore(s) 132 of the conical member 122 wherein the passed embryonic axes can be collected in the collection area 116, and / or any other suitable area, of the container 112. The through-pore(s) 132 are sized so that they can generally pass embryonic axes without passing other parts of the seeds and / or seed debris. However, according to some embodiments, sometimes debris inadvertently passes through the through-pores 132.

[0174] As shown in Figures 12-17, the apparatus 110 includes a container 112, a conical member 122, a rotary power source 134, and an impeller 136. The container 112 can include an internal volume 114 and a collection area 116. The conical member 122 can include an internal space 124, an internal surface 126, an open top base end 128, a closed opposite bottom end 130, and one or more through-pores 132. According to some embodiments, as noted above with respect to the closed opposite bottom end 130 (as shown in Figure 16 with a stopper 144), the closed opposite bottom end 130 can be naturally open such that materials, such as solid or liquid material, could enter and exit the internal space 124 of the conical member 122 via the closed opposite bottom end. In such embodiments wherein the closed opposite bottom end 130 is naturally open, a stopper 144, as shown in Figure 16, can be inserted at and / or into the closed opposite bottom end 130 to effectively close the closed opposite bottom end 130 such that no materials, such as solid or liquid materials, can enter and / or exit the internal space 124 of the conical member 122 via the closed opposite bottom end 130. The stopper 144 can be inserted at and / or into the closed opposite bottom end 130 via forceps and / or any other suitable manner. The stopper 144 can be any sort of stopper including, but not limited to, a rubber stopper, a plastic stopper, and the like. Additionally or alternatively, any sort of closing device / mechanism could be used in addition to or alternatively to the stopper 144. According to embodiments that include the stopper 144, the conical member 122 can be truncated such that the stopper effectively closes the closed opposite bottom end 130 at the truncation plane. Figure 15 shows a perspective view of an example stopper 144 separated from conical member 122. Figure 16 shows the stopper 144 inserted at the closed opposite bottom end 130 of the conical member 122. Again, as noted herein with reference to the embodiment depicted in Figures 19A-G, according to some embodiments, the conical member can have an originally closed opposite bottom end such that a stopper is not necessary.Agent Ref. No. P14771WOOO

[0175] According to some embodiments, an outer surface of the conical member 122 can include one or more raised domes at least partially covering each through-pore 132. Each dome can be configured to protrude outwardly from the outer surface of the conical member 122 such that each dome partially covers at least a portion of each through-pore 132. According to some embodiments, each through-pore 132 can comprise a raised dome such that the number of through-pores 132 is equal to the number of raised domes. Each dome can have a top and a bottom. The top of each dome can be operatively attached and / or in contact with the outer surface of the conical member 122, and the bottom of each dome can be generally open and / or unattached to the conical member 122 such that any materials (such as liquid and / or embryonic axes) that pass through the through-pores 132 can exit the internal space 124 of the conical member 122 and collect in the container 112. The top of each dome can be generally circular, spherical, and / or ovate in shape, however, any suitable shape could be used. The bottom of each dome could be generally straight such that the bottom comprises a cut-off and / or truncation line of a truncated sphere and / or ovoid shape. However, any suitable shape could be used. When the conical member 122 is in the operative, inverted position such that the open top base end 128 faces upward (i.e., positioned as the “top” end) and the closed opposite bottom end 130 faces downward (i.e., positioned as the “bottom” end) each dome can protrude on the outer surface of the conical member 122 at or near the top of each through-pore 132 and extend downward such that a top portion of each through-pore 132 is at least partially covered. The bottom of each through-pore 132 can be substantially uncovered. Each dome can be substantially circular, spherical, and / or ovate in shape. For example, according to some embodiments, each dome can comprise one quarter, and / or similar to one quarter, of a complete sphere and / or ovoid.According to some embodiments, the height of each dome, meaning the distance each dome extends from the outer surface of the conical member 122, can be about 1 to 2 millimeters. However, according to various embodiments, the height of each dome can vary based on factors including, but not limited to, the size of the seed and / or genotype, the size of the through-pores 132, and the like.

[0176] The apparatus 110 can further include a clamp 146 configured to secure the conical member 122 to the container 112 during rotation of the suspension of whole dicot seeds 118 and liquid 120. Securing the conical member 122 to the container 112 during rotation of the suspension of whole dicot seeds 118 can serve to fix the position of the conical member 122 relative to the container 112. The clamp 146 can be any suitable type of clamp capable of securing the conical member 122 to the container 112. For example, the clamp 146 could be any sort of fastening device including, but not limited to, a microscope clamp, C-clamp, G-clamp,Agent Ref. No. P14771WOOO bar clamp, F-clamp, holdfast, pipe clamp, clip(s), and the like. According to some embodiments, the clamp 146 can have an opening of 2 11 / 16 inches, a width of 1 / i inches, and a depth of 2 inches. According to some embodiments, the clamp 146 can be and / or comprise the clamp from the magnifying lamp Multi-Lens LED Magnifier manufactured by ULINE 146A (Model No. El- 8788 and / or H-8789) (see www.uline.com / Product / Detail / H-8788 / Magnifiers / Multi-Lens-LED- Magnifier-175-and-225-Lenses?pricode=WB9566&gadtype=pla&id=H- 8788&gad_source=l&gclid=EAIaIQobChMIg5v3k5JhwMVPzEIBR2hRgz_EAQYASABEgL nsPD BwE (date accessed: October 18, 2024)). An example image of the clamp from the magnifying lamp Multi-Lens LED Magnifier produced by ULINE 146A is shown in Figure 18.

[0177] According to some embodiments, the clamp 146 is configured to secure the conical member 122 to the container 112 at a particular angle. For example, the clamp 146 can be configured to secure the conical member 122 to the container 112 wherein the conical member 122 is angled away from the rotary power source 134 such that the longitudinal axis 123 of the conical member 122, which extends from the open top base end 128 to the closed opposite bottom end 130, is not at a 90 degree angle relative to the platform and / or table upon which it rests. According to some embodiments, the clamp 146 is configured to releasably secure / attach the conical member 122 to the container 112 such that the longitudinal axis 123 of the conical member 122 is oblique to and / or offset from the rotational axis 145 of the impeller 136 and / or the longitudinal axis 113 of the container 112 when the apparatus 110 is operational. The angle at which the conical member 122 is secured to the container 112 during operation can be seen in Figure 17. As shown in Figure 17, the longitudinal axis 123 of the conical member 122 is oblique to and / or offset from the longitudinal axis 113 of the container 112 and to the rotational axis 145 of the impeller when the impeller 136 is in the operative position. Figure 17 shows the longitudinal axis 113 of the container 112 and the rotational axis 145 of the impeller 136 to be the same axis as, according to some embodiments, the longitudinal axis 113 of the container 112 and the rotational axis 145 of the impeller 136 are the same when the impeller 136 is in operative position. The longitudinal axis 123 of the conical member 122 and the longitudinal axis 113 of the container 112 / rotational axis 145 of the impeller 136 are shown to differ in Figure 17 by an angle a. According to some embodiments, the angle a can range from zero degrees to 90 degrees. As noted, according to some embodiments, the rotational axis 145 of the impeller 136 is parallel to and / or aligned with the longitudinal axis 113 of the container 112. Thus, in such embodiments when the longitudinal axis 123 of the conical member 122 is oblique to and / or offset from the longitudinal axis 113 of the container 112, the longitudinal axis 123 of the conical member 122 is also oblique to and / or offset from the rotational axis 145 of theAgent Ref. No. P14771WOOO impeller 136. According to some embodiments, the longitudinal axis 123 of the conical member 122, the longitudinal axis 113 of the container 112, and the rotational axis 145 of the impeller 136 could all be aligned. According to various embodiments, any combination of the longitudinal axis 123 of the conical member 122, the longitudinal axis 113 of the container 112, and / or the rotational axis 145 of the impeller 136 could be offset and / or aligned with one another. For example, the longitudinal axis 123 of the conical member 122 and the longitudinal axis 113 of the container 112 could be aligned and the rotational axis 145 of the impeller 136 could be oblique to and / or offset from the longitudinal axes 113, 123.

[0178] As shown in Figure 12, the rotary power source 134 can be operatively connected to a stand 148. The stand 148 can provide stability and ease of use when operating the apparatus 110. The stand 148 is configured to secure the rotary power source 134 and the container 112 in place, at least relative to each other, during operation. According to some embodiments, the stand 148 can include a base 150 for support, a securing member 152 configured to secure the container 112 in place and / or secure an elongated member 154 to the base 150, and an upper member 156 configured for releasably securing the rotary power source 134 and impeller 136 to the stand 148. It is noted that according to some embodiments, when the container is a 10,000 milliliter container and when the conical member is the conical member 1222 shown in Figure 19B, the securing member 152, including the first clamping portion 151, the second clamping portion 153, and the elongated portion 155, may not be included.

[0179] The base 150 of the stand can be any sort of generally flat platform upon which the container 112 can rest and which can support the stand 148 and other components. The base 150 is configured to properly position the container 112 relative to the rotary power source 134 such that the rotary power source 134 can interact with the impeller 136 to rotate and / or cause rotation of the suspension of whole dicot seeds 118 and the liquid 120. The base 150 can be placed on any flat surface such as the ground, the floor, a table, a chair, any sort of platform, and the like. For example, it can be beneficial, and is contemplated herein, to place the base 150 on the benchtop of a laminar flow hood, which ensures that the apparatus 110, and all portion(s) thereof and method(s) that can be used therewith, remain sterile. According to some embodiments, the base 150 can be placed on a non-flat surface.

[0180] The securing member 152 is configured to hold the container 112 in place relative to the stand 148. The securing member 152 can be operatively attached to both the container 112 and any portion of the stand 148 such as the elongated member 154 or the base 150. The securing member 152 can be any sort of fastening device capable of holding in place the container 112 relative to the stand 148. For example, the securing member 152 can be and / or comprise a utilityAgent Ref. No. P14771WOOO clamp and / or any other suitable clamping and / or securing device. According to some embodiments, the securing member 152 can comprise a first clamping portion 151, a second clamping portion 153, and an elongated portion 155. The first clamping portion 151 is configured to releasably attach to the container 112. The first clamping portion 151 can be and / or comprise any sort of clamp, clamping device, and / or securing device described herein as well as any sort of off-the-shelf clamp, clamping device, and / or securing device. The second clamping portion 153 is configured to releasably and / or slidingly attach to the elongated member 154 of the stand 148. The second clamping portion 153 can be and / or comprise any sort of clamp, clamping device, and / or securing device described herein as well as any sort of off-the- shelf clamp, clamping device, and / or securing device. The elongated portion 155 of the securing member 152 is configured to operatively connect the first clamping portion 151 and the second clamping portion 153. The elongated portion 155 can be any sort of rod, pole, and the like capable of operatively attaching to both the first and second clamping portions 151, 153 of the securing member 152. According to some embodiments, the height of the securing member 152 can be adjusted such that the securing member 152 can move up and / or down along the elongated member 154. One manner in which such adjustment of the height of the securing member 152 could be accomplished is by loosening the second clamping portion 153, sliding the securing member 152 up and / or down the elongated member 154, and then tightening the second clamping portion 153 to prevent inadvertent sliding of the securing member 152.

[0181] The elongated member 154 of the stand 148 can be and / or comprise any sort of elongated pole, rod, and / or any other long element. The elongated member can be operatively attached to the base 150, the securing member 152, the upper member 156 and / or the rotary power source 134. According to some embodiments, the elongated member 154 can be operatively attached to the base 150 at a lower end of the elongated member 154, and the elongated member 154 can extend upward from the base 150. The rotary power source 134 can be operatively attached to a top end of the elongated member 148 such that the rotary power source 134 can be positioned generally above the container 112. Thus, the impeller 136 can extend downward from the rotary power source 134 into the internal volume 114 of the container 112.

[0182] The upper member 156 can be configured to secure the rotary power source 134 to the stand 148. The upper member 156 can be operatively attached to both the rotary power source 134 and at least a portion of the stand 148 such as the elongated member 154 as is shown in Figure 12. The upper member 156 can be the same as and / or similar to the securing member 152. The upper member 156 can be any sort of fastening device capable of holding in place theAgent Ref. No. P14771WOOO rotary power source 134 relative to the stand 148. For example, the upper member 156 can be and / or comprise a utility clamp and / or any other suitable clamping and / or securing device. According to some embodiments, the upper member 156 can comprise a first clamping portion (not shown), a second clamping portion 157, and an elongated portion 159. The first clamping portion of the upper member 156 can be configured to releasably attach to the rotary power source 134. The first clamping portion of the upper member 156 can be and / or comprise any sort of clamp, clamping device, and / or securing device described herein as well as any sort of off- the-shelf clamp, clamping device, and / or securing device. According to some embodiments, the first clamping portion of the upper member 156 can be the same as and / or similar to the first clamping portion 151. The second clamping portion 157 can be configured to releasably and / or slidingly attach to the elongated member 154 of the stand 148. The second clamping portion 153 can be and / or comprise any sort of clamp, clamping device, and / or securing device described herein as well as any sort of off-the-shelf clamp, clamping device, and / or securing device. According to some embodiments, the second clamping portion 157 can be the same as and / or similar to the second clamping portion 153. The elongated portion 159 of the upper member 156 can be configured to operatively connect the first clamping portion of the upper member 156 and the second clamping portion 157. The elongated portion 159 can be any sort of rod, pole, and the like capable of operatively attaching to both the first (unlabeled) and second 157 clamping portions of the upper member 156. According to some embodiments, the height of the upper member 156 can be adjusted such that the upper member 156 can move up and / or down along the elongated member 154. One manner in which such adjustment of the height of the upper member 156 could be accomplished is by loosening the second clamping portion 157, sliding the upper member 156 up and / or down the elongated member 154, and then tightening the second clamping portion 157 to prevent inadvertent sliding of the upper member 156. When the rotary power source 134 is attached to the upper member 156, adjustment of the height of the upper member 156 serves to adjust the height of the rotary power source 134.

[0183] As disclosed with reference to the apparatus 10, the apparatus 110 can include an impeller 136 operatively attached to the rotary power source 134. The impeller can comprise an impeller body 138 and an interface 140 to connect to the rotary power source 134. As shown in Figures 12 and 14, the impeller body 138 can comprise bristles 141. The bristles 141 can extend generally laterally from the rotational axis 145 of the impeller 136 toward and / or to the internal surface 126 of the conical member 122. According to some embodiments, the bristles 141 and / or brush can be and / or comprise nylon. According to some embodiments, the impeller body 138, including any bristles and / or brush thereof, can have as much contact with the conical memberAgent Ref. No. P14771WOOO122 as possible. For example, according to some embodiments, when in use, the impeller body 138 could be positioned less than 1 inch from the closed opposite bottom end 130 of the conical member 122.

[0184] As shown in Figures 12 and 13, the rotary power source 134 and / or impeller 136 can include a securing mechanism 135 to releasably attach the impeller 136 to the rotary power source 134. The releasable securing mechanism 135 can be and / or comprise a chuck, lock, or any other suitable connection mechanism to facilitate operative connection between the impeller interface 140 and the rotary power source 134. The impeller interface 140 and / or rotary power source 134 can include a corresponding element such as a notch, groove, indentation, and the like to mate, interlock, and / or operatively attach in some way the interface 140 to the rotary power source 134.

[0185] According to some embodiments, the apparatus 110 can be used in combination with a sieve and a second container to further purify and / or collect embryonic axes 118 that have passed through the through-pores 132 and have been retrieved from collection area 116, or any other suitable area, of container 112.

[0186] Figures 19A-G show a partial assembly of an example embodiment of an embryonic axis extraction and collection apparatus 1210, and / or components thereof, according to at least some aspects of the present disclosure. With the exception of the conical member 1222, the example embodiment of Figures 19A-G can include all of the same and / or similar components as any extraction and / collection apparatus described herein including the apparatus 10 and / or the apparatus 110. While Figures 19A-G do not show a fully assembled apparatus, as shown in Figures 19A-G, the apparatus 1210 can include a container 1212 comprising a central longitudinal axis 1213, a conical member 1222 comprising a central longitudinal axis 1223, a rotary power source 1234, and an impeller 1236 comprising a rotational axis 1245. The longitudinal axis 1213 of the container 1212 and the rotational axis 1245 of the impeller 1236 are shown to be the same in Figure 19A since the two axes 1213, 1245 can be aligned according to some embodiments. As shown in Figure 19A, according to some embodiments, the longitudinal axis 1223 of the conical member 1222 can be oblique to and / or offset from the axes 1213, 1245 when the apparatus 1210 is in operative position. As noted herein, according to various embodiments, any combination of the axes 1212, 1223, 1245 can be aligned and / or offset with each other. As shown in Figure 19A, the container 1212 can include an internal volume 1214 and a collection area 1216. As shown in Figure 19A, the apparatus 1210 can include a securing mechanism 1235 to releasably attach the impeller 1236 to the rotary power source 1234. As shown in Figure 19A, the apparatus 1210 can include a clamp 1246 forAgent Ref. No. P14771WOOO releasably securing the conical member 1222 to the container 1212. As shown in Figure 19A, the apparatus can include a stand 1248 that comprises a base 1250, a securing member 1252, an elongated member 1254, and an upper member 1256. The upper member 1256 can include a first clamping portion (not shown), a second clamping portion 1257, and an elongated portion 1259. As noted, with the exception of the conical member 1222, each component of the apparatus 1210 can be the same and / or similar as any like component described herein. For example, the container 1212 can be the same as and / or similar to the container 12 and / or the container 112. Additionally, the apparatus 1210 can be used in any manner described herein such as via the method 310, the method 410, the method 510, and / or any other suitable method.

[0187] As shown in Figures 19A-G, the conical member 1222 can include an internal space 1224, an internal surface 1226, an outer surface 1227, an open top base end 1228, a closed opposite bottom end 1230, and at least one through-pore 1232. It is appreciated that for some embodiments that include the conical member 1222 shown in Figures 19A-G, less damage occurs to the meristem of the whole dicot seeds during operation and the extraction rate of the embryonic axes is 70 to 90% higher as compared to some other embodiments that do not include the conical member 1222. According to some embodiments, the conical member 1222 can comprise nylon 12.

[0188] The conical member 1222 can be customized via 3D-printing to match any specifications based on need or desire. As shown in Figures 19B-D, the conical member 1222 can be configured such that the through-pores 1232 cover substantially the entire surface 1226 of the conical member 1222. According to other embodiments, the through-pores of the conical member may not cover the entire surface of the conical member, but rather only a fraction of the surface such as one-third of the surface. By having through-pores 1232 that cover substantially the entire surface 1226 of the conical member 1222, the apparatus 1210 can provide greater through-put and be more effective and efficient than other embodiments. According to some embodiments, any suitable material such as metal material, plastic material, polymer material, resin material, wax material, and / or any other type of material capable of maintaining the proper structural integrity of the conical member 1222 could be used via 3D-printing to construct the conical member 1222.

[0189] The customizable nature of the conical member 1222, due at least in part to the conical member 1222 being 3D-printed, allows for several other benefits. The size and / or shape of the through-pores 1232 can be customized such that the size and shape of the through-pores 1232 can be optimized for different seeds and / or genotypes with different sized embryonic axes. Additionally, the conical member 1222 can hold a greater number of whole dicot seeds asAgent Ref. No. P14771WOOO compared to conical members of other embodiments. Further, the conical member 1222 is fully autoclavable. Even further, the conical member 1222 is easier to clean than conical members of other embodiments.

[0190] Additionally, the customizable nature of the conical member 1222 allows for the dimensions of the conical member 1222 to be customized based on need or desire. Figures 19B- G show various dimensions of aspects of the example conical member 1222. As shown in Figure 19B, the closed opposite bottom end 1230 of the conical member 1222 can be about 2.25 inches in diameter (as shown by length 1270 in Figure 19B), however, the closed opposite bottom end 1230 could have any suitable diameter depending on need and / or desire. As shown in Figure 19B, the closed opposite bottom end 1230 of the conical member 1222 can be originally closed such that a stopper and / or any other type of closure device and / or mechanism is not necessary to close the opposite bottom end 1230. As noted herein, according to some embodiments, the opposite bottom end 1230 of the conical member 1222 can be originally open such that a stopper and / or any other type of closing device and / or mechanism is necessary to close the opposite bottom end 1230.

[0191] As shown in Figure 8C, the open top base end 1228 of the conical member 1222 can have a diameter of about 8 inches (as shown by length 1272 in Figure 19C), however, the open top base end 1228 could have any suitable diameter depending on need and / or desire.

[0192] As shown in Figure 8D, the diagonal length of the conical member 1222 can be about 10.375 inches (as shown by length 1274 in Figure 19D), however, the conical member 1222 could have any suitable length depending on need and / or desire.

[0193] As shown in Figures 19B-D, the open top base end 1228 of the conical member 1222 can include a rim 1260 extending generally circularly around the perimeter of the open top base end 1228 of the conical member 1222. As shown in Figures 19B-D, the rim 1260 can extend radially outwardly from the outer surface 1227 and / or the open top base end 1228 of the conical member 1222. According to some embodiments, the conical member 1222, including the surface between the open top base end 1228 and the closed opposite bottom end 1230 can be about 5 millimeters thick. However, according to various embodiments, the thickness of the conical member 1222 can vary depending on need and / or desire. According to some embodiments, the rim 1260 can be about 8 millimeters thick. However, according to various embodiments, the thickness of the rim 1260 can vary depending on need and / or desire.

[0194] As shown in Figures 19B-G, each through-pore 1232 can include a raised dome 1233. Each dome 1233 can be configured to protrude outwardly from the outer surface 1227 of the conical member 1222 such that each dome 1233 partially covers at least a portion of eachAgent Ref. No. P14771WOOO through-pore 1232. Each dome 1233 can have a top 1237 and a bottom 1239. The top 1237 of each dome can be operatively attached and / or in contact with the outer surface 1227 of the conical member 1222 and the bottom 1239 of each dome 1222 can be generally open and / or unattached to the conical member 1222 such that any materials (such as liquid and / or embryonic axes) that pass through the through-pores 1232 can exit the internal space 1224 of the conical member 1222 and collect in the container 1212. The top 1237 of each dome 1233 can be generally circular, spherical, and / or ovate in shape, however, any suitable shape could be used. The bottom 1239 of each dome 1233 could be generally straight such that the bottom 1239 comprises a cut-off and / or truncation line of a truncated sphere and / or ovoid shape. However, any suitable shape could be used. As shown in Figures 19C and 19D, when the conical member 1222 is in the operative, inverted position such that the open top base end 1228 faces upward (i.e., positioned as the “top” end) and the closed opposite bottom end 1230 faces downward (i.e., positioned as the “bottom” end) each dome 1233 can protrude on the outer surface 1227 of the conical member 1222 at or near the top of each through-pore 1232 and extend downward such that a top portion of each through-pore 1232 is at least partially covered. The bottom of each through-pore 1232 can be substantially uncovered. As shown in Figures 19B-G, each dome 1233 can be substantially circular, spherical, and / or ovate in shape. For example, according to some embodiments, each dome 1233 can comprise one quarter, and / or similar to one quarter, of a complete sphere and / or ovoid.

[0195] Figures 19E-G show various dimensions of the through-pores 1232 and / or domes 1233. As shown in Figure 19E, each through-pore 1232 can be generally circular and can have a diameter of about 6 millimeters (as shown by length 1276 in Figure 19E). However, according to various embodiments, the through-pores 1232 can be any suitable shape and / or size based on need and / or desire.

[0196] As shown in Figure 19F, the length of each dome 1233, from the top 1237 to the bottom 1239, can be about 9 millimeters (as shown by length 1278 in Figure 19F). Thus, the top 1237 of each dome 1233 can be operatively attached to the outer surface 1227 of the conical member 1222 above the top of each through-pore 1232. According to various embodiments, the length of each dome 1233 can vary depending on the size of the through-pores 1232 and / or the positioning of each dome 1233 relative to each through-pore 1232.

[0197] As shown in Figure 19G, the height of each dome 1233, meaning the distance each dome 1233 extends from the outer surface 1227 of the conical member 1222, can be about 2 millimeters (as shown by the length 1280 in Figure 19G). However, according to variousAgent Ref. No. P14771WOOO embodiments, the height of each dome 1233 can vary based on factors including, but not limited to, the size of the seed and / or genotype, the size of the through-pores 1232, and the like.

[0198] Figure 20 shows a perspective view of a partial assembly of an example embodiment of an embryonic axis extraction and collection apparatus 1310, and / or components thereof, according to at least some aspects of the present disclosure. With the exception of the impeller 1336, the example embodiment of Figure 20 can include all of the same and / or similar components as any extraction and / collection apparatus described herein including the apparatus 10, the apparatus 110, and / or the apparatus 1210. The apparatus 1310 can incorporate any aspect(s) of any embryonic extraction apparatus described herein, such as the apparatus 10, the apparatus 110, and / or the apparatus 1210. While Figure 20 does not show a fully assembled apparatus, as shown in Figure 20, the apparatus 1310 can include a container 1312 comprising a spout 1315 and a longitudinal axis 1313, a conical member 1322 comprising a longitudinal axis 1323, a rotary power source 1334, and an impeller 1336 comprising a rotational axis 1345. In Figure 20, since, according to some embodiments, the longitudinal axis 1313 of the container 1312, the longitudinal axis 1323 of the conical member 1322, and the rotational axis 1345 of the impeller 1336 are aligned, each of the axes 1312, 1323, and 1345 are shown as a single axis.However, as noted herein, according to various embodiments, any combination of the axes 1312, 1323, 1345 can be aligned and / or offset with each other. The conical member 1322 can comprise an internal space 1324, an internal surface 1326, an open top base end 1328, and a plurality of through-pores 1332. As shown in Figure 20, the apparatus 1310 can include a stand 1348 that comprises at least a base 1350 and an elongated member 1354. The stand 1348 can be the same as and / or similar to any stand described herein including the stand 148 and / or the stand 1248. The stand 1348 can incorporate any aspect(s) of any stand described herein such as the stand 148 and / or the stand 1248.

[0199] As noted, with the exception of the impeller 1336, each component of the apparatus 1310 can be the same and / or similar as any like component described herein. For example, the container 1312 can be the same as and / or similar to the container 12, the container 112, and / or the container 1212. As another example, the conical member 1322 can be the same as and / or similar to any conical member described herein such as the conical member 22, the conical member 122, the conical member 722, the conical member 822, the conical member 922, the conical member 1022, the conical member 1122, and / or the conical member 1222. According to some embodiments, the conical member 1322 is and / or comprises the conical member 1222. Additionally, the apparatus 1310 can be used in any manner described herein such as via the method 310, the method 410, the method 510, and / or any other suitable method.Agent Ref. No. P14771WOOO

[0200] Figure 21 shows a perspective view of the impeller 1336 in isolation. According to various embodiments, any impeller described herein, including at least the impeller 36, the impeller 136, the impeller 736, the impeller 836, the impeller 936, the impeller 1036, the impeller 1136, and / or the impeller 1236, can be and / or comprise the impeller 1336 as shown in Figure 21. Further, the impeller 1336 can incorporate any aspect(s) of any impeller described herein, such as the impeller 36, the impeller 136, the impeller 736, the impeller 836, the impeller 936, the impeller 1036, the impeller 1136, and / or the impeller 1236.

[0201] As shown in Figure 21, according to some embodiments, the impeller 1336 can comprise an impeller body 1338 and an interface 1340 to connect to the rotary power source 1334. As shown in Figure 21, the impeller body 1338 can comprise a lower end 1337 and an opposite end 1339. The impeller body 1338 can further comprise one or more flanges 1342. According to some embodiments, the impeller 1336 can be 3D printed.

[0202] According to some embodiments, the impeller body 1338 can be the same as and / or similar to any impeller body described herein, can function in the same and / or similar manner as any impeller body described herein, and / or can incorporate any aspect(s) of any impeller body described herein. For example, the impeller body 1338 can extend from a lower end 1337 to an opposite end 1339 along the rotational axis 1345 of the impeller 1336. The impeller body 1338 can further include one or more flanges 1342 that extend generally toward or to the internal surface 1326 of the conical member 1322. As shown in Figure 21, according to some embodiments, the impeller body 1338 can comprise 6 flanges 1342. However, according to various embodiments, the impeller body 1338 can comprise any suitable number of flanges ranging from 1 to N where N is any number greater than 1. The one or more flanges 1342 can extend generally laterally from the rotational axis 1345 of the impeller 1336 toward or to the internal surface 1326 of the conical member 1322 when in operative position. The impeller 1336 can be designed to fit into the conical member 1322, which, according to some embodiments, is and / or comprises the conical member 1222.

[0203] As shown in Figure 21, according to some embodiments, the impeller body 1338, including the one or more flanges 1342, can be generally conical wherein the lower end 1337 represents the apex of a cone and the opposite end 1339 represents the base of the cone. In other words, the lateral extension of the one or more flanges 1342 at the lower end 1337 of the impeller body 1338 is less than the lateral extension of the one or more flanges 1342 at the opposite end 1339 of the impeller body 1338. Further, as shown in Figure 21, each flange 1342 of the one or more flanges 1342 can be rounded. According to various embodiments, the one orAgent Ref. No. P14771WOOO more flanges 1342 can be flexible or rigid. According to some embodiments, the one or more flanges 1342 can be resilient.

[0204] According to some embodiments, the perimeter of impeller body 1338, including the one or more flanges 1342 thereof, can have as much contact with the conical member 1322 as possible. According to some embodiments, when in use, the impeller body 1336 can have a cross-sectional diameter, at least towards its opposite end 1339, that is greater than the cross- sectional diameter of the internal surface 1326 of conical member 1322 when the impeller body 1338 is in operative position. In at least one non-limiting example, the internal surface 1326 has a cross-sectional diameter at approximately 1 inch above the bottom end of the conical member 1322 that is smaller than the cross-sectional diameter of the impeller body 1338, at least at the opposite end 1339, so that at least a portion of the impeller body 1338 abuts, touches, and / or compresses, at least slightly, against internal surface 1326.

[0205] According to some embodiments, the interface 1340 can be the same as and / or similar as any impeller interface described herein, can function in the same and / or similar manner as any impeller interface described herein, and / or can incorporate any aspect(s) of any impeller interface described herein. For example, the interface 1340 can be a structure extending along the rotational axis 1345 of the impeller 1336, wherein the interface 1340 is configured to operatively attach and / or connect the impeller 1336 to the rotary power source 1334, such that the rotary power source 1334 can rotate the impeller 1336 and / or cause the impeller 1336 to rotate. For example, the interface 1340 can be an axle, a rod, a pole, and the like. The interface 1340 and / or the rotary power source 1334 can include a chuck, lock, or any other suitable releasable or fixed connection mechanism to facilitate operative connection between the interface 1340 and the rotary power source 1334. The interface 1340 and / or rotary power source 1334 can include a corresponding element such as a notch, groove, indentation, and the like to mate, interlock, and / or operatively attach in some way the interface 1340 to the rotary power source 1334

[0206] Figure 22 shows a perspective view of the container 1312 wherein the container 1312 includes a cover 1360. Figure 22 also shows that the container 1312 can include a handle 1317. The cover 1360 can be releasably attached to the open, top portion of the container 1312 when the container 1312 is being autoclaved. As shown in Figure 22, according to some embodiments, the cover 1360 can be secured to the container 1312 via use of securing means 1364A. As shown in Figure 22, the securing means 1364A can be tape. However, the securing means 1364A can be any means capable of securing the cover 1360 to the container 1312 including, but not limited to, other adhesives, clamp(s), clasp(s), and the like. The cover 1360Agent Ref. No. P14771WOOO helps to eliminate the need to use aluminum foil or some other material to cover the open, top of the container 1312 when autoclaving the container 1312.

[0207] Figure 23 shows a top elevation view of the container 1312 wherein the container 1312 includes the cover 1360. As shown in Figure 23, the cover 1360 can comprise an aperture 1362. Figure 23 shows the interface 1340 of the impeller 1336 extending through the aperture 1362. The aperture 1362 of the cover 1360 allows the impeller 1336 to be, at least partially, within the container 1312 when the container 1312 is being autoclaved.

[0208] Figure 24 shows a perspective view of the container 1312 wherein the container 1312 includes a lip cover 1370 to cover the spout 1315. The lip cover 1370 can be releasably attached to the spout 1315. According to some embodiments, the lip cover 1370 can be secured to the spout 1315 via securing means 1364B. As shown in Figure 24, the securing means 1364B can be tape. However, the securing means 1364B can be any means capable of securing the cover 1360 to the container 1312 including, but not limited to, other adhesives, clamp(s), clasp(s), and the like. The lip cover 1370 can be used during embryonic extraction. The lip cover 1370 can be configured to cover the spout 1315 during embryonic extraction to prevent a user from contacting the spout 1315 during extraction, which helps to prevent contamination.

[0209] Figure 25 shows a perspective view of the impeller 1336 and the Paint Mixer Drill Attachment produced by Edward Tools 36B shown in Figure 8, both in isolation. The impeller 1336 is more efficient at breaking up seeds than the Paint Mixer Drill Attachment produced by Edward Tools 36B. For example, use of the impeller 1336 leads to lower extraction time of embryonic axes as compared to use of the Paint Mixer Drill Attachment produced by Edward Tools 36B. Further, use of the impeller 1336 leads to not having to mix / stir manually as frequently as compared to the Paint Mixer Drill Attachment produced by Edward Tools 36B.

[0210] Figure 26 shows a view of a sectioned dicot seed 210 with two sectioned opposite halves 211 A and 21 IB to show its constituent parts. Any dicot seeds described herein such as the dicot seeds 18, the dicot seeds 118, and / or any dicot seeds described with reference to any method disclosed herein can be and / or comprise the dicot seed 210 shown in Figure 26. The dicot seed 210 comprises a seed coat that is sectioned in halves 212A and 212B, two cotyledons 214, 215, and an embryonic axis 216. When the apparatus 10 and / or the apparatus 110 is in operation, the apparatuses 10, 110 comminute and / or degrade whole dicot seeds 210 to substantially separate and / or extract the embryonic axis 216 from each seed 210. While Figure 26 shows a view of a sectioned dicot seed 210 wherein the two cotyledons 214, 215 are broken apart, the two cotyledons of an intact dicot seed will surround the embryonic axis 216. Thus, the dicot seedsAgent Ref. No. P14771WOOO must be comminuted, degraded, and / or otherwise manipulated in order to substantially isolate the embryonic axes 216.

[0211] According to various embodiments, any comminution and / or degradation of whole dicot seeds described herein can include the relatively hard outer seed coat (hull) 212A, 212B being removed from the relatively soft tissue that the seed coat 212A, 212B encloses. Any comminution and / or degradation described herein can further include friction causing the two cotyledons 214, 215 of each dicot seed to unfold wherein the attachment between the embryonic axis 216 and the cotyledons 214, 215 eventually ruptures.

[0212] Additionally or alternatively, friction generated by the impeller body moving against the conical member can cause the seed coats of the whole dicot seeds to wear down, comminute, and / or degrade, thereby removing each seed coat from each whole dicot seed. Once the seed coat is removed, each dicot seed is able to split in half wherein each embryonic axis has a natural divide from the half seeds.

[0213] According to some embodiments, any comminution and / or degradation of whole dicot seeds described herein can be due to rotation of the suspension, contact between the whole dicot seeds and the impeller body (which can include a brush and / or bristles), contact between the whole dicot seeds and the conical member, contact between the whole dicot seeds themselves, friction generated by the impeller body moving against the conical member, and / or any combination thereof.

[0214] Figure 27 shows a flow chart illustrating a method 310 for automated, or at least semiautomated, extract! on / separati on and collection of embryonic axes of one or more whole dicot seeds. According to various embodiments, the method 310 can be performed via the apparatus 10, the apparatus 110, the apparatus 1210, the apparatus 1310, the system 610 described herein, and / or any component(s) thereof. Additionally or alternatively, the method 310 can incorporate any aspect(s) of any other method described herein. The method 310 comprises a first step 312 comprising rotating a suspension of a plurality of whole dicot seeds and a liquid relative to a surface having through-pores, wherein the surface is effective to: (i) comminute the whole dicot seeds into smaller pieces, and (ii) substantially pass embryonic axes. The plurality of whole dicot seeds and the liquid can be suspended in the internal space of the conical member that is positioned at least partially within the internal volume of the container as described by any embryonic axis extraction and collection apparatus included herein. In such embodiments, the surface can be the internal surface / portion of the conical member. Additionally or alternatively, the whole dicot seeds and the liquid could be suspended relative to any suitable surface having through-pores. The rotation of the suspension can be performed via a rotary power sourceAgent Ref. No. P14771WOOO operating in conjunction with an impeller. Additionally or alternatively, the rotation of the suspension can be performed by any suitable means capable of rotating a suspension of whole dicot seeds and a liquid such as magnetic steering.

[0215] The method 310 comprises a second step 314 comprising collecting the embryonic axes that have passed through the through-pores. As noted above regarding the various embryonic axis extraction and collection apparatuses described herein, such collection of the passed embryonic axes can occur via the collection area of the container. Additionally or alternatively, the embryonic axes that pass through the through-pores could be collected via any suitable collection device such as a bowl, bucket, pan, dish, and the like.

[0216] As noted above, the comminution of the whole dicot seeds can impose upon the whole dicot seeds one or more of friction, shear or impact stress(es), and / or degradation. The method 310 can further comprise influencing the whole dicot seeds toward and / or to the surface while rotating the suspension. Additionally, as noted above, such influencing of the whole dicot seeds toward and / or to the surface can be caused by one or more of centrifugal force(s), mechanical force(s), and / or gravitational force(s).

[0217] As noted above, the surface of the step 312 can comprise an internal surface / portion of a conical member such as any of the conical members described herein. As described herein, the conical member can be three-dimensional and can be inverted and positioned at least partially within an internal volume of a container when operational. The conical member can have an internal space along an axis between an open top base end and a closed opposite bottom end. The internal surface / portion of the conical member can be located between the open top base end and the closed opposite bottom end.

[0218] It should be noted that, according to some embodiments, any steps of the method 310 can be repeated, if desired, to continue to substantially separate / extract embryonic axes from any whole seeds that remain and to collect the substantially separated / extracted embryonic axes.

[0219] Figure 28 shows a flow chart illustrating a method 410 for extracting / separating and collecting embryonic axes from one or more whole dicot seeds. According to various embodiments, the method 410 can be performed via the apparatus 10, the apparatus 110, the apparatus 1210, the apparatus 1310, the system 610 described below, and / or any component s) thereof. Additionally or alternatively, the method 410 can incorporate any aspect(s) of any other method described herein. The method 410 comprises a first step 412 comprising securing a conical member to a container via a clamp. This step 412 can be performed via any of the components of any apparatus(es) and / or system(s) described herein. For example, the conical member, container, and clamp can be any such component described herein. As mentionedAgent Ref. No. P14771WOOO above, according to some embodiments, the conical member is configured to be secured to the container at a particular angle. For example, the conical member is configured to be secured to the container wherein the conical member is angled away from the rotary power source such that the longitudinal axis of the conical member is not at a 90 degree angle relative to the platform and / or table upon which it rests. According to some embodiments, the clamp is configured to releasably secure / attach the conical member to the container such that the longitudinal axis of the conical member is oblique to and / or offset from the rotational axis of the impeller and / or is oblique to and / or offset from the longitudinal axis of the container when the apparatus is operational.

[0220] The method 410 further comprises a step 414 comprising rotating, via an impeller, a suspension of a plurality of imbibed whole dicot seeds and a liquid relative to the conical member having through-pores, the conical member being effective to: (i) comminute the imbibed whole dicot seeds into smaller pieces, and (ii) substantially pass embryonic axes. This step can be performed in the same and / or similar manner as that described above with respect to step 312 of the method 310. Again, any components of any apparatus(es) and / or system(s) described herein can be used to perform this step 414. For example, the impeller could be any impeller described herein. The impeller could be operatively connected to a rotary power source that rotates the impeller and / or causes the impeller to rotate. As noted herein, the height of the impeller can be adjustable relative to the conical member and / or any portion(s) thereof. For example, the height of the impeller can be adjustable relative to the conical member.

[0221] As noted herein, the impeller can comprise an impeller body that comprises a structure extending toward or to the internal surface / portion of the conical member when the apparatus is operational. According to some embodiments, the impeller body comprises one or more bristles extending generally laterally from the rotational axis of the impeller towards, at, and / or near the internal portion / surface of the conical member when the apparatus is operational.

[0222] The method 410 further includes the step 416 comprising influencing the whole dicot seeds toward and / or to the conical member while rotating the suspension. Steps 414 and 416 are performed simultaneously according to some embodiments. The rotation of the suspension can cause the influencing of the whole seeds toward and / or to the conical member. Again, any components of any apparatus(es) and / or system(s) described herein can be used to perform this step 416.

[0223] The method 410 further includes the step 420 comprising pouring, and / or introducing, the passed embryonic axes into a second container. Once the embryonic axes are passed through the through-pores of the conical member (i.e., “passed embryonic axes”), the passed embryonicAgent Ref. No. P14771WOOO axes can be poured, and / or introduced, into a second container where they are collected. The second container can be any suitable type of container such as any beaker, pitcher, bowl, pan, dish, and the like. According to some embodiments, the second container can be a 2 liter pitcher, such as the 2000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12A that is described herein. According to some embodiments, method 410 can further include an optional step comprising sieving the passed embryonic axes when pouring the contents of the first container into the second container. This optional step can be performed via a transition sieve wherein the transition sieve can be any suitable type of sieve, strainer, filter, screen, mesh material, and the like capable of separating particulate matter having differing characteristics such as being of differing size. For example, this transition sieve can be the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430 (item model number: Z072-SSM). An example image of the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430 is shown in Figures 32A-C, wherein Figure 32A shows a front elevation view of the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430, Figure 32B shows a side perspective view thereof, and Figure 32C shows a bottom perspective view thereof. According to some embodiments, the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430 can have a capacity of about 34.6 ounces, can be about 4.72 inches in diameter, about 5.4 inches in depth, and the handle can be about 9.64 inches in length. According to some embodiments, the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430 can be made with high-grade stainless steel and can be lightweight, anti-rust, durable, heat-resistant, and safe for use with high-temperature materials such as liquid(s). According to some embodiments, the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430 can be sturdy and stable such that it will not tip over, can include a bottom that is covered with thickened steel wire, can include a dense, fine mesh, can include a deep basket design to hold contents securely, and / or can include a side hook and / or a hook at the end of the handle. The Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430 can be smooth both inside and outside such that it would not hurt a user’s hands and such that it is easy to clean.

[0224] According to some embodiments, this transition sieve can be adapted such that it includes holes / apertures that are each about 6 millimeters in diameter. This sieving can be performed such that embryonic axes that have already passed through the through-pores of the conical member are then poured over the sieve wherein the sieve is configured to substantially allow the embryonic axes to pass through to the second container while substantially preventingAgent Ref. No. P14771WOOO seed debris such as seed coats, cotyledons, and the like from passing through. This optional step of sieving the embryonic axes when pouring the contents of the first container into the second container can provide further isolation of the embryonic axes from any seed debris that may have inadvertently passed through the through-pores of the conical member.

[0225] It should be noted that, according to some embodiments, any steps of the method 410 can be repeated, if desired, to continue to substantially separate / extract embryonic axes from any whole seeds that remain and to collect the substantially separated / extracted embryonic axes.

[0226] Figures 29A-29E show a flow chart of a method 510 for extracting / separating and collecting embryonic axes from one or more whole dicot seeds. According to various embodiments, the method 510 can be performed via the apparatus 10, the apparatus 110, the apparatus 1210, the apparatus 1310, the system 610 described below, and / or any component s) thereof. Additionally or alternatively, the method 510 can incorporate any aspect(s) of any other method described herein.

[0227] The method 510 includes the step 512 comprising imbibing whole dicot seeds. According to at least some embodiments, the one or more whole dicot seeds are imbibed on solid media for 7 hours and then imbibed in water, which could be sterile water, for 14 hours. However, the whole dicot seeds can be imbibed via any suitable media for any suitable amount of time. According to some embodiments, imbibing the whole dicot seeds helps to soften the whole dicot seeds so that they are more easily comminuted. Additionally or alternatively, imbibing the whole dicot seeds leads to more effective use of the embryonic axes extracted from the whole dicot seeds. According to some embodiments, the whole dicot seeds can be imbibed for a period of time ranging from about 14 to about 23 hours. Imbibing the seeds can include, but is not limited to, soaking the seeds, causing moisture to absorb or assimilate into the seeds, and the like. According to some embodiments, the step 512 can include imbibing the seeds inside of a sterile container, such as a sterile flask, and placing the container and / or the seeds on a shaker mechanism. The shaker mechanism could be any sort of device and / or mechanism capable of moving, rotating, vibrating, and / or shaking a container such as a flask. According to some embodiments, the step 512 comprising imbibing the seeds includes imbibing the seeds inside of a sterile flask, wherein the sterile flask is placed on a shaker for 21 hours.

[0228] The method 510 further includes the step 514 comprising autoclaving an impeller, and / or components thereof. Any suitable method of autoclaving can be used such as high pressure, low pressure, high temperature, and / or low temperature. The impeller can be any impeller described herein. For example, the impeller can include an impeller body and an interface to operatively attach to a rotary power source. The impeller body can comprise bristles and / or a brush, such asAgent Ref. No. P14771WOOO a tire brush, according to some embodiments. Autoclaving the impeller, including the impeller body, can soften the bristles prior to performing the rest of the method. According to some embodiments, the bristles of the impeller body can undergo one or two autoclave cycles prior to use, which serves to the soften the bristles. This softening of the bristles leads to more effective use. According to some embodiments, the bristles and / or brush can be and / or comprise nylon.

[0229] The method 510 further includes the step 516 comprising sterilizing the impeller, and / or any components thereof. The impeller can be sterilized with application of chlorine gas according to some embodiments. However, any suitable method of sterilization could be used including, but not limited to, any sort of liquid and / or gas sterilization, bleach sterilization (such as 10% or 20% bleach solution), and the like. This step 516 comprising sterilizing the impeller can include sterilizing all parts of the impeller which, according to some embodiments, could include a brush and / or bristles. According to some embodiments, sterilization of the impeller, including a brush and / or bristles thereof, can occur wherein the impeller is attached to a rotary power source such that the rotary power source is spinning the impeller and / or causing the impeller to spin during sterilization. As noted, a 10% or 20% bleach solution could be used for sterilization, however, any suitable sterilizing material could be used depending on need and / or desire. For example, a 20% bleach solution could be used wherein the impeller is then washed with sterile water. Additionally or alternatively, according to some embodiments, the sterilization of the impeller stopper can occur the day before other steps of the method 510 are performed to extract and / or collect embryonic axes. According to various embodiments, the sterilization of the impeller could occur at any suitable time relative to performing the other steps of the method 510. According to some embodiments wherein a stopper is included, the step 516 can further comprise sterilizing a stopper wherein the stopper can be sterilized in the same manner as the impeller and / or in any manner described herein.

[0230] The method 510 further includes the step 518 comprising autoclaving at least three containers (at least a first container, second container, and third container), a conical member, and / or a liquid. Again, any suitable method of autoclaving can be used such as high pressure, low pressure, high temperature, and / or low temperature. Each of the containers could be and / or comprise any container described herein and / or any sort of container capable of containing plant seeds and a liquid including, but not limited to, a pitcher, beaker, bowl, pan, dish, and the like. For example, according to some embodiments, the first container can be and / or comprise the container 12, the container 112, the container 1212, and / or the container 1312. The conical member could be any conical member described herein such as the conical member 22, the conical member 122, the conical member 1222, and / or the conical member 1322, which allAgent Ref. No. P14771WOOO include an internal space, an internal surface, and at least one through-pore. The second container could be the same as and / or similar to the first container and / or any other container mentioned herein. The third container could be the same as and / or similar to the first container and / or any other container mentioned herein such as a petri dish. According to some embodiments, the liquid could be water and / or sterile water. Any suitable amount of liquid is contemplated herein. For example, the liquid could comprise about 6 to 15 liters. According to some embodiments, the step 518 can occur prior to the step 512.

[0231] The method 510 further includes the step 520 comprising placing a rotary power source in a laminar flow hood and sterilizing the rotary power source. The rotary power source could be any rotary power source described herein such as the rotary power source 34, the rotary power source 134, the rotary power source 1234, and / or the rotary power source 1334. Additionally, the rotary power source could be any suitable type of motor, engine, machine, and the like capable of rotating an impeller. The sterilization of the rotary power source can be performed by applying isopropyl alcohol (IP A) to the rotary power source. For example, such application could be performed by spraying and / or wiping down the rotary power source. The IPA could be 70% IPA and / or any other suitable IPA capable of sterilization. Once the rotary power source is sterilized, the height of the rotary power source can be adjusted such that the height is adjusted to its highest position. The rotary power source can be operatively connected to a stand and / or support structure such as the stand 148, the stand 1248, the stand 1348 described herein, and / or any other suitable stand. The stand and / or support structure can provide stability for the rotary power source, can aid in adjusting the height of the rotary power source, and can properly position the rotary power source relative to the first container. According to some embodiments, the step 520 can further comprise placing the stand and / or support structure in a laminar flow hood and / or sterilizing the stand and / or support structure in the same and / or similar manner as the rotary power source.

[0232] The method 510 further includes the step 521 comprising sterilizing a clamp. The clamp can be any clamp described herein and / or any other suitable type of clamp. The sterilization can be performed by applying isopropyl alcohol (IPA) to the clamp. Such application can be accomplished by spraying, wiping, and / or any other suitable manner of application. The IPA could be 70% IPA and / or any other suitable IPA capable of sterilization.

[0233] The method 510 further includes the step 522 comprising securing the conical member at least partially within the first container via the clamp, wherein the conical member is angled away from the rotary power source. The conical member can be secured such that it is positioned at least partially within an internal volume of the first container. The conical memberAgent Ref. No. P14771WOOO can be angled away from the rotary power source such that the longitudinal axis of the conical member is not at a 90 degree angle relative to the platform and / or table upon which it rests. According to some embodiments, the clamp is configured to releasably secure / attach the conical member to the first container such that the longitudinal axis of the conical member is oblique to and / or offset from the rotational axis of the impeller and / or is oblique to and / or offset from the longitudinal axis of the first container when the apparatus is operational

[0234] The method 510 further includes the step 524 comprising placing the impeller at least partially within the conical member. The impeller can be placed at least partially within the conical member with the impeller body end of the impeller facing downward.

[0235] The method 510 further includes the step 526 comprising operatively attaching the impeller to the rotary power source. The impeller should be attached to the rotary power source such that the rotary power source can rotate the impeller and / or cause rotation of the impeller. According to some embodiments, such attachment of the impeller to the rotary power source could be performed via any securing mechanism described herein such as the securing mechanism 135 and / or the securing mechanism 1235. According to some embodiments, the attachment of the impeller to the rotary power source comprises tightening a chuck, lock, and / or any other type of locking mechanism. According to some embodiments, a chuck could be tightened via a chuck key. According to some embodiments (such as those embodiments wherein the rotary power source is and / or comprises the 60L Overhead Stirrer manufactured by +UXI 34B and described herein), a chuck key may not be used to tighten a chuck and / or to connect the impeller to the rotary power source.

[0236] The method 510 further includes the step 528 comprising adjusting the height of the rotary power source as necessary. According to some embodiments, the height of the rotary power source should be adjusted such that the tire brush and / or bristles of the impeller body is / are positioned about 1 to 2 inches from a bottom of the conical member (such as the closed opposite bottom end of the conical member) and / or a bottom of the first container (such as the collection area of the first container). According to some embodiments, the height of the rotary power source can be adjusted such that the tire brush and / or bristles of the impeller body is / are positioned less than 1 inch from a bottom of the conical member (such as the closed opposite bottom end of the conical member) and / or a bottom of the first container (such as the collection area of the first container) such that the brush and / or bristles of the impeller body have as much contact with the conical member as possible. According to some embodiments, the height of the rotary power source can be adjusted such that the tire brush and / or bristles of the impeller body is / are positioned as low as possible without preventing rotation of the impeller, which can leadAgent Ref. No. P14771WOOO to high efficiency. According to some embodiments, the step 524 can comprise the steps 526 and 528.

[0237] The method 510 further includes the step 530 comprising powering ON the rotary power source to rotate the impeller and / or cause the impeller to rotate. The rotary power source can be powered ON and / or spun at a speed of about 75 to 500 RPM. According to some embodiments, the speed of the rotary power source is about 250 to 300 RPM. According to some embodiments, the speed of the rotary power source is about 170 to 200 RPM. According to some embodiments, the speed of the rotary power source is about 75 to 300 RPM. Additionally, when the rotary power source is powered ON, the step 530 can further comprise adjusting aspect(s) of any fasteners of the stand and / or support structure, such as the angle of any of the fasteners, such that the rotary power source does not move excessively when the rotary power source is powered ON. The fasteners of the stand and / or support structure can be and / or be similar to the securing member 152 and / or the upper member 156 of the stand 148.

[0238] The method 510 further includes the step 532 comprising adding the imbibed whole dicot seeds into the conical member. The imbibed whole dicot seeds can be added to the conical member by pouring them into the conical member. According to some embodiments, the imbibed whole dicot seeds can be stored on plate(s), and / or similar platform-type surface(s), prior to being added into the conical member. According to some embodiments, the imbibed whole dicot seeds can be added to the conical member one plate at a time. According to some embodiments the number of plates of imbibed whole dicot seeds can number from zero to thirteen, however any suitable number of plate(s) of whole dicot seeds could be included. When performing the step 532, it can be beneficial to avoid dropping any plate(s) into the conical member.

[0239] The method 510 further includes the step 534 comprising adding liquid to the conical member to create a suspension of the imbibed whole dicot sees and the liquid. According to some embodiments, enough of the liquid can be poured into the conical member and / or first container such that the liquid is filled to about 2 inches below where the whole dicot seeds sit in the conical member. However, the liquid could be filled to any suitable height. According to some embodiments, adding too much water lowers efficiency of embryonic extraction. According to some embodiments, adding too little water does not allow seeds to be mixed through the conical member.

[0240] The method 510 further includes the step 536 comprising allowing the suspension to rotate for an amount of time. According to various embodiments, the suspension can be allowed to rotate for an amount of time spanning from 35 to 105 minutes. According to someAgent Ref. No. P14771WOOO embodiments, the amount of time the suspension could be allowed to rotate is 50 minutes. According to some embodiments, the amount of time the suspension could be allowed to rotate is 35 minutes. During rotation, embryonic axes will be extracted from the whole dicot seeds and will pass through the at least one through-pores of the conical member wherein the passed embryonic axes with collect at or near the bottom and / or collection area of the first container.

[0241] The method 510 further includes the step 537 comprising, after allowing the suspension to rotate for an amount of time, adding further liquid to the suspension, adjusting the height of the impeller, and adjusting the speed of the rotary power source. According to some embodiments, the further liquid can be water, such as sterile water, however, any suitable liquid could be used. The amount of further liquid added to the suspension can vary according to various embodiments. The further liquid added to the suspension can be added to the top of the suspension. The adjustment of the height of the impeller can comprise adjusting the impeller so that the impeller body, which can comprise a brush and / or bristles, is at its lowest possible position inside the conical member. The adjustment of the speed of the rotary power source can comprise increasing the speed of the rotary power source to its highest setting and / or to a relatively high setting. This will cause passed embryonic axes to generally sink to the collection area of the first container and / or sink to the bottom or near the bottom of the first container. According to some embodiments, wherein the rotary power source is spun at 75-200 RPM for 35 minutes during the step 536, the step 537 can comprise adjusting the speed of the rotary power source to 250 RPM.

[0242] The method 510 further includes the step 538 comprising powering OFF the rotary power source and removing the impeller from the conical member. According to some embodiments, the impeller can be removed from the conical member by adjusting the height of the rotary power source such that the impeller is raised high enough wherein it is removed from the conical member and / or such that the height of the rotary power source is adjusted to its highest position. Additionally or alternatively, according to some embodiments, prior to removing the impeller from the conical member, the step 538 can comprise adjusting the height of the impeller such that the impeller body sits about 3 to 4 inches below the open top base end of the conical member. Then, when the impeller body is positioned about 3 to 4 inches below the open top base end of the conical member, the rotary power source can be powered ON to its lowest speed to allow any remaining seeds embryonic axes, and / or debris to spin out of any aspect(s) of the impeller such as the impeller body including any brush and / or bristles thereof. Additionally or alternatively, according to some embodiments, the step 538 can comprise removing and / or operatively detaching the impeller from the rotary power source.Agent Ref. No. P14771WOOO

[0243] The method 510 further includes the step 540 comprising removing the conical member from the first container. Such removal can comprise removing the conical member from the internal volume of the first container. Such removal can comprise unclamping the conical member from the first container. As stated, before removing the conical member from the first container, according to some embodiments, the rotary power source can be turned ON to its lowest speed, and / or a relatively low speed, to allow any remaining whole dicot seeds, extracted embryonic axes, and / or debris to spin out of any aspect(s) of the impeller such as the impeller body including any brush and / or bristles thereof. According to some embodiments, after allowing remaining whole dicot seeds, embryonic axes, and / or debris to spin out of any aspect(s) of the impeller, the rotary power source can be turned OFF again and the height of the rotary power source can be adjusted to its highest position, and / or a relatively high position. Further, according to some embodiments, the impeller can be removed from the rotary power source prior to unclamping the conical member and / or removing the conical member.

[0244] The method 510 further includes the step 542 comprising pouring out, and / or otherwise removing, excess liquid from the first container wherein at least a portion of debris is poured out, and / or otherwise removed, with the excess liquid but the passed embryonic axes generally remain in the first container. The debris can include any portions of the dicot seeds other than the embryonic axes including, but not limited to, seed coat(s), cotyledon(s), and the like. The excess liquid can be poured into any sort of waste receptacle and / or into any sort of drain and / or basin. According to some embodiments, the method 510 and / or any apparatus and / or system described herein can include zero or more waste receptacles to receive unwanted, excess, and / or used liquid and / or debris. According to some embodiments, a waste sieve can be used in conjunction with the removal of the excess water and / or debris. For example, the waste sieve can be added to and / or placed at or near the top of the waste receptacle(s), to catch and / or collect any embryonic axes that are poured into and / or onto the waste sieve. The waste sieve can be used similarly if pouring the excess water and / or debris into any sort of drain and / or basin rather than waste receptacle(s). The waste sieve can be the same as and / or similar to any sieve mentioned herein. For example, the waste sieve can be any type of sieve, strainer, filter, screen, mesh material, and the like capable of separating particulate matter having differing characteristics such as being of differing size. The waste sieve can be configured to allow debris and liquid to substantially pass through aperture(s) / pore(s) of the waste sieve while substantially catching and / or collecting embryonic axes. According to some embodiments, any embryonic axes caught by the waste sieve can be placed back into the first container and / or another container for collection. The waste receptacle(s) can be any sort of beaker, pitcher, cup, bucket,Agent Ref. No. P14771WOOO and the like. According to some embodiments, such waste receptacle(s) can be and / or comprise one or more Polypropylene Beakers with Handle manufactured by Carolina Biological Supply 580 (item number 721448) (see www.carolina.com / catalog / detail.jsp?prodId=721445&srsltid=AfmBOoqyLtbOKkoT92CQFtU8 JmYNWsm3n0SxHmeMVgqTlfXpwGlBytgLPaY (date accessed: October 18, 2024)). An example image of the Polypropylene Beaker(s) with Handle manufactured by Carolina Biological Supply 580 is shown in Figure 33. The Polypropylene Beaker(s) with Handle manufactured by Carolina Biological Supply 580 can be graduated in milliliters, can have a capacity of about 2,000 milliliters, and can include a handle.

[0245] The method 510 further includes the step 544 comprising adding an additional amount of liquid to the first container. According to some embodiments, the amount of additional liquid added to the first container can be in the range of tens to hundreds of milliliters. According to some embodiments, the amount of additional liquid added to the first container is about 500 milliliters. However, any suitable amount of additional liquid could be added to the first container. According to some embodiments the additional liquid is sterile water and / or any other type of water. However, any suitable type of liquid could be added to the first container as additional liquid.

[0246] The method 510 further includes the step 546 comprising moving the suspension / contents remaining in the first container wherein said suspension / remaining contents comprises at least a portion of the liquid, the additional amount of liquid, at least some of the passed embryonic axes, and another portion of the debris. Moving the remaining contents can comprise a user picking up the first container and moving the first container in a circular motion to suspend the embryonic axes. The first container can include a handle, according to some embodiments, wherein a user can pick up the first container using the handle and / or can use the handle to move the suspension / contents remaining in the first container. Additionally or alternatively, any suitable machine, device, and the like could be used to move the first container in a circular motion to suspend the embryonic axes. Additionally or alternatively, the remaining contents do not need to be moved in a circular motion. Rather, any suitable motion capable of properly suspending the embryonic axes could be employed.

[0247] The method 510 further includes the step 548 comprising pouring the remaining suspension in the first container into a second container. According to optional embodiments, a transition sieve can optionally be positioned relative to the second container wherein the remaining suspension in the first container is poured onto and / or into the transition sieve when pouring the contents of the first container into the second container. In such embodiments, theAgent Ref. No. P14771WOOO transition sieve can be the same as or similar to any sieve described herein such as the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430. The transition sieve can be positioned above, or otherwise relative to, the second container such that when the remaining suspension is poured onto / into the transition sieve, material that passes through the transition sieve enters the second container. This transition sieve is configured to substantially allow embryonic axes to pass into the second container while preventing most debris from passing into the second container. According to some embodiments, the step 548 can further comprise adding water to the transition sieve. According to some embodiments, the water can be sterile water. According to some embodiments, water, such as sterile water, can be added to fill the transition sieve up about 4 inches. However, according to various embodiments, any suitable amount of water could be added to the transition sieve. According to some embodiments wherein a transition sieve is used in conjunction with the second container when pouring the contents from the first container into the second container, the transition sieve can be moved in order to sieve the passed embryonic axes through the transition sieve, wherein the moving of the transition sieve continues until an amount of the passed embryonic axes are sieved through the transition sieve and into the second container. This amount of passed embryonic axes can vary. The moving of the transition sieve can continue until a majority of the passed embryonic axes are sieved through the transition sieve. The moving of the transition sieve can be performed by a user in an up-and-down motion. Additionally or alternatively, the moving of the transition sieve can be performed via a machine capable of moving the transition sieve. Additionally or alternatively, the moving of the transition sieve can be performed in a manner other than in an up-and-down motion such as a circular motion and / or any other suitable manner.

[0248] According to some embodiments, the second container can be a 2 liter container, such as the Polypropylene Beaker with Handle manufactured by Carolina Biological Supply 580 and / or the 2000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12A, both of which are described herein.

[0249] In certain embodiments, the method 510 further includes the step 552 comprising pouring out, and / or otherwise removing, an amount of liquid from the second container. According to various embodiments, this amount of liquid can vary. According to some embodiments, a majority of liquid is poured and / or otherwise removed from the second container. According to some embodiments wherein a transition sieve is optionally used when pouring the contents of the first container into the second container, the step 552 can further comprise removing the transition sieve from the second container prior to pouring out a majorityAgent Ref. No. P14771WOOO of the liquid from the second container. According to some embodiments, the liquid in the second container can be poured into the zero or more waste receptacles and / or into any sort of drain and / or basin.

[0250] According to some embodiments, the step 552 can optionally further comprise adding an amount of a liquid salt solution to the second container and allowing the second container to sit for an amount of time after adding the liquid salt solution. According to some embodiments, the liquid salt solution can be GM0046 liquid salt solution having a concentration of 6.8%. However, according to various embodiments, the liquid salt solution could be any other suitable liquid salt solution. For example, according to some embodiments, the liquid salt solution could be GM0049 liquid salt solution having a concentration of 3.4%. According to some embodiments, the liquid salt solution could be GM0045. According to some embodiments, the amount of liquid salt solution added to the second container is about 500 milliliters. However, any suitable amount of liquid salt solution can be added to the second container. According to some embodiments, the amount of time the mixture in the second container is allowed to sit is for about 20 minutes. However, the mixture in the second container can be allowed to sit for any suitable amount of time.

[0251] The method 510 further includes the step 556 comprising moving the second container to substantially separate the passed embryonic axes in the second container from debris in the second container. According to some embodiments, a user can move the second container in a circular motion. Additionally or alternatively, the second container can be moved in a circular motion by any suitable machine or device. Additionally or alternatively, the second container can be moved in any suitable manner capable of separating embryonic axes from debris. For example, according to some embodiments, a user could use a handle of the second container to move the second container in a circular motion and / or any other suitable motion. The moving of the second container is intended to separate the embryonic axes from debris by causing the embryonic axes to sink to the bottom of the second container while the debris floats. According to some embodiments, the second container can comprise a handle wherein a user and / or machine can use the handle in order to move the second container.

[0252] The method 510 further includes the step 558 comprising pouring out, and / or otherwise removing, at least a portion of the debris from the second container while at least a portion of the passed embryonic axes remain in the second container. Step 556 is configured to cause the passed embryonic axes to generally sink to the bottom of the second container and cause the debris in the second container to generally float. Thus, in step 558 a user, and / or machine, will be able to pour out a majority of the debris from the second container while a majority of theAgent Ref. No. P14771WOOO passed embryonic axes remain in the second container. According to some embodiments, the liquid and / or debris in the second container can be poured into the zero or more waste receptacle(s) and / or into any sort of drain and / or basin. Again, the waste sieve, and / or another waste sieve, can be used in conjunction with the waste receptacle(s) and / or in conjunction with pouring out the liquid and / or debris of the second container. This waste sieve can be the same as and / or similar to any sieve mentioned herein. For example, this waste sieve can be any type of sieve, strainer, filter, screen, mesh material, and the like capable of separating particulate matter having differing characteristics such as being of differing size. This waste sieve can be configured to substantially catch and / or collect any embryonic axes that are poured into and / or onto the waste sieve while allowing debris and liquid to substantially pass through the waste sieve into the waste receptacle(s) and / or into any drain and / or basin. According to some embodiments, any embryonic axes caught by such a waste sieve can be placed back into the second container and / or into any other container for collection. According to some embodiments, broken or damaged embryonic axes may float and may be poured out with the debris. Additionally or alternatively, according to some embodiments, the hilium of each of the dicot seeds may remain in the second container with the embryonic axes. If the hilium of one or more of the dicot seeds remain with the embryonic axes in the second container, any sort of use of the embryonic axes, such as a transformation, will not be effected. According to some embodiments, the step 558 should be performed quickly after the step 556 so that removal of at least a portion of the debris occurs while the passed embryonic axes and debris are generally separated, which makes removal of at least a portion of the debris easier without inadvertently removing passed embryonic axes.

[0253] According to some embodiments wherein a salt solution was added to the second container, the step 558 can further comprise removing an amount of the salt solution from the second container. According to various embodiments, this amount of salt solution can vary. According to some embodiments, a majority of the salt solution is removed from the second container. The salt solution can be removed by pouring and / or by any other suitable approach.

[0254] According to some embodiments wherein a salt solution was added to the second container, the step 558 can further comprise washing the second container with water. According to some embodiments, the water can be sterile water. According to some embodiments, the step 558 can comprise one or more washings of the second container. For example, according to some embodiments, the step 558 can comprise two washings of the second container.Agent Ref. No. P14771WOOO

[0255] The method 510 further includes the step 564 comprising resuspending the passed embryonic axes into solution in the second container. Throughout step 558, an amount of liquid can be left in the second container. For example, according to some embodiments, 100 to 200 milliliters of liquid can remain in the second container at the completion of step 558. Thus, step 564 comprises resuspending the passed embryonic axes in the liquid remaining in the second container at the completion of step 558. According to some embodiments, the liquid can be water and / or sterile water.

[0256] The method 510 further includes the step 566 comprising pouring at least a portion of the passed embryonic axes remaining in the second container into a third container. According to some embodiments, the third container can be a petri dish. Additionally or alternatively, according to some embodiments, the third container can be any sort of container and / or receptacle including, but not limited to, a pitcher, beaker, bowl, dish, pan, and the like. According to some embodiments, the third container could be any container described herein. According to some embodiments, the third container is and / or comprises the Nalgene Autoclavable Polypropylene Pan manufactured by Thermo Scientific 582 (catalog number 6902- 1000) (see www.fishersci.com / shop / products / nalgene-autoclavable-polypropylene-pans / p- 4520927 (date accessed: October 18, 2024)) (see www.thermofisher.com / order / catalog / product / 6902-1000 (date accessed: October 18, 2024)). An example image of the Nalgene Autoclavable Polypropylene Pan manufactured by Thermo Scientific 582 is shown in Figure 34. According to some embodiments, the Nalgene Autoclavable Polypropylene Pan manufactured by Thermo Scientific 582 can be autoclavable, can include heavy-duty construction (such as heavy wall construction which will withstand rugged use and repeated autoclaving), rolled edges to provide extra stiffness for carrying heavy contents, extra strong rims and rounded corners for easy cleaning, and can be tapered for nesting to save space. According to some embodiments, the Nalgene Autoclavable Polypropylene Pan manufactured by Thermo Scientific 582 can have the following dimensions: a capacity ranging from about 2 to 5 liters, a height ranging from about 64 to 107 millimeters, a top outside length ranging from about 260 to 264 millimeters, a top inside length ranging from about 236 to 300 millimeters, a bottom inside length ranging from about 203 to 267 millimeters, a top outside width ranging from about 159 to 325 millimeters, a top inside width ranging from about 135 to 239 millimeters, a bottom inside width ranging from about 102 to 208 millimeters, and a weight ranging from about 168 to 425 grams.

[0257] The method 510 further includes the step 568 comprising suspending the passed embryonic axes in a tissue culture medium. According to some embodiments, such suspensionAgent Ref. No. P14771WOOO involves the embryonic axes in the third container and such suspension can occur in the third container. According to some embodiments, such suspension is separate from the third container. A variety of tissue culture media are known that support plant tissue growth and development. These tissue culture media can either be purchased as a commercial preparation or custom prepared and modified by those of skill in the art. Examples of such tissue culture media include, but are not limited to, Murashige and Skoog (MS) medium, Linsmaier and Skoog (LS) medium, Gamborg (B5) medium, Nitsch and Nitsch (NN) medium, Chu (N6) medium, and Schenk and Hildebrandt medium, or derivations of these media. Those of skill in the art can also select various components such as basal salts, vitamins, and a carbon source from one or the other medium to obtain desired growth and development. According to some embodiments, the method 510 can further comprise using Agrobacterium to infect the embryonic axes collected in the third container. This could include suspending the embryonic axes in the third container in Agrobacterium. For example, according to some embodiments, Agrobacterium co-culture (CC) medium could be used. Additionally or alternatively, according to some embodiments, the infection of embryonic axes with Agrobacterium can be performed using protocols used to infect soybean embryonic axes obtained by manual techniques. According to some embodiments, the collected embryonic axes can be used for a biolistic transformation, biolistic gene delivery, an agrobacterial and / or Agrobacterium-mediated transformation, and / or gene editing. According to some embodiments, the passed embryonic axes in the third container could be suspended in and / or exposed to a tissue culture medium comprising biolistic pre-bombardment medium in which embryos can also be incubated for biolistic transformation as well as other applications. Additionally or alternatively, according to some embodiments, the step 568 can comprise infecting explants with Agrobacterium using embryonic axis protocol.

[0258] It should be noted that, according to some embodiments, any steps of the method 510 can be repeated, if desired, to continue to substantially separate / extract embryonic axes from any whole seeds that remain and to collect the substantially separated / extracted embryonic axes. It should also be noted that, according to various embodiments, any step of the method 510 can be optional and that any step of the method 510 can be omitted and / or performed in a different sequence than what is shown in Figures 29A-E.

[0259] Methods of introducing exogenous nucleic acids into plant cells are well known in the art. As used herein, the term “introduced” means providing a nucleic acid (e.g., expression construct) or protein into a cell. “Introduced” includes reference to the incorporation of a nucleic acid into a eukaryotic or prokaryotic cell where the nucleic acid may be incorporated into the genome of the cell and includes reference to the transient provision of a nucleic acid or proteinAgent Ref. No. P14771WOOO to the cell. “Introduced” includes reference to stable or transient transformation methods. Thus, “introduced” in the context of inserting a nucleic acid fragment (e.g., a recombinant DNA construct / expression construct) into a cell, means “transfection” or “transformation” or “transduction” and includes reference to the incorporation of a nucleic acid fragment into a eukaryotic or prokaryotic cell where the nucleic acid fragment may be incorporated into the genome of the cell (e.g., nuclear chromosome, plasmid, plastid, chloroplast, or mitochondrial DNA), converted into an autonomous replicon, or transiently expressed (e.g., transfected mRNA).

[0260] Suitable methods include viral infection (such as double stranded DNA viruses including geminiviruses), transfection, conjugation, protoplast fusion, electroporation, particle gun technology, calcium phosphate precipitation, direct microinjection, silicon carbide whiskers technology, Agrobacterium-mediated transformation and the like. The choice of method is generally dependent on the type of cell being transformed and the circumstances under which the transformation is taking place (e.g., in vitro, ex vivo, or in vivo).

[0261] Bacterially-mediated transformation methods based upon the soil bacterium Agrobacterium tumefaciens or other bacteria (e.g., Rhizobium sp., Sinorhizobium sp., Mesorhizobium sp., Bradyrhizobium sp., Azobacter sp., Phyllobacterium sp. see, e.g., Broothaerts et al. (2005) Nature, 433:629-633) are particularly useful for introducing an exogenous nucleic acid molecule into a vascular plant. The wild type form of Agrobacterium contains a Ti (tumor-inducing) plasmid that directs production of tumorigenic crown gall growth on host plants. Transfer of the tumor-inducing T-DNA region of the Ti plasmid to a plant genome requires the Ti plasmid-encoded virulence genes as well as T-DNA borders, which are a set of direct DNA repeats that delineate the region to be transferred. An Agrobacterium-based vector is a modified form of a Ti plasmid, in which the tumor inducing functions are replaced by the nucleic acid sequence of interest to be introduced into the plant host.

[0262] Agrobacterium-mediated transformation generally employs cointegrate vectors or binary vector systems, in which the components of the Ti plasmid are divided between a helper vector, which resides permanently in the Agrobacterium host and carries the virulence genes, and a shuttle vector, which contains the gene of interest bounded by T-DNA sequences. A variety of binary vectors are well known in the art and are commercially available, for example, from Clontech (Palo Alto, Calif.). Methods of coculturing Agrobacterium with cultured plant cells or wounded tissue such as leaf tissue, root explants, hypocotyls, cotyledons, stem pieces or tubers, for example, also are well known in the art. See, e.g., Glick and Thompson, (eds.), Methods in Plant Molecular Biology and Biotechnology, Boca Raton, Fla.: CRC Press (1993).Agent Ref. No. P14771WOOO

[0263] Microprojectile-mediated transformation also can be used to transform a plant cell. This method, first described by Klein et al. (Nature 327:70-73 (1987)), relies on microprojectiles such as gold or tungsten that are coated with the desired nucleic acid molecule by precipitation with calcium chloride, spermidine or polyethylene glycol. The microprojectile particles are accelerated at high speed into an angiosperm tissue using a device such as the BIOLISTIC PD- 1000 (Biorad; Hercules Calif.). A nucleic acid may be introduced into a plant in a manner such that the nucleic acid is able to enter a plant cell(s), e.g., via an in vivo or ex vivo protocol. By "in vivo," it is meant in the nucleic acid is administered to a living body of a plant e.g., infiltration. By "ex vivo" it is meant that cells or explants are modified outside of the plant, and then such cells or organs are regenerated to a plant. A number of vectors suitable for stable transformation of plant cells have been described, including those described in Weissbach and Weissbach, (1989) Methods for Plant Molecular Biology Academic Press, and Gelvin et al., (1990) Plant Molecular Biology Manual, Kluwer Academic Publishers. Specific examples include those derived from a Ti plasmid of Agrobacterium tumefaciens, as well as those disclosed by Herrera- Estrella et al. (1983) Nature 303: 209, Bevan (1984) Nucl Acid Res. 12: 8711-8721, Klee (1985) Bio / Technolo 3: 637-642.

[0264] Alternatively, non- Ti vectors can be used to transfer the DNA into plants and cells by using free DNA delivery techniques. By using these methods transgenic plants such as wheat, rice (Christou (1991) Bio / Technology 9:957-9 and 4462) and com (Gordon-Kamm (1990) Plant Cell 2: 603-618) can be produced. An immature embryo can also be a good target tissue for monocots for direct DNA delivery techniques by using the particle gun (Weeks et al. (1993) Plant Physiol 102: 1077-1084; Vasil (1993) Bio / Technolo 10: 667-674; Wan and Lemeaux (1994) Plant Physiol 104: 37-48 and for Agrobacterium-mediated DNA transfer (Ishida et al. (1996) Nature Biotech 14: 745-750). Methods for introduction of DNA into chloroplasts are biolistic bombardment, polyethylene glycol transformation of protoplasts, and microinjection (Danieli et al Nat. Biotechnol 16:345-348, 1998; Staub et al Nat. Biotechnol 18: 333-338, 2000; O'Neill et al Plant J. 3:729-738, 1993; Knoblauch et al Nat. Biotechnol 17: 906-909; U.S. Pat. Nos. 5,451,513, 5,545,817, 5,545,818, and 5,576,198; in Inti. Application No. WO 95 / 16783; and in Boynton et al., Methods in Enzymology 217: 510-536 (1993), Svab et al., Proc. Natl. Acad. Sci. USA 90: 913-917 (1993), and McBride et al., Proc. Natl. Acad. Sci. USA 91 : 7301- 7305 (1994)). Any vector suitable for the methods of biolistic bombardment, polyethylene glycol transformation of protoplasts and microinjection will be suitable as a targeting vector for chloroplast transformation. Any double stranded DNA vector may be used as a transformation vector, especially when the method of introduction does not utilize Agrobacterium.Agent Ref. No. P14771WOOO

[0265] Gene editing molecules of use in methods and systems provided herein include molecules capable of introducing a double-strand break (“DSB”) or single-strand break (“SSB”) at a specific site or sequence in a double-stranded DNA, such as in genomic DNA or in a target gene located within the genomic DNA as well as accompanying guide RNA. Examples of such gene editing molecules include: (a) a nuclease comprising an RNA-guided nuclease, an RNA- guided DNA endonuclease or RNA directed DNA endonuclease (RdDe), a class 1 CRISPR type nuclease system, a class 2 type II Cas nuclease, a Cas9, a nCas9 nickase, a class 2 type V Cas nuclease, a Cas 12a nuclease, a nCasl2a nickase, a Cas 12d (CasY), a Casl2e (CasX), a Cas 12b (C2cl), a Casl2c (C2c3), a Casl2i, a Casl2j, a Casl4, an engineered nuclease, a codon- optimized nuclease, a zinc-finger nuclease (ZFN) or nickase, a transcription activator-like effector nuclease (TAL-effector nuclease or TALEN) or nickase (TALE-nickase), an Argonaute, and a meganuclease or engineered meganuclease; (b) a polynucleotide encoding one or more nucleases capable of effectuating site-specific alteration (including introduction of a DSB or SSB) of a target nucleotide sequence; (c) a guide RNA (gRNA) for use with an RNA-guided nuclease, or a DNA encoding a gRNA for use with an RNA-guided nuclease; (d) optionally donor DNA template polynucleotides suitable for insertion at a break in genomic DNA by homology-directed repair (HDR) or microhomology-mediated end joining (MMEJ); and (e) optionally other DNA templates (e.g., dsDNA, ssDNA, or combinations thereof) suitable for insertion at a break in genomic DNA (e.g., by non-homologous end joining (NHEJ). In certain embodiments, the at least one mutation is made with a cytosine and / or adenine base editor, or by a PRIME editing system.

[0266] CRISPR technology for editing the genes of eukaryotes is disclosed in US Patent Application Publications 2016 / 0138008 Al and US2015 / 0344912A1, and in US Patents 8,697,359, 8,771,945, 8,945,839, 8,999,641, 8,993,233, 8,895,308, 8,865,406, 8,889,418, 8,871 ,445, 8,889,356, 8,932,814, 8,795,965, and 8,906,616. Cpfl endonuclease and corresponding guide RNAs and PAM sites are disclosed in US Patent Application Publication 2016 / 0208243 Al . Plant RNA promoters for expressing CRISPR guide RNA and plant codon- optimized CRISPR Cas9 endonuclease are disclosed in International Patent Application PCT / US2015 / 018104 (published as WO 2015 / 131 101 and claiming priority to US Provisional Patent Application 61 / 945,700). Methods of using CRISPR technology for genome editing in plants are disclosed in US Patent Application Publications US 2015 / 0082478A1 and US 2015 / 0059010A1 and in International Patent Application PCT / US2015 / 038767 Al (published as WO 2016 / 007347 and claiming priority to US Provisional Patent Application 62 / 023,246). All of the patent publications referenced in this paragraph are incorporated herein by reference inAgent Ref. No. P14771WOOO their entirety. In certain embodiments, an RNA-guided endonuclease that leaves a blunt end following cleavage of the target site is used. Blunt-end cutting RNA-guided endonucleases include Cas9. In certain embodiments, an RNA-guided endonuclease that leaves a staggered single stranded DNA overhanging end following cleavage of the target site following cleavage of the target site is used. Staggered-end cutting RNA-guided endonucleases include Casl2a, Cast 2b, Cast 2d, Casl2e, and Casl2i.

[0267] CRISPR-type genome editing can be adapted for use in the methods and systems provided herein in several ways. CRISPR elements, e.g., gene editing molecules comprising CRISPR endonucleases and CRISPR guide RNAs including single guide RNAs or guide RNAs in combination with tracrRNAs or scoutRNA, or polynucleotides encoding the same, are useful in effectuating genome editing without remnants of the CRISPR elements or selective genetic markers occurring in progeny. In certain embodiments, the CRISPR elements are provided directly to the dicot cell (e.g., soybean plant cells), systems, methods, and compositions as isolated molecules, as isolated or semi-purified products of a cell free synthetic process (e.g., in vitro translation), or as isolated or semi-purified products of in a cell-based synthetic process (e.g., such as in a bacterial or other cell lysate). In certain embodiments, dicot plants or dicot plant cells used in the systems, methods, and compositions provided herein can comprise a transgene that expresses a CRISPR endonuclease (e.g., a Cas9, a Cpfl-type or other CRISPR endonuclease). In certain embodiments, one or more CRISPR endonucleases with unique PAM recognition sites can be used. Guide RNAs (sgRNAs or crRNAs and a tracrRNA or scoutRNA) to form an RNA-guided endonuclease / guide RNA complex which can specifically bind sequences in the gDNA target site that are adjacent to a protospacer adjacent motif (PAM) sequence. The type of RNA-guided endonuclease typically informs the location of suitable PAM sites and design of crRNAs or sgRNAs. G-rich PAM sites, e.g., 5’-NGG are typically targeted for design of crRNAs or sgRNAs used with Cas9 proteins. Examples of PAM sequences include 5’-NGG (Streptococcus pyogenes), 5’-NNAGAA (Streptococcus thermophilus CRISPR1), 5’- NGGNG (Streptococcus thermophilus CRISPR3), 5’-NNGRRT or 5’-NNGRR (Staphylococcus aureus Cas9, SaCas9), and 5’-NNNGATT (Neisseria meningitidis). T-rich PAM sites (e.g., 5’- TTN or 5’-TTTV, where “V” is A, C, or G) are typically targeted for design of crRNAs or sgRNAs used with Casl2a proteins. In some instances, Casl2a can also recognize a 5’-CTA PAM motif. Other examples of potential Casl2a PAM sequences include TTN, CTN, TCN, CCN, TTTN, TCTN, TTCN, CTTN, ATTN, TCCN, TTGN, GTTN, CCCN, CCTN, TTAN, TCGN, CTCN, ACTN, GCTN, TCAN, GCCN, and CCGN (wherein N is defined as any nucleotide). Cpfl endonuclease and corresponding guide RNAs and PAM sites are disclosed inAgent Ref. No. P14771WOOOUS Patent Application Publication 2016 / 0208243 Al, which is incorporated herein by reference for its disclosure of DNA encoding Cpfl endonucleases and guide RNAs and PAM sites. Engineered endonucleases with altered or eliminated PAM recognition sites can also be used.

[0268] Zinc-finger nucleases are site-specific endonucleases comprising two protein domains: a DNA-binding domain, comprising a plurality of individual zinc finger repeats that each recognize between 9 and 18 base pairs, and a DNA-cleavage domain that comprises a nuclease domain (typically Fokl). The cleavage domain dimerizes in order to cleave DNA; therefore, a pair of ZFNs are required to target non-palindromic target polynucleotides. In certain embodiments, zinc finger nuclease and zinc finger nickase design methods which have been described (Umov et al. (2010) Nature Rev. Genet., 11 :636 - 646; Mohanta et al. (2017) Genes vol. 8,12: 399; Ramirez et al. Nucleic Acids Res. (2012); 40(12): 5560-5568; Liu et al. (2013) Nature Communications, 4: 2565) can be adapted for use in the methods set forth herein. The zinc finger binding domains of the zinc finger nuclease or nickase provide specificity and can be engineered to specifically recognize any desired target DNA sequence. The zinc finger DNA binding domains are derived from the DNA-binding domain of a large class of eukaryotic transcription factors called zinc finger proteins (ZFPs). The DNA-binding domain of ZFPs typically contains a tandem array of at least three zinc “fingers” each recognizing a specific triplet of DNA. A number of strategies can be used to design the binding specificity of the zinc finger binding domain. One approach, termed “modular assembly”, relies on the functional autonomy of individual zinc fingers with DNA. In this approach, a given sequence is targeted by identifying zinc fingers for each component triplet in the sequence and linking them into a multifmger peptide. Several alternative strategies for designing zinc finger DNA binding domains have also been developed. These methods are designed to accommodate the ability of zinc fingers to contact neighboring fingers as well as nucleotide bases outside their target triplet. Typically, the engineered zinc finger DNA binding domain has a novel binding specificity, compared to a naturally occurring zinc finger protein. Engineering methods include, for example, rational design and various types of selection. Rational design includes, for example, the use of databases of triplet (or quadruplet) nucleotide sequences and individual zinc finger amino acid sequences, in which each triplet or quadruplet nucleotide sequence is associated with one or more amino acid sequences of zinc fingers which bind the particular triplet or quadruplet sequence. See, e.g., US Patents 6,453,242 and 6,534,261, both incorporated herein by reference in their entirety. Exemplary selection methods (e.g., phage display and yeast two-hybrid systems) can be adapted for use in the methods described herein. In addition, enhancement of binding specificity for zinc finger binding domains has been described in US Patent 6,794,136,Agent Ref. No. P14771WOOO incorporated herein by reference in its entirety. In addition, individual zinc finger domains may be linked together using any suitable linker sequences. Examples of linker sequences are publicly known, e.g., see US Patents 6,479,626; 6,903,185; and 7,153,949, incorporated herein by reference in their entirety. The nucleic acid cleavage domain is non-specific and is typically a restriction endonuclease, such as Fokl. This endonuclease must dimerize to cleave DNA. Thus, cleavage by Fokl as part of a ZFN requires two adjacent and independent binding events, which must occur in both the correct orientation and with appropriate spacing to permit dimer formation. The requirement for two DNA binding events enables more specific targeting of long and potentially unique recognition sites. Fokl variants with enhanced activities have been described and can be adapted for use in the methods described herein; see, e.g., Guo et al. (2010) J. Mol. Biol., 400:96 - 107.

[0269] Transcription activator like effectors (TALEs) are proteins secreted by certain Xanthomonas species to modulate gene expression in host plants and to facilitate the colonization by and survival of the bacterium. TALEs act as transcription factors and modulate expression of resistance genes in the plants. Recent studies of TALEs have revealed the code linking the repetitive region of TALEs with their target DNA-binding sites. TALEs comprise a highly conserved and repetitive region consisting of tandem repeats of mostly 33 or 34 amino acid segments. The repeat monomers differ from each other mainly at amino acid positions 12 and 13. A strong correlation between unique pairs of amino acids at positions 12 and 13 and the corresponding nucleotide in the TALE-binding site has been found. The simple relationship between amino acid sequence and DNA recognition of the TALE binding domain allows for the design of DNA binding domains of any desired specificity. TALEs can be linked to a nonspecific DNA cleavage domain to prepare genome editing proteins, referred to as TAL-effector nucleases or TALENs. As in the case of ZFNs, a restriction endonuclease, such as Fokl, can be conveniently used. Methods for use of TALENs in plants have been described and can be adapted for use in the methods described herein, see Mahfouz et al. (2011) Proc. Natl. Acad. Sci. USA, 108:2623 - 2628; Mahfouz (2011) GM Crops, 2:99 - 103; and Mohanta et al. (2017) Genes vol. 8,12: 399). TALE nickases have also been described and can be adapted for use in methods described herein (Wu et al.; Biochem Biophys Res Commun. (2014);446(1):261 -6; Luo et al; Scientific Reports 6, Article number: 20657 (2016)).

[0270] Figure 30 shows a system 610 for extracting / separating and collecting embryonic axes from whole dicot seeds. The system 610 can comprise an embryonic axis extraction and collection apparatus 612, a second container 616, a third container 618, and a tissue culture medium 620. The system 610 can operate according to any aspect(s) of any of the methodsAgent Ref. No. P14771WOOO described herein including any aspect(s) of the method 310, the method 410, and / or the method 510

[0271] The apparatus 612 can be and / or comprise any embryonic axis extraction and collection apparatus described herein such as the apparatus 10, the apparatus 110, the apparatus 1210, and / or the apparatus 1310. The apparatus 612 can function to substantially extract / separate and collect embryonic axes from the whole dicot seeds in substantially the same and / or similar manner as described herein regarding the apparatus 10, the apparatus 110, the apparatus 1210, and / or the apparatus 1310. For example, component s) of the apparatus 10, the apparatus 110, the apparatus 1210, and / or the apparatus 1310 can be used for rotating a suspension comprising a set of imbibed whole dicot seeds and a liquid relative to a surface having through-pores wherein the surface is effective to: (1) comminute the set of imbibed whole dicot seeds into smaller pieces, and (2) substantially pass embryonic axes through the through-pores. Further, component s) of the apparatus 10, the apparatus 110, the apparatus 1210, and / or the apparatus 1310 can be used for collecting the passed embryonic axes.

[0272] According to some embodiments, the system 610 can optionally include a transition sieve 614 as described herein. According to some embodiments, the transition sieve 614 can be the Tenta Kitchen Micro-Perforated Stainless Steel Pasta Basket manufactured by TENTA7 430. Figure 31 shows an isolated perspective view of an example embodiment of the transition sieve 614 positioned, at least partially, within the second container 616. As shown in Figure 31, the transition sieve 614 can comprise a strainer portion 622 wherein the strainer portion 622 includes one or more apertures 624. The strainer portion 622 can be generally cylindrical such that liquid and / or particulate matter can be poured into / onto the strainer portion 622. As shown in Figure 31, the transition sieve 614 can further comprise a handle 626. The handle 626 can extend generally from a point along the perimeter of the strainer portion 622. The diameter of each of the one or more apertures 624 can be properly sized such that the one or more apertures 624 are effective to substantially pass substantially intact embryonic axes. According to some embodiments, the diameter of each of the aperture(s) 624 can be approximately 6 millimeters. However, any suitable size of apertures can be included in the strainer portion 622 of the transition sieve 614. The transition sieve 614 can be adapted to include aperture(s) that are each about 6 millimeters in diameter. For example, for some embodiments, aperture(s) having a diameter of 6 millimeters can be drilled and / or cut into the side(s) and / or bottom of the transition sieve 614

[0273] After embryonic axes are extracted from the whole dicot seeds using the extraction and collection apparatus 612, passed through the through-pores, and collected in the container of theAgent Ref. No. P14771WOOO embryonic axis extraction and collection apparatus 612 (the first container), according to some embodiments, a suspension comprising liquid, embryonic axes passed through the through-pores of the conical member, and debris remains in the collection area of the first container. This remaining suspension comprising liquid, passed embryonic axes, and debris can be poured into the second container 616 such that the passed embryonic axes collect in the second container 616. Optionally, according to some embodiments, the transition sieve 614 can be used when pouring the contents of the first container of the apparatus 612 into the second container 616. The transition sieve 614 can be positioned relative to the second container 616, such as above and / or at least partially within the second container 616, such that when the contents of the first container of the apparatus 612 are poured into / onto the transition sieve 614, the transition sieve 614 substantially allows embryonic axes to pass through the transition sieve 614 into the second container 616 and substantially stops and / or catches debris, preventing such debris from passing into the second container 616. In this way, the transition sieve 614 is configured to further isolate embryonic axes from debris. Thus, the second container 616, can receive and collect passed embryonic axes, the liquid (such as water and / or sterile water), and any debris that inadvertently passed through the conical member of the apparatus 612, and / or, for embodiments that include the transition sieve 614, any debris that inadvertently passed through the transition sieve 614. According to some embodiments wherein no transition sieve is used, the contents of the first container of the apparatus 612 can be poured directly into the second container 616 without using a sieve.

[0274] According to some embodiments that use the transition sieve 614, in order to filter the embryonic axes through the transition sieve 614 to collect in the second container 616, a user and / or any sort of machine and / or device can move the transition sieve 614. For example, the transition sieve 614 can be moved in an up-and-down motion in order to pass the embryonic axes through the transition sieve 614. Additionally or alternatively, the transition sieve 614 can be moved in any other type of motion such as a circular motion. Movement of the transition sieve 614 can continue until an amount of embryonic axes have passed through the transition sieve. For example, according to some embodiments, movement of the transition sieve 614 can continue until a majority of the embryonic axes poured into / onto the transition sieve 614 pass through the transition sieve 614.

[0275] The second container 616 can be the same as and / or similar to any container described herein such as the container 12, the container 112, the container 1212, and / or the container 1312. For example, the second container 616 can be any sort of container and / or receptacle capable of holding liquid and particulate matter. For example, the second container 616 can be any off-the-Agent Ref. No. P14771WOOO shelf beaker, pitcher, bowl, pan, dish, and the like. The second container 616 can be a 2 liter pitcher / beaker, such as the Polypropylene Beaker with Handle manufactured by Carolina Biological Supply 580 and / or the 2000 milliliter Polypropylene Graduated Pitcher manufactured by Bel-Art 12A, both of which are described herein.

[0276] The second container 616 is configured to receive, catch, and / or otherwise collect the passed embryonic axes from the first container. The second container 616 is configured such that it has an open top or other opening wherein liquid and / or other materials can enter and / or exit the second container 616. Thus, passed embryonic axes, liquid, and any inadvertently passed debris can enter the second container 616 through the open top and / or any other opening wherein the passed embryonic axes, liquid, and any debris is collected in the second container 616. It is appreciated that the second container 616 can include, according to some embodiments, a lid and / or any sort of cover capable of closing and / or covering the open top and / or other opening of the second container 616.

[0277] As noted herein, the open top and / or other opening of the second container 616 is configured such that liquid and / or other material(s) can enter and / or exit the second container 616. For example, liquid in the second container 616 can be poured out of the second container 616 and / or otherwise removed from the second container 616 in any suitable manner. Additionally, liquid and / or other material(s), such as a liquid salt solution, can be poured into the second container 616 and / or otherwise enter the second container 616 in any suitable manner.

[0278] The second container 616 can be further configured such that it can be moved, by a user and / or any sort of machine and / or device, such that passed embryonic axes are substantially separated from any debris in the second container 616. Such movement of the second container 616 can be in a circular motion and / or in any other suitable motion.

[0279] As noted, the system 610 can further include a third container 618. The third container 618 can be configured to receive the passed embryonic axes from the second container 616. The third container 618 can be the same as and / or similar to any container described herein such as the container 12, the container 112, the container 1212, and / or the container 1312. For example, the third container 618 can be any sort of container and / or receptacle capable of holding liquid and particulate matter. For example, the third container 618 can be any off-the-shelf beaker, pitcher, bowl, pan, dish, and the like. According to some embodiments, the third container 618 can be and / or comprise the Nalgene Autoclavable Polypropylene Pan manufactured by Thermo Scientific 582. The third container 618 can include an open top and / or any other kind of opening wherein liquid and / or other material(s) can enter and / or exit the third container 618. For example, passed embryonic axes can be poured from the second container 616 into the thirdAgent Ref. No. P14771WOOO container 618 wherein the passed embryonic axes enter the third container 618 via the open top and / or another opening. It is appreciated that the third container 618 can include, according to some embodiments, a lid and / or any sort of cover capable of closing and / or covering the open top and / or other opening of the third container 618. According to some embodiments, the third container 618 is and / or comprises a petri dish.

[0280] As noted, the system 610 can further include a tissue culture medium 620. The tissue culture medium 620 can be configured to receive passed embryonic axes from the second container 616 and / or from the third container 618. While the tissue culture medium 620 is shown to be separate from the third container 618 in Figure 30, according to some embodiments, the tissue culture medium 620 can be positioned, at least partially, within the third container 618. The tissue culture medium 620 is configured to be capable of suspending one or more embryonic axes such as the passed embryonic axes. The tissue culture medium 620 can be and / or comprise any known tissue culture medium.

[0281] As shown in Figure 30, according to some embodiments, the system 610 can include a waste sieve 621. The waste sieve 621 can be and / or comprise any sort of sieve, filter, screen, mesh material, surface with aperture(s) / pore(s), and the like capable of separating particular matter having different characteristics such as having different sizes. For example, the waste sieve 621 can be the same as and / or similar to any sieve mentioned herein. According to some embodiments, the waste sieve 621 can be used in conjunction with one or more waste receptacle(s) 634, and / or other any sort of drain and / or basin, which can also be included as part of the system 610. For example, the waste sieve 621 can be added to and / or placed at or near the top of the waste receptacle(s) 634, and / or any sort of drain and / or basin, wherein, when contents of the first, second, or third container are poured into the waste receptacle(s) 634, and / or any sort of drain and / or basin, to remove liquid and / or debris, the contents are first poured into / onto the waste sieve 621 wherein the waste sieve 621 can catch and / or collect any embryonic axes that are poured into / onto the waste sieve 621. The waste sieve 621 can be configured to allow debris and / or liquid to substantially pass through aperture(s) / pore(s) of the waste sieve 621 while substantially catching and / or collecting embryonic axes. According to some embodiments, any embryonic axes caught by the waste sieve 621 can be placed into the first container of the system 610, the second container 616, the third container 618, and / or any other container and / or receptacle.

[0282] Therefore, as understood from the disclosure, the apparatus(es), method(s), and system(s) disclosed herein provide the ability to automatically, or at least semi-automatically, extract / separate embryonic axes from whole dicot seeds and to collect the embryonic axes. TheAgent Ref. No. P14771WOOO disclosure provides that such automated, or at least semi-automated, extract! on / separati on and collection of embryonic axes is performed in a fast, efficient, effective, and cost-effective manner. For example, the apparatus(es), method(s) and system(s) disclosed herein are cost- effective, in that they are configured to automatically, or at least semi-automatically extract embryonic axes from seeds in a much faster and more efficient manner than manual extraction. The apparatus(es), method(s), and system(s) disclosed herein greatly reduce time and effort spent extracting and collecting embryonic axes. This reduces costs associated with paying researchers and / or paying for laboratory space and / or time. The disclosed apparatus(es), method(s), and / or system(s) also promote innovation as the collected embryonic axes can be used in a variety of different ways including, but not limited to, a biolistic transformation, biolistic gene delivery, an agrobacterial transformation, and / or gene editing.Embodiments

[0283] Various embodiments of the systems, methods, and apparatus(es) described herein are set forth in the following set of numbered embodiments.

[0284] 1. A method for automated separation and collection of embryonic axes of dicot seeds, the method comprising: rotating a suspension of a plurality of whole dicot seeds and a liquid relative to a surface having through-pores, the surface being effective to: comminute the whole dicot seeds into smaller pieces, and substantially pass embryonic axes; and collecting the passed embryonic axes.

[0285] 2. The method of embodiment 1, wherein the plurality of whole dicot seeds comprises a plurality of seeds of the Fabaceae family.

[0286] 3. The method of embodiments 1 or 2, wherein the comminution imposes on the whole dicot seeds one or more of: friction; shear or impact stress(es); and / or degradation.

[0287] 4. The method of any one of embodiments 1-3, further comprising, while rotating the suspension, influencing the seeds toward or to the surface.

[0288] 5. The method of any one of embodiments 1-4, wherein the influencing comprises one or more of: centrifugal force(s); mechanical force(s); and / or gravitational force(s).

[0289] 6. The method of any one of embodiments 1-5, wherein the surface comprises: an internal portion of an inverted three-dimensional conical member positioned in an internal volume of a container, the conical member having an internal space along an axis between an open top base end and a closed opposite bottom end; and wherein the internal portion is between the open top base end and the closed opposite bottom end.Agent Ref. No. P14771WOOO

[0290] 7. The method of embodiment 6, wherein the rotating comprises rotating the suspension with an impeller: operably connected to a rotary power source; and wherein the impeller comprises an impeller body having top and bottom ends along a rotational axis, wherein the impeller body is adjustable relative to the closed opposite bottom end of the conical member.

[0291] 8. The method of embodiment 7, wherein the impeller body comprises: a structure extending towards or to the internal portion of the conical member when in operative position; bristles extending generally laterally from the rotational axis towards, at, or near the internal portion of the conical member when in operative position; or one or more flanges extending generally laterally from the rotational axis towards, at, or near the internal portion of the conical member when in operative position.

[0292] 9. The method of embodiment 8, further comprising: autoclaving the impeller body prior to the rotation of the suspension; and sterilizing the impeller body prior to the rotation of the suspension.

[0293] 10. The method of embodiments 8 or 9, further comprising adjusting height of the bottom end of the impeller body such that the bottom end of the impeller body is positioned within several inches from the closed opposite bottom end of the conical member.

[0294] 11. The method of any one of embodiments 6-10, further comprising securing the conical member to the container via a clamp prior to the rotating of the suspension.

[0295] 12. The method of any one of embodiments 6-11, wherein the collecting of the passed embryonic axes further comprises at least one of the following steps: removing the conical member from the internal volume of the container and pouring out excess liquid from the container wherein at least a portion of debris is poured out with the excess liquid but the passed embryonic axes generally remain in the container; adding an additional amount of liquid to the container; moving a suspension remaining in the container wherein said remaining suspension comprises at least a portion of the liquid, the additional amount of liquid, the passed embryonic axes, and another portion of the debris; pouring the remaining suspension into a second container; pouring out a majority of liquid from the second container; moving the second container to separate the passed embryonic axes in the second container from debris in the second container; pouring out the debris from the second container while the passed embryonic axes remain in the second container; resuspending the passed embryonic axes in the second container; and / or pouring the passed embryonic axes into a third container.

[0296] 13. The method of embodiment 12, wherein the additional amount of liquid is 500 milliliters.

[0297] 14. The method of embodiments 12 or 13, wherein the third container is a petri dish.Agent Ref. No. P14771WOOO

[0298] 15. The method of any one of embodiments 12-14, further comprising suspending the passed embryonic axes in a tissue culture medium.

[0299] 16. The method of any one of embodiments 1-15, wherein: the plurality of whole dicot seeds are soybean seeds; a pore size of each of the through-pores is on the order of or 6 millimeters; and the suspension is rotated on the order of 75 to 200 or 300 RPM.

[0300] 17. The method of any one of embodiments 1-16, wherein the collecting of the passed embryonic axes is for the purpose of one or more of: a biolistic transformation; biolistic gene delivery; an agrobacterial transformation; and / or gene editing.

[0301] 18. The method of any one of embodiments 1-17, wherein the whole dicot seeds are imbibed for several hours prior to being rotated in the suspension.

[0302] 19. The method of any one of embodiments 1-18, wherein the liquid is sterile water.

[0303] 20. An apparatus for automated separation and collection of embryonic axes of dicot seeds, the apparatus comprising: a container having: an internal volume configured to receive and retain a suspension of a set of imbibed whole dicot seeds and a liquid; an inverted three- dimensional conical member positioned at least partially in the internal volume of the container, the conical member having: an internal space along an axis between an open top base end and a closed opposite bottom end; and an internal surface between the open top base end and the closed opposite bottom end with through-pores, the internal surface being effective to: comminute the imbibed whole dicot seeds; and substantially pass embryonic axes of each of the imbibed whole dicot seeds; an impeller having: an interface configured to operably connect to a rotary power source; and an impeller body extending from a base end to an opposite end along a rotational axis, the impeller body configured to: rotate the suspension; and influence the set of imbibed whole dicot seeds toward or to the internal surface of the conical member; and a collection area in the container outside the internal space of the conical member to collect embryonic axes that have passed through the through-pores.

[0304] 21. The apparatus of embodiment 20, wherein the conical member is a circular conical member.

[0305] 22. The apparatus of embodiments 20 or 21 , wherein the conical member is a truncated cone with the closed opposite bottom end closed at a truncation plane.

[0306] 23. The apparatus of any one of embodiments 20-22, wherein the internal surface and the impeller together are configured to degrade and / or comminute the set of imbibed whole dicot seeds.

[0307] 24. The apparatus of any one of embodiments 20-23, wherein a pore size of each of the through-pores is on the order of or 6 millimeters.Agent Ref. No. P14771WOOO

[0308] 25. The apparatus of any one of embodiments 20-24, wherein the impeller body has a cylindrical shape, conical shape, substantially cylindrical shape, or substantially conical shape.

[0309] 26. The apparatus of any one of embodiments 20-25, wherein the impeller body comprises bristles extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.

[0310] 27. The apparatus of embodiment 26, wherein at least some of the bristles extend to the internal surface of the conical member when in operative position.

[0311] 28. The apparatus of embodiments 26 or 27, wherein the impeller body comprises a tire brush.

[0312] 29. The apparatus of any one of embodiments 20-25, wherein the impeller body comprises one or more flanges extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position

[0313] 30. The apparatus of any one of embodiments 20-29, wherein the liquid is sterile water.

[0314] 31. The apparatus of any one of embodiments 20-30, further comprising a clamp to attach the conical member to the container such that the axis of the conical member is oblique to the rotational axis of the impeller.

[0315] 32. The apparatus of any one of embodiments 20-31, further comprising a stand configured to secure the rotary power source and the container in place during operation, the stand comprising: a base; a base securing member; an elongated member; and an upper member.

[0316] 33. The apparatus of any of embodiments 20-32, in combination with a second container is used for collection of the collected embryonic axes that have passed through the through-pores.

[0317] 34. The apparatus of any one of embodiments 20-33, wherein the container comprises: an internal volume of on the order of 10 L; a height on the order of 12 inches; and a diameter on the order of 10.5 inches.

[0318] 35. A system for automated separation and collection of embryonic axes of dicot seeds, the system comprising: means for rotating a suspension comprising a set of imbibed whole dicot seeds and a liquid relative to a surface having through-pores, the surface being effective to: comminute the set of imbibed whole dicot seeds into smaller pieces; and substantially pass embryonic axes; and means for collecting passed embryonic axes.

[0319] 36. The system of embodiment 35, wherein the surface comprises: an internal portion of an inverted three-dimensional conical member positioned in an internal volume of a container, the conical member having an internal space along an axis between an open top base end and aAgent Ref. No. P14771WOOO closed opposite bottom end; and wherein the internal portion is between the open top base end and the closed opposite bottom end.

[0320] 37. The system of embodiment 36, further comprising an impeller having: an interface operably connected to a controllable rotary power source; and an impeller body extending from a base end to an opposite end along a rotational axis, the impeller body configured to: rotate the suspension; and influence the set of imbibed whole dicot seeds toward or to the internal portion of the conical member.

[0321] 38. The system of embodiment 37, wherein the means for collecting comprises the container.

[0322] 39. The system of embodiment 38, wherein the means for collecting further comprises a second container, the second container configured to receive the passed embryonic axes from the first container.

[0323] 40. The system of any one of embodiments 37-39, further comprising a clamp to releasably attach the conical member to the container such that the axis of the conical member is oblique to the rotational axis of the impeller.

[0324] 41. The system of any one of embodiments 37-40, further comprising a stand configured to secure the rotary power source and the container in place during operation, the stand comprising: a base; a base securing member; an elongated member; and an upper member.

[0325] 42. The system of any one of embodiments 37-41, wherein the conical member is circular.

[0326] 43. The system of any one of embodiments 37-42, wherein the conical member is a truncated cone with the closed opposite bottom end closed at a truncation plane.

[0327] 44. The system of any one of embodiments 37-43, wherein the container comprises: an internal volume of on the order of 10 L; a height on the order of 12 inches; and a diameter on the order of 10.5 inches.

[0328] 45. The system of any one of embodiments 37-44, wherein the internal portion of the conical member and the impeller together are configured to degrade and / or comminute the set of imbibed whole dicot seeds.

[0329] 46. The system of any one of embodiments 37-45, wherein the impeller body has a cylindrical shape, conical shape, substantially cylindrical shape, or substantially conical shape.

[0330] 47. The apparatus of any one of embodiments 37-46, wherein the impeller body comprises one or more flanges extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.Agent Ref. No. P14771WOOO

[0331] 48. The system of any one of embodiments 37-46, wherein the impeller body comprises bristles extending generally laterally from the rotational axis towards the internal portion of the conical member when in operative position.

[0332] 49. The system of embodiment 48, wherein the bristles extend to the internal portion of the conical member when in operative position.

[0333] 50. The system of embodiments 48 or 49, wherein the impeller body comprises a tire brush.

[0334] 51. The system of any one of embodiments 35-50, wherein a pore size of each of the through-pores is on the order of or 6 millimeters.

[0335] 52. The system of any one of embodiments 35-51, wherein the liquid is sterile water.EXAMPLES

[0336] The following examples illustrate the effects of employing differing methods when performing embryonic axis extraction and collection as described herein.Example 1

[0337] Example 1 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 12-17, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 17 hours. The rotary power source was turned ON and spun at 330 RPM for 50 minutes. Testing used an Agrobacterium-based transformation system in sterile conditions. Embryonic axes obtained with the apparatus and system were compared to manually extracted embryonic axes. Using genotype TENG2284L and using 325 whole soybean seeds, 170 embryonic axes were collected. Eight uncracked seeds resulted. A transformation efficiency percentage (TE%) of 17.6% was obtained in embryonic axes obtained by automated extraction and a TE% of 22.4% was obtained by manual extraction.Example 2

[0338] Example 2 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 12-17, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 17.5 hours. The rotary power source was turned ON and spun at 315-350 RPM for 60 minutes. A new soybean genotype was tested. Using genotype TENE2324L and using 260 whole soybean seeds, 266 embryonic axes were collected. Eight uncracked seeds resulted. It is noted that issues withAgent Ref. No. P14771WOOO the seed counter required researchers to count seeds by hand, which likely caused the number of collected embryonic axes to be greater than that of the whole seeds. A transformation efficiency percentage (TE%) of 3.0% was obtained in embryonic axes obtained by automated extraction and a TE% of 15.5% was obtained by manual extraction.Example 3

[0339] Example 3 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 12-17, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed using a solid media and via water for 14 hours. The rotary power source was turned ON and spun at 300-325 RPM for 60 minutes. An Aldi grater (such as the conical member produced by Aldi described herein) with an unautoclaved brush was tested. A transformation efficiency percentage (TE%) of 4.5% was obtained in embryonic axes obtained by automated extraction, a TE% of 9.5% was obtained when using salt solution (GM045 68.57 g / L), and a TE% of 18.0% was obtained by manual extraction. GM045 is a salt (NaCl) solution used to aid in the filtration process. GM045 has a concentration of 68.57g / L. Using genotype NING1295 and using 325 whole soybean seeds, 134 embryonic axes were collected. The unautoclaved brush bruised explants and did not have as much regeneration as an autoclaved brush.Example 4

[0340] Example 4 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 12-17, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed using a solid media and via water for 14 hours. The rotary power source was turned ON and spun at 275-300 RPM for 50 minutes. An autoclaved brush (the brush used was the Drill Brush Power Scrubber by Useful Products 36D described herein, and the brush was autoclaved 1 time to soften the bristles), lower RPM, and one treatment of (GM046 34.285 g / L) were tested. A transformation efficiency percentage (TE%) of 0% was obtained in embryonic axes obtained by automated extraction, a TE% of 2.0% was obtained when using salt solution (GM046 34.285 g / L), and a TE% of 26.2% was obtained by manual extraction. Using genotype NING1295 and using 390 whole soybean seeds, 163 embryonic axes were collected. Twenty-five uncracked seeds resulted. Debris remained. Lots of bruising on both automation and salt treatment occurred. Some regeneration occurred.Agent Ref. No. P14771WOOOExample 5

[0341] Example 5 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 12-17, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed using a solid media and via water for 14 hours. The rotary power source was turned ON and spun at 300-325 RPM for 60 minutes. GM045 salt solution was tested after 20 minutes. Chlorine gas was applied to the brush and stopper. A plate was inadvertently dropped into the seeds, so the example may have been contaminated. A transformation efficiency percentage (TE%) of 16.5% was obtained in embryonic axes obtained by automated extraction, a TE% of 16.5% was obtained when using salt solution, and a TE% of 13.9% was obtained by manual extraction. Using genotype NINF1170 and using 390 whole soybean seeds, 200 embryonic axes were collected. The brush turned yellow due to the chlorine gas. Lots of bruising on both automation and more on salt treatment occurred. Lots of shoot regeneration on all treatments occurred.Example 6

[0342] Example 6 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 12-17, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed using a solid media and via water for 14 hours. The rotary power source was turned ON and spun at 260-290 RPM for 60 minutes. GM045 salt solution was tested after 20 minutes. Chlorine gas was applied to the brush and stopper. The brush was washed / rinsed off after a chemical reaction occurred with aluminum foil causing a liquid containing aluminum chloride to form on the brush. The washing / rinsing of the brush was an attempt to remove the liquid from the brush since aluminum chloride is known for its toxic properties, which could potentially affect the TE%. A relatively tall container was used, and a swirling motion was used when using the salt solution during the filtering process. A transformation efficiency percentage (TE%) of 4.3% was obtained in embryonic axes obtained by automated extraction, a TE% of 4.9% was obtained when using salt solution (GM045), and a TE% of 25.0% was obtained by manual extraction. Using genotype NINF1170 and using 390 whole soybean seeds, 281 embryonic axes were collected. Twenty- four uncracked seeds and 85 damaged embryonic axes resulted. As stated, a reaction occurred with aluminum foil. Three plates of contamination in salt treatment occurred. Some bruising on salt treatment occurred. Regeneration on all treatments occurred.Agent Ref. No. P14771WOOOExample 7

[0343] Example 7 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed using a solid media and via water for 14 hours. The rotary power source was turned ON and spun at 250-300 RPM for 35 minutes. A 3D-printed conical member (such as the conical member 1222) and table salt version of GM0045 was tested. A plate of seeds was dropped into the solution. A transformation efficiency percentage (TE%) of 0% was obtained in embryonic axes obtained by automated extraction, a TE% of 1.0% was obtained when using salt solution (GM045), and a TE% of 4.4% was obtained by manual extraction. Genotype GINE9252 and 520 whole soybean seeds were used. Two hundred twenty-one embryonic axes were collected. Foam formed on the liquid. Many embryonic axes were damaged. The seeds used were of a poor quality. The salt treatment turned yellow and had no regeneration. There was barely any regeneration on manual and automation treatments.Example 8

[0344] Example 8 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 17 hours. The rotary power source was turned ON and spun at 180-220 RPM for 30 minutes. GM047 salt solution was tested. Testing was conducted using a 3D-printed conical member (such as the conical member 1222) and a 10 liter container. Using genotype TENE2326L and using 650 whole soybean seeds, 494 embryonic axes were collected. Many embryonic axes were trapped in the conical member.Example 9

[0345] Example 9 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed using a solid media and via water for 14 hours. The rotary power source was turned ON and spun at 125-150 RPM for 45 minutes. A salt solution having a 5.15% concentration was tested. Testing was conducted using a 3D-printed conical member (such as the conical member 1222) and a 10 liter container. A transformation efficiency percentage (TE%) of 15.6% was obtainedAgent Ref. No. P14771WOOO in embryonic axes obtained by automated extraction, a TE% of 2.6% was obtained when using salt solution, and a TE% of 10.7% was obtained by manual extraction. Using genotype NING1295 and using 650 whole soybean seeds, 461 embryonic axes were collected.Example 10

[0346] Example 10 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 12-17, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed overnight (11 :00 am to 8:00 am) in Agrobacterium. An Aldi conical member was used for this example (such as the conical member produced by Aldi described herein). The rotary power source was turned ON and spun at 230-290 RPM for 40 minutes. A transformation efficiency percentage (TE%) of 0% was obtained in embryonic axes obtained by automated extraction. Using genotype NING1295 and using 390 whole soybean seeds, 281 embryonic axes were collected, 25 uncracked seeds resulted, and 84 damaged embryonic axes resulted. The embryonic axes experienced less damage and the seeds opened faster than usual.Example 11

[0347] Example 11 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed and put on a shaker for 22 hours. A twice-autoclaved brush sterilized using chlorine gas and a 3D-printed conical member (such as the conical member 1222), wherein the 3D-printed conical member was sterilized via autoclave, were tested. The rotary power source was turned ON and spun at 180-200 RPM for 40 minutes. A salt solution having a concentration of 34.3 g / L was tested. A transformation efficiency percentage (TE%) of 0% was obtained in embryonic axes obtained by automated extraction and a TE% of 3.4% was obtained when using salt solution. Using genotype NINF1170 and using 845 whole soybean seeds, 739 embryonic axes were collected.Example 12

[0348] Example 12 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed in solid media and via water for 14 hours. The seeds were not shaken. A thrice-autoclaved brushAgent Ref. No. P14771WOOO and a 3D-printed conical member (such as the conical member 1222) were tested. The rotary power source was turned ON and spun at 175-200 RPM for 45 minutes. A salt solution having a concentration of 34.285 g / L was tested. A transformation efficiency percentage (TE%) of 6.7% was obtained in embryonic axes obtained by automated extraction, a TE% of 0.3% was obtained when using salt solution, and a TE% of 20% was obtained by manual extraction. Using genotype NING1295 and using 650 whole soybean seeds, 410 embryonic axes were collected.Example 13

[0349] Example 13 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed in solid media and via water for 14 hours. The seeds were not shaken. A thrice-autoclaved brush and a 3D-printed conical member (such as the conical member 1222) were tested. The rotary power source was turned ON and spun at 160-200 RPM for 45 minutes. A salt solution having a concentration of 25.7 g / L was tested. A transformation efficiency percentage (TE%) of 5.9% was obtained in embryonic axes obtained by automated extraction, a TE% of 2.0% was obtained when using salt solution, and a TE% of 8.9% was obtained by manual extraction. Using genotype NING1295 and using 650 whole soybean seeds, 438 embryonic axes were collected, 11 uncracked seeds resulted, and 201 damaged embryonic axes resulted.Example 14

[0350] Example 14 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed on a shaker for 21 hours. A thrice-autoclaved tapered brush and a 3D-printed conical member (such as the conical member 1222) were tested. The rotary power source was turned ON and spun at 170-200 RPM for 45 minutes. A salt solution having a concentration of 17.1 g / L was tested. Using genotype NINF1170 and using 650 whole soybean seeds, 536 embryonic axes were collected, 5 uncracked seeds resulted, and 109 damaged embryonic axes resulted. Example 14 resulted in Agrobacterium overgrowth on explants. Overall, salt explants are not healthy.Example 15

[0351] Example 15 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well asAgent Ref. No. P14771WOOOParagraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 19.5 hours. A tapered brush autoclaved 4 times and a 3D-printed conical member (such as the conical member 1222) were tested. Mannitol and sorbitol were compared during filtration at 10.00 g / L. The rotary power source was turned ON and spun at 175-225 RPM for 80 minutes. Using genotype NING1295 and using 650 whole soybean seeds, 297 embryonic axes were collected. After autoclaving the brush 4 times, the brush struggled to break open the seed coat and the brush is not as effective as other Examples. Mannitol and sorbitol caused swelling of embryonic axes moving from co-culture (CC) to SIMl. Contamination resulted for all treatments. Orange fungus appeared on manual, and white fungus appeared on mannitol.Example 16

[0352] Example 16 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed on solid media and via water for 14 hours. A paint mixer with 50% glycerol and 50% water was tested. The rotary power source was turned ON and spun at 175 RPM for 10 minutes. Using genotype GINF9543 and using 650 whole soybean seeds, 306 embryonic axes were collected. The glycerol treatment started to form callus and did not regenerate.Example 17

[0353] Example 17 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 20.5 hours. A paint mixer was tested with the 60L Overhead Stirrer produced by +UXI 34B, described herein, at lower RPMs. No filtration chemical was used. The conical member was oscillated up and down at the 20 minute and 40 minute marks using 2 forceps. The rotary power source was turned ON and spun at 150 RPM for 40 minutes. Using genotype GINF9812 and using 585 whole soybean seeds, 349 embryonic axes were collected, 50 uncracked seeds resulted, and 186 damaged embryonic axes resulted. The glycerol treatment started to form callus and did not regenerate. Seed quality was poor. At least 45 seeds were trapped at the bottom of the conical member. When plating on co-culture (CC), the automatically, or at least semi-automatically, extracted embryonic axes were longer and straightened out. After one week, the automatically, or at least semi-automatically, extracted embryonic axes looked like the manually extracted embryonic axes.Agent Ref. No. P14771WOOOExample 18

[0354] Example 18 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 21 hours. A twice-autoclaved brush was tested with the 60L Overhead Stirrer produced by +UXI 34B, described herein, at lower RPMs. No filtration chemical was used. The conical member was oscillated up and down at the 20 minute and 40 minute marks using 2 forceps. The rotary power source was turned ON and spun at 150 RPM for 45 minutes. Using genotype GIND9185 and using 821 whole soybean seeds, 486 embryonic axes were collected, 33 uncracked seeds resulted, and 302 damaged embryonic axes resulted. There was possible contamination due to water flowing over chuck. Seed quality was poor. Manually cut embryonic axes were not imbibed in enough water and needed extra time to imbibe.Example 19

[0355] Example 19 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 21 hours. The Paint Mixer Drill Attachment produced by Edward Tools 36B was tested with the 60L Overhead Stirrer produced by +UXI 34B, both described herein, at lower RPMs. The rotary power source was turned ON and spun at 125 RPM for 45 minutes. Using genotype GING9244 and using 577 whole soybean seeds, 411 embryonic axes were collected and 13 uncracked seeds resulted.Example 20

[0356] Example 20 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

[0214] -

[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 21 hours. The Paint Mixer Drill Attachment produced by Edward Tools 36B was tested with the 60L Overhead Stirrer produced by +UXI 34B, both described herein, at lower RPMs. A 5 liter container was used. Glycerin was tested with water and liquid co-culture (CC). The rotary power source was turned ON and spun at 125-150 RPM for 45 minutes. Using genotype NING1295 and using 780 whole soybean seeds, 242 embryonic axes were collected and 50 uncracked seeds resulted. A small amount of water was left during the time between extractionAgent Ref. No. P14771WOOO and infection. The extraction process was not as efficient with the 5 liter beaker as compared to other Examples. Inefficiency may have been due to small seed size clogging the bottom row of through-pores of the conical member.Example 21

[0357] Example 21 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30 as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). Whole soybean seeds were imbibed for 19 hours. The rotary power source was turned ON and spun at 100 RPM for 50 minutes. Using genotype NING1295 and using 650 whole soybean seeds, 433 embryonic axes were collected and 7 uncracked seeds resulted. Some of the bottom through-pores of the conical member were clogged by cotyledons. Quite a few embryonic axes were left in the conical member.Example 22

[0358] Example 22 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-G, 26, and / or 30, as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). The impeller used was the Paint Mixer Drill Attachment produced by Edward Tools 36B. Using 800 whole soybean seeds, 689 embryonic axes were extracted. The extraction time was 40 to 60 minutes. The speed of the impeller during extraction was 75 RPM. Manual mixing / stirring was required, wherein said manual mixing / stirring included 3 repetitions every 20 minutes during extraction.Example 23

[0359] Example 23 illustrates the effects of performing automated embryonic extraction using an apparatus and / or system described herein per the following specifications. The apparatus used and workflow were essentially as shown in Figures 19A-26, and / or 30, as well as Paragraphs

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[0258] (methods 310, 410, and / or 510). The impeller used was the impeller 1336 shown in at least Figure 21. Using 800 whole soybean seeds, 672 embryonic axes were extracted. The extraction time was 25 to 30 minutes. The speed of the impeller during extraction was 60 to 75 RPM. Manual mixing / stirring was not required.

[0360] From the foregoing, it can be seen that the present disclosure accomplishes at least all of the stated objectives.

Claims

Agent Ref. No. P14771WOOOCLAIMSWhat is claimed is:

1. A method for automated separation and collection of embryonic axes of dicot seeds, the method comprising: a) rotating a suspension of a plurality of whole dicot seeds and a liquid relative to a surface having through-pores, the surface being effective to: i) comminute the whole dicot seeds into smaller pieces, and ii) substantially pass embryonic axes; and b) collecting the passed embryonic axes.2 The method of claim 1, wherein the plurality of whole dicot seeds comprises a plurality of seeds of the Fabaceae family.3 The method of claim 1, wherein the comminution imposes on the whole dicot seeds one or more of: a) friction; b) shear or impact stress(es); and / or c) degradation.4 The method of claim 1, further comprising, while rotating the suspension, influencing the seeds toward or to the surface.5 The method of claim 4, wherein the influencing comprises one or more of: a) centrifugal force(s); b) mechanical force(s); and / or c) gravitational force(s).6 The method of claim 1, wherein the surface comprises: an internal portion of an inverted three-dimensional conical member positioned in an internal volume of a container, the conical member having an internal space along an axis between an open top base end and a closed opposite bottom end; and wherein the internal portion is between the open top base end and the closed opposite bottom end.Agent Ref. No. P14771WOOO7. The method of claim 6, wherein the rotating comprises rotating the suspension with an impeller: a) operably connected to a rotary power source; and b) wherein the impeller comprises an impeller body having top and bottom ends along a rotational axis, wherein the impeller body is adjustable relative to the closed opposite bottom end of the conical member.8 The method of claim 7, wherein the impeller body comprises: a) a structure extending towards or to the internal portion of the conical member when in operative position; b) bristles extending generally laterally from the rotational axis towards, at, or near the internal portion of the conical member when in operative position; or c) one or more flanges extending generally laterally from the rotational axis towards, at, or near the internal portion of the conical member when in operative position.9 The method of claim 8, further comprising: a) autoclaving the impeller body prior to the rotation of the suspension; and b) sterilizing the impeller body prior to the rotation of the suspension.10 The method of claim 8, further comprising adjusting height of the bottom end of the impeller body such that the bottom end of the impeller body is positioned within several inches from the closed opposite bottom end of the conical member.11 The method of claim 6, further comprising securing the conical member to the container via a clamp prior to the rotating of the suspension.12 The method of claim 6, wherein the collecting of the passed embryonic axes further comprises at least one of the following steps: a) removing the conical member from the internal volume of the container and pouring out excess liquid from the container wherein at least a portion of debris is poured out with the excess liquid but the passed embryonic axes generally remain in the container; b) adding an additional amount of liquid to the container;Agent Ref. No. P14771WOOO c) moving a suspension remaining in the container wherein said remaining suspension comprises at least a portion of the liquid, the additional amount of liquid, the passed embryonic axes, and another portion of the debris; d) pouring the remaining suspension into a second container; e) pouring out a majority of liquid from the second container; f) moving the second container to separate the passed embryonic axes in the second container from debris in the second container; g) pouring out the debris from the second container while the passed embryonic axes remain in the second container; h) resuspending the passed embryonic axes in the second container; and / or i) pouring the passed embryonic axes into a third container.

13. The method of claim 12, wherein the additional amount of liquid is 500 milliliters.

14. The method of claim 12, wherein the third container is a petri dish.

15. The method of claim 12, further comprising suspending the passed embryonic axes in a tissue culture medium.

16. The method of claim 1, wherein: a) the plurality of whole dicot seeds are soybean seeds; b) a pore size of each of the through-pores is on the order of or 6 millimeters; and c) the suspension is rotated on the order of 75 to 200 or 300 RPM.

17. The method of claim 1, wherein the collecting of the passed embryonic axes is for the purpose of one or more of: a) a biolistic transformation; b) biolistic gene delivery; c) an agrobacterial transformation; and / or d) gene editing.

18. The method of claim 1, wherein the whole dicot seeds are imbibed for several hours prior to being rotated in the suspension.Agent Ref. No. P14771WOOO19. The method of claim 1, wherein the liquid is sterile water.

20. An apparatus for automated separation and collection of embryonic axes of dicot seeds, the apparatus comprising: a) a container having: i) an internal volume configured to receive and retain a suspension of a set of imbibed whole dicot seeds and a liquid; b) an inverted three-dimensional conical member positioned at least partially in the internal volume of the container, the conical member having: i) an internal space along an axis between an open top base end and a closed opposite bottom end; and ii) an internal surface between the open top base end and the closed opposite bottom end with through-pores, the internal surface being effective to:1) comminute the imbibed whole dicot seeds; and2) substantially pass embryonic axes of each of the imbibed whole dicot seeds; c) an impeller having: i) an interface configured to operably connect to a rotary power source; and ii) an impeller body extending from a base end to an opposite end along a rotational axis, the impeller body configured to:1) rotate the suspension; and2) influence the set of imbibed whole dicot seeds toward or to the internal surface of the conical member; and d) a collection area in the container outside the internal space of the conical member to collect embryonic axes that have passed through the through-pores.

21. The apparatus of claim 20, wherein the conical member is a circular conical member.

22. The apparatus of claim 20, wherein the conical member is a truncated cone with the closed opposite bottom end closed at a truncation plane.

23. The apparatus of claim 20, wherein the internal surface and the impeller together are configured to degrade and / or comminute the set of imbibed whole dicot seeds.Agent Ref. No. P14771WOOO24. The apparatus of claim 20, wherein a pore size of each of the through-pores is on the order of or 6 millimeters.

25. The apparatus of claim 20, wherein the impeller body has a cylindrical shape, conical shape, substantially cylindrical shape, or substantially conical shape.

26. The apparatus of claim 20, wherein the impeller body comprises bristles extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.

27. The apparatus of claim 26, wherein at least some of the bristles extend to the internal surface of the conical member when in operative position.

28. The apparatus of claim 26, wherein the impeller body comprises a tire brush.

29. The apparatus of claim 20, wherein the impeller body comprises one or more flanges extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.

30. The apparatus of claim 20, wherein the liquid is sterile water.

31. The apparatus of claim 20, further comprising a clamp to attach the conical member to the container such that the axis of the conical member is oblique to the rotational axis of the impeller.

32. The apparatus of claim 20, further comprising a stand configured to secure the rotary power source and the container in place during operation, the stand comprising: a) a base; b) a base securing member; c) an elongated member; and d) an upper member.

33. The apparatus of claim 20, in combination with a second container is used for collection of the collected embryonic axes that have passed through the through-pores.Agent Ref. No. P14771WOOO34. The apparatus of claim 20, wherein the container comprises: a) an internal volume of on the order of 10 L; b) a height on the order of 12 inches; and c) a diameter on the order of 10.5 inches.

35. A system for automated separation and collection of embryonic axes of dicot seeds, the system comprising: a) means for rotating a suspension comprising a set of imbibed whole dicot seeds and a liquid relative to a surface having through-pores, the surface being effective to: i) comminute the set of imbibed whole dicot seeds into smaller pieces; and ii) substantially pass embryonic axes; and b) means for collecting passed embryonic axes.

36. The system of claim 35, wherein the surface comprises: an internal portion of an inverted three-dimensional conical member positioned in an internal volume of a container, the conical member having an internal space along an axis between an open top base end and a closed opposite bottom end; and wherein the internal portion is between the open top base end and the closed opposite bottom end.

37. The system of claim 36, further comprising an impeller having: a) an interface operably connected to a controllable rotary power source; and b) an impeller body extending from a base end to an opposite end along a rotational axis, the impeller body configured to: i) rotate the suspension; and ii) influence the set of imbibed whole dicot seeds toward or to the internal portion of the conical member.

38. The system of claim 37, wherein the means for collecting comprises the container.

39. The system of claim 38, wherein the means for collecting further comprises a second container, the second container configured to receive the passed embryonic axes from the first container.Agent Ref. No. P14771WOOO40. The system of claim 37, further comprising a clamp to releasably attach the conical member to the container such that the axis of the conical member is oblique to the rotational axis of the impeller.

41. The system of claim 37, further comprising a stand configured to secure the rotary power source and the container in place during operation, the stand comprising: a) a base; b) a base securing member; c) an elongated member; and d) an upper member.

42. The system of claim 37, wherein the conical member is circular.

43. The system of claim 37, wherein the conical member is a truncated cone with the closed opposite bottom end closed at a truncation plane.

44. The system of claim 37, wherein the container comprises: a) an internal volume of on the order of 10 L; b) a height on the order of 12 inches; and c) a diameter on the order of 10.5 inches.

45. The system of claim 37, wherein the internal portion of the conical member and the impeller together are configured to degrade and / or comminute the set of imbibed whole dicot seeds.

46. The system of claim 37, wherein the impeller body has a cylindrical shape, conical shape, substantially cylindrical shape, or substantially conical shape.

47. The apparatus of claim 37, wherein the impeller body comprises one or more flanges extending generally laterally from the rotational axis towards the internal surface of the conical member when in operative position.Agent Ref. No. P14771WOOO48. The system of claim 37, wherein the impeller body comprises bristles extending generally laterally from the rotational axis towards the internal portion of the conical member when in operative position.

49. The system of claim 48, wherein the bristles extend to the internal portion of the conical member when in operative position.

50. The system of claim 48, wherein the impeller body comprises a tire brush.

51. The system of claim 35, wherein a pore size of each of the through-pores is on the order of or 6 millimeters.

52. The system of claim 35, wherein the liquid is sterile water.

53. System(s), method(s), apparatus(es), or kit(s) as herein described.

54. System(s), method(s), apparatus(es), or kit(s) incorporating novel and unobvious aspect(s) of the present disclosure as they are as substantially shown or described.

55. Alternative system(s), method(s), apparatus(es), or kit(s) incorporating other novel and unobvious aspect(s) of the present disclosure as they are as substantially shown or described.

56. The system(s), method(s), apparatus(es), or kit(s) according to any one of the preceding claims further comprising improvement(s), structure(s), and / or step(s) known to one of ordinary skill in the art.

57. The system(s), method(s), apparatus(es), or kit(s) according to any one of the preceding claims wherein an element of the same possesses a functional capability as substantially shown or described and / or a functional capability known to one of ordinary skill in the art.

58. The system(s), method(s), apparatus(es), or kit(s) according to any one of the preceding claims within its applicable or intended environment.

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