Material transfer device and method of using the same

Abstract

Transfer systems, devices, kits and methods described herein can be used to facilitate the transfer of materials (e.g., fluids) to or from supply containers, such as being transferred to or from the holes of a supply plate, while keeping the target agent attached to the holes. The system can include a supply plate, a transfer adapter and a receiving plate, which can be coupled to form a transfer assembly. The transfer adapter can include a flat sheet and a plurality of openings, and it can be configured to regulate the flow of the fluid flowing out of the hole. The transfer assembly can be placed in a centrifuge, and the fluid transfer force generated by the centrifuge can cause the fluid to flow out of the hole of the supply plate, through the opening in the transfer adapter, and into the receiving plate.

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Provisional Patent Application Serial No. 62 / 911,811, filed on October 7, 2019, the entire contents of which are incorporated herein by reference for all purposes. Technical Field

[0003] The present invention generally relates to tools for material transfer, such as transfer of fluids into and / or out of a porous plate, for example decanting and / or loading a porous plate, or transferring liquids between porous plates or between a porous plate and another structure. Background Art

[0004] Porous plates are used for a wide range of laboratory applications, such as multiple samples are tested in parallel for chemistry, biology or pharmacology. Porous plates may have a grid of small open holes or holes, and each hole can accommodate different samples for evaluation. For some evaluations, the sample may include a target agent fixed on the base of each hole. These target agents may include but are not limited to proteins, nucleic acids, cells, microorganisms (e.g., bacteria, fungi), plants (e.g., algae), viruses, small molecule drugs or other chemical compounds, or antigen-antibody complexes. In general, the sample in the evaluation also includes a fluid that partially fills each hole and immerses the target agent. After reacting, combining or other processes in the hole, it is usually desired to test the fluid composition and / or target agent of each sample respectively, or replace or supplement the fluid composition of each sample with a new fluid. Therefore, a technology has been developed to transfer fluid to the hole of a porous plate or to transfer from the hole of a porous plate while keeping at least some target agents attached to the hole.

[0005] Although several techniques and devices have been developed to facilitate the transfer of fluids to or from the holes of a porous plate, they all have significant disadvantages. For example, vacuum suction is commonly used to remove fluids in both manual and automatic / robotic techniques. However, the force and flow generated by a suction device (such as a pipette) are unevenly distributed in the hole; the suction and flow are greatest near the tip of the suction device. Therefore, vacuum suction can affect different parts of the sample differently, which can introduce undesirable variability within a sample and between different samples. For example, a target agent near the tip of the suction device may be accidentally damaged or separated and removed from the hole, while a target agent away from the tip may not be affected.

[0006] In addition, manual techniques are usually slow and laborious, even if a multichannel device is used to load or remove fluid from more than one hole at a time. This is especially true if the fluid is transferred to or from a multiwell plate with hundreds of holes. Manual devices generally do not have enough channels to transfer fluid to or from all holes at the same time, and some processes occurring in the holes may be time-sensitive. Therefore, transferring fluid to different holes or transferring from different holes at different times may be another source of undesirable variability between different samples. In addition, the use of a multichannel device for fluid transfer can cause cross contamination between the holes of a multiwell plate. Automation or robotic devices such as liquid handlers generally have the disadvantage of being more expensive.

[0007] Therefore, there is the remarkable demand of the improved device or system that is convenient for fluid to be transferred to the hole of porous plate or to be transferred from the hole of porous plate.This device or system should be able to transfer fluid in each hole or to be transferred out from each hole simultaneously, keep target agent to be fixed in the hole and be undisturbed basically simultaneously.The force involved in the fluid transfer should be consistent in each hole and between different holes, to reduce the variability between fluid sample or target agent.In addition, improved fluid transfer device or system should be relatively low in cost, but need minimum manual work to realize the result of expectation. Summary of the invention

[0008] In one aspect, a device for transferring a material such as a fluid is disclosed herein, comprising: a planar sheet having a first planar surface, a second planar surface, and a plurality of openings, wherein each of the plurality of openings extends between the first planar surface and the second planar surface; a plurality of main extensions, wherein each of the plurality of main extensions protrudes from the first planar surface and comprises a main lumen; and a plurality of secondary extensions, wherein each of the plurality of secondary extensions protrudes from the second planar surface and comprises a secondary lumen, and wherein each secondary lumen is aligned with the opening and the main lumen to produce a continuous transfer lumen. In some embodiments, each of the plurality of main extensions in the device has an inner cross-sectional area and an outer cross-sectional area, and the inner cross-sectional area is greater than the outer cross-sectional area. In any of the foregoing embodiments, each main extension may have at least two regions, and each of the at least two regions may have a different angle relative to the planar sheet. In some embodiments, the at least two regions include a first region close to the planar sheet and a second region away from the planar sheet, and the angle of the first region relative to the planar sheet is greater than the angle of the second region relative to the planar sheet.

[0009] In any of the foregoing embodiments, each of the plurality of main extensions may include at least one structure configured to fluidically connect to the main lumen. In some embodiments, the at least one structure includes an orifice, a hole, a slit, a gap, a notch, a groove, or a channel. In some embodiments, the main extension includes four slits configured to fluidically connect to the main lumen.

[0010] In one aspect, a system for transferring a fluid is disclosed herein, comprising: a transfer adapter, wherein the transfer adapter comprises a first side, a second side, and a plurality of openings, and wherein each of the plurality of openings extends between the first side and the second side; and a receiving plate, wherein the receiving plate is configured to be removably connected to the second side of the transfer adapter. In some embodiments, the system further comprises a supply plate, wherein the supply plate comprises a hole or a plurality of holes, and wherein the supply plate is configured to be removably coupled to the first side of the transfer adapter. In some embodiments, when the transfer adapter and the supply plate are removably coupled, each of the plurality of openings is aligned with a different hole in the plurality of holes.

[0011] In any of the foregoing embodiments, the transfer adapter may include a plurality of main extensions. In any of the foregoing embodiments, each of the plurality of main extensions may be configured to seal against an inner surface of a different one of the plurality of holes of the feed plate.

[0012] In any of the foregoing embodiments, the receiving plate may include a plurality of receiving holes. In any of the foregoing embodiments, when the transfer adapter and the receiving plate are removably coupled, each of the plurality of openings may be aligned with a different one of the plurality of receiving holes.

[0013] In any of the foregoing embodiments, the transfer adapter may include a plurality of secondary extensions, and each of the plurality of secondary extensions may be configured to be inserted into a different one of the plurality of receiving holes.

[0014] In any of the foregoing embodiments, the transfer adapter may include an adhesive on the first side and / or the second side.

[0015] In any of the foregoing embodiments, the transfer adapter can include a plurality of leaflets adjacent each of the plurality of openings, and at least one leaflet can be movable between an open position and a closed position.

[0016] In any of the foregoing embodiments, the transfer adapter may be configured to allow fluid to flow through the plurality of openings only when an external force is applied to the fluid.

[0017] In one aspect, disclosed herein is a method for transferring a material such as a fluid, comprising: coupling a transfer adapter, a supply plate, and a receiving plate to form a transfer assembly in a first position (e.g., an upright position), wherein in the first position, the transfer adapter is positioned below the receiving plate and above the supply plate; and centrifuging the transfer assembly about a rotational axis, wherein when the transfer assembly is centrifuged, the supply plate is positioned closer to the rotational axis than the transfer adapter and the receiving plate, wherein the transfer adapter includes a plurality of openings and the supply plate includes a plurality of holes, and wherein each of the plurality of openings is aligned with a different hole of the plurality of holes when the transfer assembly is in the first position and when the transfer assembly is centrifuged. In some embodiments, the method further comprises separating the supply plate from the transfer adapter.

[0018] In any of the foregoing embodiments, when the supply plate and the transfer adapter are coupled, a seal can be formed between each hole in the plurality of holes and the transfer adapter. In some embodiments, the seal allows fluid to flow from the plurality of holes through the plurality of openings, but prevents fluid from flowing between holes in the plurality of holes.

[0019] In any of the foregoing embodiments, each well in the plurality of wells can include a target agent attached to the well, and a fluid. In some embodiments, the target agent remains attached to the well after centrifugation.

[0020] In any of the foregoing embodiments, the fluid may be transferred to a receiving plate after centrifugation.

[0021] In any of the foregoing embodiments, the receiving plate may include a plurality of receiving holes, and each of the plurality of openings may be aligned with a different one of the plurality of receiving holes when the transfer assembly is in the first position and when the transfer assembly is centrifuged. In some embodiments, when the receiving plate and the transfer adapter are coupled, a seal is formed between each of the plurality of receiving holes and the transfer adapter. In some embodiments, the seal allows fluid to flow through the plurality of openings to the plurality of receiving holes, but prevents fluid from flowing between the plurality of receiving holes.

[0022] In one aspect, disclosed herein is a method for transferring a material, such as a fluid, from a well of a supply plate comprising a plurality of wells and / or transferring a material, such as a fluid, to a well of a supply plate comprising a plurality of wells, wherein at least one well comprises a plurality of target agents attached to the well, and a fluid, comprising: applying a fluid transfer force to the supply plate, wherein the fluid transfer force has a simultaneous and substantially uniform effect on the plurality of target agents in the at least one well. In some embodiments, each of at least two of the plurality of wells comprises a plurality of target agents attached to the well, and a fluid, and wherein the fluid transfer force has a simultaneous and substantially uniform effect on the plurality of target agents in each of the at least two wells.

[0023] In one aspect, disclosed herein is a device for transferring a material such as a fluid, comprising: a planar sheet having a first planar surface on a first side and a second planar surface on a second side, a plurality of compartments (e.g., holes) on the first side having an opening on the second side, a plurality of extensions on the second side protruding from the second planar surface and including lumens connected to the openings of the compartments. In some embodiments, each extension is configured to be inserted into a different receiving hole of a receiving plate. In some embodiments, each extension is configured to seal against a different receiving hole of a receiving plate.

[0024] In any of the foregoing embodiments, an inner surface of the extension may be configured to form an angle with an inner wall of the receiving hole, and the angle may be approximately 7 degrees or less.

[0025] In any of the foregoing embodiments, the device may further comprise reagents, such as liquid reagents, eg, lyophilized reagents, in the plurality of compartments.

[0026] In any of the foregoing embodiments, each compartment of the device can include a structure configured to hold and / or dispense a reagent. In some embodiments, the structure includes a protrusion.

[0027] On the one hand, disclosed herein is a method for transferring reagents, comprising: connecting the device of any one of the aforementioned embodiments to a receiving plate to form a transfer assembly in a first position (e.g., an upright position); and centrifuging the transfer assembly around a rotation axis, wherein when the transfer assembly is centrifuged, the device is positioned closer to the rotation axis than the receiving plate, wherein each opening of the device is aligned with a different receiving hole of the receiving plate, thereby transferring the reagent in at least one compartment of the device to the corresponding receiving hole.

[0028] In any of the foregoing embodiments, the transfer systems, devices, kits, and methods described herein can be used to facilitate the transfer of fluids to or from the holes of a porous plate while keeping the target agent attached to the hole. On the one hand, a transfer adapter for transferring materials such as fluids is disclosed herein. In a particular embodiment, the system may include a porous plate and a transfer adapter, which are separated from each other, or connected to form a transfer assembly. On the one hand, the porous plate and the transfer adapter are reversibly connected. On the one hand, the porous plate and the transfer adapter are sealingly connected to prevent leakage between the porous plate and the transfer adapter. In any of the foregoing embodiments, the system may further include a receiving plate. The receiving plate can be separated from the transfer adapter, or can be connected to the transfer adapter and / or the porous plate to form a transfer assembly. On the one hand, the transfer adapter and the receiving plate are reversibly connected. On the one hand, the transfer adapter and the receiving plate are sealingly connected to prevent leakage between the transfer adapter and the receiving plate.

[0029] In certain embodiments, the porous plate can be a standard off-the-shelf porous plate, or as described in more detail below, it can be customized in one or more ways. The transfer adapter can include a flat sheet and a plurality of openings, and it can be configured to regulate the flow of the fluid out of the hole. The receiving plate can include one or more receiving holes, and the transferred fluid can be accommodated in the one or more receiving holes. The transfer assembly can be placed in a centrifuge, and the fluid transfer force generated by the centrifuge can cause the fluid to flow out of the hole of the porous plate, through the opening in the transfer adapter, and into the one or more receiving holes of the receiving plate. The fluid transfer force can be applied consistently in each hole and between different holes, and the fluid can be transferred from each hole at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1A and 1C is a perspective view of an embodiment of a transfer system described herein. Figure 1B and 1D is a perspective view of an embodiment of a transfer assembly described herein.

[0031] Figure 2 is an exemplary depiction of an embodiment of wells of a multi-well plate described herein.

[0032] Figure 3A and 3B is a perspective view of an embodiment of a supply plate described herein. Figure 3A An embodiment of a separation hole structure inserted into a holding cavity of a feed plate is shown, and Figure 3B The feed plate is shown with the separation hole structure removed.

[0033] Figures 4A-4D is a perspective view of an embodiment of a separation pore structure described herein. Figure 4D is a close-up perspective view of one embodiment of a separation hole structure.

[0034] Figure 5A and 5B is a perspective view of an embodiment of a guide hole structure described herein. Figure 5C is a cross-sectional view of the guide hole structure described in this article.

[0035] Fig. 6A and 6B is a perspective view of an embodiment of a transfer adapter described herein. Figure 6C is a close up perspective view of a first side of the transfer adapter. Fig.6D is a cross-sectional view of a first side of a transfer adapter in an embodiment inserted into a well of a multi-well plate. Fig. 6E is a close up perspective view of the second side of the transfer adapter. Fig. 6F and 6G is a cross-sectional view of a second side of a transfer adapter in an embodiment inserted into a receiving hole of a receiving plate.

[0036] Fig. 7A is a perspective view of an embodiment of a transfer adapter and a multiwell plate described herein. Figure 7B A close-up perspective view of a portion of a transfer adapter. Figure 7C An example of using centrifugal force to transfer fluid into a receiving hole is shown. Fig.7D It is shown that the multiple subdivided regions disclosed herein help reduce interference with unequal meniscus structures.

[0037] Figures 8A-8C are side views of embodiments of transfer assemblies described herein in different orientations.

[0038] Fig. 9 Systems and methods for material transfer described herein are shown.

[0039] Fig. 10A and 10B Systems and methods for material transfer described herein are shown. DETAILED DESCRIPTION

[0040] In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments. However, those skilled in the art will appreciate that the compounds of the present invention can be made and used without these details. In other cases, well-known structures are not shown or described in detail to avoid unnecessarily obscuring the description of the embodiments. Unless the context otherwise requires, in the subsequent description and claims, the word "comprise" and its variants, such as "comprises" and "comprising", should be interpreted in an open, inclusive sense, that is, "including but not limited to". In addition, the term "comprising" (and related terms, such as "comprise" or "comprises" or "having" or "including") is not intended to exclude certain other embodiments, for example, any embodiment of the composition of matter, composition, method or process, etc. described herein can be "composed" or "essentially composed of" the features described. The headings provided herein are for convenience only and do not explain the scope or meaning of the claimed embodiments.

[0041] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout the specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0042] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a hole" includes one or more holes. Furthermore, the terms "or" as used herein are understood to be inclusive and cover "or" and "and" unless otherwise specified or obvious from the context.

[0043] In some embodiments, the present disclosure relates to leak-proof or fluid-closed systems, particularly when assembling a transfer system for transfer, such as using a centrifuge. A "leak-proof" or "fluid-closed" system refers to a system comprising one or more containers (e.g., wells), chambers, valves, and / or passages (which may be interconnected and in communication with each other) in which liquid within the system cannot communicate with the outside of such system, and likewise, liquid outside such system cannot communicate with liquid contained within the system.

[0044] Transfer system, device, kit and method as described herein can be used for facilitating fluid transfer to the hole of porous plate or transfer from the hole of porous plate.Described system can comprise supply plate, transfer adapter and receiving plate, and they can be connected to form transfer assembly.In certain embodiments, supply plate and / or receiving plate can be the porous plate of any standard that can be used for various laboratory applications, and its embodiment is well-known in the art.However, in other embodiments, supply plate and / or receiving plate can comprise one or more features so as to facilitate the interaction with other members of transfer system or allow the division or combination of hole.Supply plate and / or receiving plate can comprise any number of holes, including but not limited to standard 6 holes, 24 holes, 96 holes and 384 holes arrangement.At least some holes can be filled with fluid and have one or more target agents attached to its inner surface, such as but not limited to protein, nucleic acid, cell, microorganism (for example, bacterium, fungus), plant (for example, algae), virus, small molecule drug or any compound, antigen-antibody complex etc.

[0045] In order to transfer fluid to or from the hole in a controlled manner, the supply plate and / or the receiving plate can be releasably connected to the transfer adapter. The transfer adapter may include a flat sheet and a plurality of openings, and the plurality of openings are, for example, arranged so that each opening is aligned with different holes of the supply plate and / or the receiving plate. The transfer adapter can be sealed against the supply plate and / or the receiving plate in a way so that the fluid leaving the supply plate hole can flow out through the opening, but will not flow into another hole of the supply plate, and / or the fluid entering the receiving plate hole can flow in through the opening, but will not flow into another hole of the receiving plate. One or more features of the transfer adapter can facilitate the alignment of the opening with the hole and / or the sealing of the plate. For example, each opening may have an associated extension that protrudes from the transfer adapter surface and fits into the hole of the supply plate. The transfer adapter may also include one or more features to facilitate alignment with the receiving plate and / or sealing against the receiving plate.

[0046] The fluid transferred from the hole of the supply plate can flow through the opening in the transfer adapter and enter the receiving plate. In some embodiments, the receiving plate can allow the fluid transferred from each hole to be mixed, for example, by containing all fluids in a fluidly connected area (e.g., a single large hole). In other embodiments, the receiving plate can be configured to prevent the fluids transferred from the supply plate from mixing, and the fluids transferred from each hole can be contained in a separate area of ​​the receiving plate. For example, the receiving plate can include a plurality of receiving holes, and the receiving holes can be aligned with the opening of the transfer adapter and sealed against the opening of the transfer adapter. If the transferred fluid is to be studied, this may be particularly useful.

[0047] The method described herein may include forming a transfer assembly and a centrifugal transfer assembly. The transfer assembly may be formed by directional coupling of a supply plate, a transfer adapter, and a receiving plate with a transfer adapter positioned above the supply plate and below the receiving plate. This coupling may involve aligning the opening of the transfer adapter with the hole of the supply plate and sealing the supply plate and the transfer adapter together. Similarly, in an embodiment of a receiving plate including a plurality of receiving holes, coupling the receiving plate and the transfer adapter may include aligning the opening and the receiving hole and sealing the receiving plate and the transfer adapter together. Then, the transfer assembly may be repositioned and centrifuged around the axis of rotation. When the centrifugal transfer assembly is centrifuged, it may be oriented so that the supply plate is closer to the axis of rotation than the transfer adapter and the receiving plate. The transfer assembly may be centrifuged at a desired speed and for a desired duration to transfer substantially all of the fluid from the hole of the supply plate to the receiving plate while keeping the target agent attached to the hole. Then, the plate of the transfer assembly may be separated, and the isolated target agent and / or the fluid of the transfer may be taken.

[0048] Throughout this disclosure, for exemplary purposes, reference is made to fluid transfer or liquid transfer. It should be understood that the present transfer system and method can be used to transfer one or more reagents in any suitable form, such as, but not limited to, liquids, solutions, gels (e.g., polymeric gels), powders (e.g., dry powders such as lyophilized powders), pastes, crystals, or solids. The target agent can be delivered in any suitable composition, such as, but not limited to, liquids or solutions (e.g., when the target agent is a cell type, the cells can be delivered in a cell suspension), gels (e.g., hydrogels or sol-gels), powders, solids, etc. Therefore, useful material transfer devices and methods of use are disclosed herein.

[0049] Material transfer systems

[0050] In one aspect, disclosed herein is a material (e.g., fluid) transfer system that facilitates simultaneous transfer of material (e.g., fluid) to and / or from pores of a porous container (e.g., a plate), such as by using gravity and / or centrifugal force (e.g., centrifugal force generated by a centrifuge). In one aspect, the configuration of the transfer system at least partially determines the flow path of the fluid transferred to and / or from the pores of the container. In another aspect, the configuration of the transfer system at least partially determines the magnitude of the force required to generate a large amount of fluid flow between at least two containers. In order to regulate the transfer of the fluid, the porous container (e.g., a plate) can be reversibly coupled to at least one other container, such as another porous container (e.g., a plate), that serves as a receiving container. For example, Figure 1A A transfer system 100 is shown, which includes a supply plate 104, a transfer adapter 106, and a receiving plate 108, which can be reversibly coupled to form a transfer assembly 102, such as Figure 1B shown.

[0051] In one aspect, a supply container (e.g., a plate) includes a plurality of wells, at least some of which include one or more target agents. Figure 1A As shown, the supply plate 104 is a multi-well plate including a plurality of wells 110, at least some of which may include one or more target agents and / or be at least partially filled with a fluid. In one aspect, the wells 110 containing at least one target agent may be at least partially filled with a fluid, and the systems and methods described herein are configured to facilitate the transfer of the fluid to isolate at least one target agent and / or fluid. In one aspect, the transfer adapter 106 includes a plurality of transfer lumens 112, which may extend from a first side 114 to a second side 116 of the transfer adapter. In one aspect, as shown in FIG. Figure 1A As shown, the receiving plate 108 includes a plurality of receiving holes 118. In one aspect, the receiving plate can have the same structure as the supply plate, but when the transfer assembly is initially formed, the receiving holes can be empty (e.g., they may not contain a target agent or fluid). In some embodiments, the three components of the transfer assembly (e.g., 104, 106, and 108) are configured such that when the transfer assembly (e.g., 102) is formed, each hole of the supply container (e.g., plate) can be aligned with a different transfer lumen (e.g., 112) of the transfer adapter and a different receiving hole (e.g., 118) of the receiving container (e.g., plate). In some embodiments, the transfer assembly is inverted to a position where the supply container is above the transfer adapter and the receiving container. For example, when the supply plate 104 is above the transfer adapter 106 and the receiving container 108, the fluid can flow from the hole 110 through the transfer lumen 112 and into the receiving hole 118.

[0052] Alternatively, the receiving container (e.g., a plate) can include a plurality of wells, at least some of which include one or more target agents. For example, the receiving plate 108 can be a multi-well plate including a plurality of wells 118, at least some of which can include at least one target agent. In one aspect, the systems and methods described herein are configured to facilitate transfer of fluids to the receiving wells. In one aspect, the transfer adapter 106 includes a plurality of transfer lumens 112, which can extend from a first side 114 of the transfer adapter to a second side 116. In one aspect, the supply plate 104 includes a plurality of supply wells 110. As shown in FIG. Figure 1A As shown, the receiving plate can have the same structure as the supply plate 104, but the receiving wells do not contain fluid when the transfer assembly 102 is initially formed. In some embodiments, the three components of the transfer assembly (e.g., 104, 106, and 108) are configured so that when the transfer assembly (e.g., 102) is formed, each well of the supply container (e.g., plate) can be aligned with a different transfer lumen (e.g., 112) of the transfer adapter and a different receiving well (e.g., 118) of the receiving container (e.g., plate). In some embodiments, the transfer assembly is inverted to a position where the supply container is above the transfer adapter and the receiving container. For example, when the supply plate 104 is above the transfer adapter 106 and the receiving container 108, the fluid can flow from the well 110 through the transfer lumen 112 and into the receiving well 118 (e.g., to contact and / or react with one or more target agents in one or more receiving wells).

[0053] In one aspect, the supply container is configured to removably and / or sealingly couple with a first side of the transfer adapter, and the receiving plate is configured to removably and / or sealingly couple with a second side of the transfer adapter. Figure 1A As shown, the supply plate 104 is configured to be removably coupled to the first side 114 of the transfer adapter 106, and the receiving plate 108 is configured to be removably coupled to the second side 116 of the transfer adapter. On the one hand, in addition to the alignment hole 110, the transfer lumen 112, and the receiving hole 118, the connection of the three components (e.g., 104, 106, and 108) seals the supply hole against the first side of the transfer adapter and seals the receiving hole against the second side of the transfer adapter. In some embodiments, the seal is configured to prevent, minimize, or reduce mixing of contents (e.g., mixing of fluids) between the supply holes or between the receiving holes. For example, the sealing of the supply holes against the first side of the transfer adapter can prevent, minimize, or reduce mixing between the supply holes, such as mixing between two adjacent supply holes. Similarly, the sealing of the receiving hole against the second side of the transfer adapter can prevent, minimize, or reduce mixing between the receiving holes, such as mixing between two adjacent receiving holes. When the supply container, transfer adapter, and receiving container are aligned, the seal allows contents to be transferred between the supply aperture and its corresponding receiving aperture (and no other receiving apertures) while preventing, minimizing, or reducing mixing between the supply aperture and other receiving apertures.

[0054] In some embodiments, the supply plate, transfer adapter, and / or receiving plate include one or more features configured to facilitate alignment of plate structures (e.g., supply holes, transfer lumens, and receiving holes), coupling of components, and / or sealing of components. Figure 1A The transfer adapter 106 shown in FIG. 1 may include a primary extension 120 configured to be inserted into the supply hole 110 of the supply plate 104, and a secondary extension 122 configured to be inserted into the receiving hole 118 of the receiving plate 108. Inserting the primary extension and the secondary extension into the supply hole and the receiving hole, respectively, may facilitate alignment and / or sealing.

[0055] In some embodiments, the receiving plate and the supply plate may have different structures. In one aspect, the supply plate includes a boundary wall, a guide hole structure, and an optional separation hole structure. Figure 1C In the transfer system 150 shown, the supply plate 154 includes a boundary wall 160, a guide hole structure 162 and a separation hole structure 164, and the receiving plate 158 is a multi-hole plate. In some embodiments, the supply plate and the transfer adapter (e.g., Figure 1C In some embodiments, the feed plate includes a separation hole structure coupled thereto. In some embodiments, the separation hole structure is configured to be removably coupled to the feed plate, and assembling the transfer assembly further includes coupling the separation hole structure to the feed plate. In some embodiments, the separation hole structure is fixedly attached or integrated with the feed plate.

[0056] See also Figure 1D As shown in the transfer assembly 152, the transfer assembly can be assembled in a first position, wherein the transfer adapter 156 is positioned below the supply plate 154 and above the receiving plate 158. In one aspect, the supply plate 154 is in an upright position, wherein the top openings of the holes face upward, and the bottom openings of the holes (not shown) face downward and toward the transfer adapter 156. In another aspect, the receiving plate 158 is in an upright position, wherein the inlet of the receiving hole faces upward toward the transfer adapter 156. In this position, the fluid can be retained in the hole of the supply plate due to the properties of the transfer adapter and / or the fluid, and can be transferred into the receiving hole due to gravity and / or centrifugal force (e.g., centrifugal force generated by a centrifuge). As shown in FIG. Figure 1D As shown, a separation hole structure 164 may be used in conjunction with a boundary wall and guide hole structure to form a separate feed hole.

[0057] Supply plate

[0058] A variety of supply plates may be applicable to transfer systems and components as described herein. However, no matter how the specific type of the supply plate, one of its functions may be to include one or more fluids and optional one or more target agents in one or more separate holes. In certain embodiments, the supply plate may be a porous plate, also referred to as a microtiter plate or a microwell plate. In a particular embodiment, the supply plate may be a standard ready-made porous plate, such as those commonly used in a wide range of laboratory applications. In other embodiments, the supply plate may include one or more custom features, such as those features that facilitate fluid transfer and / or the interaction between the supply plate and the transfer adapter. Although the characteristics of the supply plate may be different (e.g., the number of holes, the size of the hole and the contents of the hole, etc.), any porous plate that can be placed in a centrifuge is applicable to transfer systems as described herein.

[0059] In one aspect, the supply and / or receiving containers described herein can be multiwell plates. Multiwell plates can include a grid of small open holes or wells that can contain reagents or substances of interest. Figure 2 A representative well (200) of a multi-well plate is shown. In some embodiments, a well (e.g., 200) may include an opening (e.g., 202) through a top surface (e.g., 204) of the multi-well plate, and material may enter and / or exit the well through the opening. In some embodiments, a well may include and be defined by a base (e.g., 206) and one or more sides or walls (e.g., 208). In some embodiments, for example, Figure 2 As shown, the cross-sectional shape of the hole is circular. It should be appreciated that the hole can have any suitable cross-sectional shape, such as a triangle, a rectangle, any other quadrilateral, a pentagon, a hexagon, a circle, an ellipse, an oval, any other circular or irregular cross-sectional shape. In some embodiments, one or more walls (e.g., 208) of the hole (e.g., 200) are perpendicular to the top surface (e.g., 204) of the multi-well plate, and / or parallel to the longitudinal axis of the hole, such as Figure 2 As shown. In other embodiments, one or more walls may be angled relative to the longitudinal axis of the hole. In some embodiments, the cross-sectional shape and / or area of ​​the opening (e.g., 202) may be different from the cross-sectional shape and / or area of ​​the base (e.g., 206). In some embodiments, the base includes a flat surface, or includes a concave surface, a convex surface, or any other suitable shape of surface.

[0060] In one embodiment, supply container and / or receiving container can have the hole of any suitable number, for example, at least one hole, or at least two holes in the case of perforated plate.For example, ready-made perforated plate can comprise at least about 6 holes, at least about 12 holes, at least about 24 holes, at least about 48 holes, at least about 96 holes, at least about 384 holes, at least about 480 holes, at least about 1536 holes, at least about 3456 holes, or more than 3456 holes.Hole can have any suitable arrangement, as row and column, and in some embodiments, the distance between every pair of adjacent holes can be roughly the same.In certain embodiments, perforated plate can comprise one or more features that can allow hole to separate and / or combine. For example, in some embodiments, the walls of the wells may be removable and / or repositionable to change the number, shape and / or size of the wells, for example, as described in more detail in international patent application serial number PCT / US15 / 16435, entitled "Multi-well Separation Equipment and Reagent Delivery Device" and filed on February 18, 2015, the contents of which are hereby incorporated by reference in their entirety.

[0061] In some embodiments, the supply plate includes boundary walls that form lateral portions of the holding cavity. Figure 3A As shown, the supply plate 300 includes a boundary wall 302, which forms a side portion surrounding the holding cavity 350. Figure 3A and 3B In the example shown, the boundary wall 302 includes four orthogonal parts: a first part 302a, a second part 302b, a third part 302c, and a fourth part 302d. The four parts can define a rectangular area. In some embodiments, the four orthogonal parts can be a single component, while in other embodiments, the four orthogonal parts can include more than one component (e.g., two, three, four or more), which can be attached or assembled in any suitable manner, such as using an adhesive (e.g., glue, adhesive polymer, etc.), welding, mechanical fasteners, chemical bonding, etc.

[0062] However, it should be appreciated that the boundary wall does not necessarily define a rectangular area, and in addition it does not necessarily include four parts. In some embodiments, for example, the boundary wall may define any polygon, such as a triangle, a quadrilateral (e.g., a parallelogram or a trapezoid), a pentagon, a hexagon, etc. It should be appreciated that the boundary wall is not necessarily substantially flat, and can be bent to define an area with a curved shape, such as a circle, an ellipse, an oval, an annulus, a circular segment, etc. In some embodiments, the boundary wall includes less than four parts (e.g., one, two or three parts) or more than four parts (e.g., five, six, seven, eight or more parts). In other embodiments, the boundary wall defines more than one area. For example, in some embodiments, the boundary wall may include a fifth part, which may be attached to the relative part of the boundary wall, such as being attached to the first part 302a at the first end, and being attached to the third part 302c at the second end. In such an embodiment, the boundary wall may define two rectangular areas.

[0063] In some embodiments, the supply plate further comprises a boundary seal. Figure 3A As shown, the supply plate 300 includes a boundary seal 304. In one aspect, the boundary seal is configured to form a leak-proof seal between the boundary wall and the transfer adapter. For example, when the boundary wall 302 and the transfer adapter 700 are Figure 3A and 3B When coupled as shown, the boundary seal can form a leak-proof seal between the boundary wall and the transfer adapter. The boundary seal can include any suitable material for forming a seal, such as but not limited to rubber, plastic, polymer, or any combination thereof. In some embodiments, the boundary seal includes a suitable material having a shape corresponding to the distal side of the boundary wall. For example, as shown in FIG. Figure 3B As shown, boundary seal 304 may include a thin strip of suitable material having a shape corresponding to distal side 306 of boundary wall 302 .

[0064] In some embodiments, the boundary seal is configured to be located between the boundary wall and the transfer adapter when the boundary wall and the transfer adapter are coupled. In some embodiments, the boundary seal is secured to the distal side of the boundary wall. In these embodiments, the boundary seal can be secured to the distal side in any suitable manner (e.g., such as Figure 3B 306), for example but not limited to, using adhesives (such as glue, adhesive polymers, etc.), welding, mechanical fasteners, chemical bonding, etc. in any suitable combination. In some embodiments, the fixation of the boundary seal to the distal side of the boundary wall forms a leak-proof seal between the boundary seal and the boundary wall, and the compression force between the boundary wall and the transfer adapter can press the boundary seal and the transfer adapter together, thereby also forming a leak-proof seal between the boundary seal and the transfer adapter.

[0065] In other embodiments, the boundary seal is also secured to the first planar surface of the transfer adapter in any suitable manner, such as by using adhesives (e.g., glue, adhesive polymers, etc.), welding, mechanical fasteners, chemical bonding, etc. in any suitable combination. In one aspect, the boundary seal is secured to the first planar surface of the transfer adapter (e.g., Fig. 7A The fixation of the boundary seal (704) as shown in the figure creates a leak-proof seal between the boundary seal and the transfer adapter, while the compression force between the boundary wall and the transfer adapter presses the boundary wall and the boundary seal together, thereby also forming a leak-proof seal between the boundary wall and the boundary seal.

[0066] In still other embodiments, when the boundary wall and the transfer adapter are coupled, the boundary seal is not fixed to the boundary wall or the transfer adapter, but is instead sandwiched between the boundary wall and the transfer adapter by a compressive force. In still other embodiments where the boundary wall is fixedly attached to the transfer adapter, the boundary seal is fixed to both the first planar surface of the transfer adapter and the distal side of the boundary wall.

[0067] However, it should be appreciated that the supply plate described herein need not include a boundary seal. For example, if the boundary wall and substrate are configured to form a holding cavity in which a composition (e.g., a cell suspension) can be properly retained without leakage without a boundary seal, then a boundary seal may be unnecessary. For example, in an embodiment in which the boundary wall is fixedly attached or integrated with a transfer adapter, the supply plate may not include a boundary seal. As another example, in an embodiment in which the boundary wall and the transfer adapter are not fixedly attached or integrated but are configured to form a leakproof seal, the supply plate may not include a boundary seal. For example, if the boundary wall includes a material such as rubber, plastic, or polymer (e.g., an elastic polymer) that is configured to seal or cooperate with a structure or material (e.g., rubber, plastic, or polymer) of the transfer adapter, this may be the case. In these cases, the compression force that presses the boundary wall and the transfer adapter together can directly form a leakproof seal (e.g., without a boundary seal structure) between the boundary wall and the transfer adapter.

[0068] Back to Figure 3A and 3B In embodiments of the supply plate 300 of the embodiment of the transfer adapter 302, the transfer adapter and the boundary wall 302, when coupled, can form a holding cavity 350. In some embodiments, the holding cavity provides an area configured to hold a fluid (e.g., an aqueous composition, etc.). It should be appreciated that in embodiments where the boundary wall is fixedly attached or integral with the transfer adapter, a similar holding cavity can be formed by the boundary wall and the transfer adapter.

[0069] although Figure 3A and 3BThe coupled boundary wall 302 and transfer adapter 700 are shown as forming a single integral holding cavity 350, but in other embodiments, the coupled boundary wall and transfer adapter may form a holding cavity having more than one region. For example, in the above embodiment where the boundary wall includes a fifth portion that can be attached to opposing portions of the boundary wall (e.g., attached to the first portion 302a at a first end and attached to the third portion 302c at a second end), the holding cavity may include two rectangular regions. The two regions may be separated by the boundary wall so that a composition located in one region will not be able to move to another region.

[0070] The holding cavity and the components that form it in turn may have any suitable dimensions. In some embodiments, the supply plate (e.g., Figure 3A and 3B 300 shown in Figure 300 is configured to approximate a standard 96-well plate. Therefore, the length and width of the transfer adapter 700 and the boundary wall 302 can be less than about 13 cm, or about 130 mm×85 mm in their maximum dimension. In some embodiments, the depth of the holding cavity 350 (and therefore the approximate height of the boundary wall 302) can be about 1 mm, about 3 mm, about 5 mm, about 7 mm, about 9 mm, about 11 mm, about 13 mm, about 15 mm, about 17 mm, about 19 mm, about 25 mm, about 30 mm, about 35 mm, about 40 mm, or greater than about 40 mm.

[0071] In one aspect, a separation hole structure is provided herein that is configured to be placed within a holding cavity of a feed plate. In some embodiments, the feed plate includes a separation hole structure that is removably placed or irreversibly disposed within the holding cavity. Figure 4A A separation hole structure 400 configured to be positioned within a holding cavity is shown. Figure 3BThe supply plate 300 in the supply plate may include a separation hole structure 400 that can be placed in its holding chamber 350. In some embodiments, the separation hole structure is configured to be removably connected in the holding chamber. In some embodiments, the separation hole structure is fixedly attached or integrated with the boundary wall (e.g., 302) and / or the transfer adapter (e.g., 700). When the separation hole structure is placed in the holding chamber of the supply plate, it can divide the fluid (e.g., aqueous composition) in the holding chamber into the separated volume / region. This method of dividing the composition into the separated volume / region may be simpler and / or more effective than the traditional method (such as pipetting a certain volume of liquid into a separate hole alone), and the traditional method may require repeated transfer of the composition, rather than transferring the composition into the holding chamber at one time. Once the material is divided into the separated volume / region, this allows the separated volume / region to undergo different processing processes (e.g., by introducing different reagents into each volume / region). In addition, the separation hole structure can also be removed from the holding chamber to allow the contents of the separated volume / region to be modified in batches. When it is desired to perform the same process (e.g., a wash step) on each separate volume / area, this may be simpler and more efficient than performing the process on each separate volume / area individually.

[0072] like Figure 4A As shown, the separation hole structure 400 may include a plurality of separation walls 402. In some embodiments, the separation walls 402 may include two sets of orthogonal parallel walls and may form a grid-like or lattice-like structure such that they form an array or matrix of openings 404 separated by the separation walls 402. When the separation hole structure is coupled in a holding cavity, it may divide the holding cavity into a plurality of separation holes, such as Figure 4A In some embodiments, when the separation hole 408 is Figure 4C When the separation hole structure 400 shown in FIG. 1 is placed in the holding cavity, the boundary wall 302 forms at least a portion of the outer wall of the outermost separation hole 408, such as Figure 4A In other embodiments, as shown in Figure 4C The separation hole structure 400 shown may have its own outer wall, so that when the separation hole structure is placed into the holding cavity, the boundary wall does not need to form the outer wall of the outermost separation hole.

[0073] In some embodiments, the separation hole structure further includes one or more separation seals, such as Figure 4B 406 shown in . Figure 4C In the illustrated embodiment, the separation hole structure 400 may include one or more separation hole lips 412 that are configured to be coupled to the top edge of the boundary wall 302. In some embodiments, the supply plate with separation walls is configured to approximate a standard 96-well plate. Figure 4C (as well as Figure 1C and 1D), the separation wall creates 96 openings (ie, 8 rows x 12 columns) when placed in the holding cavity.

[0074] In some embodiments, the separation wall includes one or more separation wall grooves configured to fluidically connect at least two (e.g., all) of the separation holes. Figure 4D As shown, the separation wall 402 may include one or more separation wall grooves 440 so that some or all of the separation holes 408 can be fluidically connected. On the one hand, the separation wall 402 may include a separation wall groove (e.g., 440) between each pair of adjacent separation holes 408. On the other hand, at least a portion of the separation wall may not contact the transfer adapter, leaving one or more gaps between the transfer adapter and the bottom surface of the separation wall, so that some or all of the separation holes are fluidically connected. In some embodiments, the size of the gap between the transfer adapter and the bottom surface of the separation wall may be less than the height of the fluid in the holding chamber, so that when the separation hole structure is connected in the holding chamber, the separation wall is at least partially immersed in the fluid. On the one hand, the separation wall 402 and the optional boundary wall 302 can form the sidewalls of the separation hole 408, and the transfer adapter can form the base of the separation hole. On the other hand, the contents of the holding chamber 350 can be separated and divided into the separation hole 408. The separation hole structure may include any suitable one or more materials, such as but not limited to rubber, plastic, silicon, ceramic, metal, polymer, glass, etc., or any suitable combination thereof.

[0075] In certain embodiments, when separating wall is connected in the holding chamber, separating wall has the height roughly the same as boundary wall.For example, the height of separating wall and boundary wall can be about 1mm, about 3mm, about 5mm, about 7mm, about 9mm, about 11mm, about 13mm, about 15mm, about 17mm, about 19mm, about 25mm, about 30mm, about 35mm, about 40mm or higher.In certain embodiments, separating wall has the height lower than boundary wall, as long as the height of separating wall (and therefore the depth of separating hole) is greater than the depth of the fluid in the holding chamber when separating wall is connected in the chamber, so as to prevent content from flowing between separating holes above separating wall.In other embodiments, separating wall has the height higher than boundary wall, as long as the height of separating wall (and the height of optional boundary wall) is greater than the depth of the fluid in the holding chamber when separating wall is connected in the chamber.It should be noted that not all openings of separating wall need to be used for material transfer.

[0076] In some embodiments, the separation hole structure substantially fills the holding cavity. For example, the cross-sectional dimensions of the separation hole structure 400 may be substantially the same as the cross-sectional dimensions of the holding cavity 350. In one aspect, the holding cavity 350 is configured to properly accommodate the separation hole structure 400. In other embodiments, the separation hole structure does not necessarily fill the holding cavity. For example, the separation hole structure 400 may have a smaller cross-sectional area than the holding cavity 350, and thus only a portion of the holding cavity 350 may be subdivided into the separation holes 408.

[0077] In some embodiments, one or more of the separating walls include one or more separating wall slots. In one aspect, the separating wall slots include openings in the separating wall. In another aspect, the separating wall slots are located at the bottom of the separating wall. In another aspect, the separating wall slots include gaps in the bottom edge of the separating wall. For example, Figure 4D As shown, one or more separation walls 402 include one or more separation wall grooves 440, such as the opening in the separation wall. In certain embodiments, separation wall grooves 440 are positioned at the bottom of the separation wall, and may include the gap in the bottom edge of the separation wall 402. It should be appreciated that separation wall grooves 440 may be positioned at any position along the separation wall 402, such as inside the separation wall 402 and all on all sides are limited by the separation wall, or at the top of the separation wall, and are the gaps in the top edge of the separation wall. In certain embodiments, separation wall grooves 440 may extend beyond the separation walls 402 that intersect. In other embodiments, separation wall grooves 440 may not extend beyond the separation walls 402 that intersect. In certain embodiments, separation wall grooves 440 may extend beyond one or more separation walls 402 that intersect. In other embodiments, separation wall grooves 440 may not extend beyond one or more separation walls 402 that intersect. In certain embodiments, separation wall grooves 440 may extend along the entire length of the separation wall 402. In other embodiments, separation wall grooves 440 may not extend along the entire length of the separation wall 402.

[0078] In some embodiments, one or more separation walls include one or more separation wall grooves configured to divide the retaining cavity into subsets of separation holes, which are configured to be fluidically connected to other separation holes in the subset, but are not configured to be fluidically connected to separation holes in other subsets. For example, in some embodiments, the first separation wall 402 that bisects the retaining cavity does not include any separation wall grooves 440, and the remaining separation walls 402 include separation wall grooves 440 between each adjacent separation hole 408, resulting in a first subset of separation holes 408 on one side of the first separation wall that are configured to be fluidically connected to each other, and a second subset of separation holes 408 on the other side of the first separation wall that are configured to be fluidically connected to each other, while the first subset of separation holes is not configured to be fluidically connected to the second subset of separation holes. In some embodiments, the separation holes can be subdivided into any number and / or configuration of separation hole subsets that are configured to be fluidly connected only to other separation holes in the same subset.

[0079] In certain embodiments, the separating wall groove can have the cross-sectional area of ​​any suitable shape and / or size. In certain embodiments, the maximum dimension of the cross section of the separating wall groove can be about 1 μm to about 20 μm, about 20 μm to about 40 μm, about 40 μm to about 60 μm, about 60 μm to about 80 μm, about 80 μm to about 100 μm, about 100 μm to about 200 μm, about 200 μm to about 400 μm, about 400 μm to about 600 μm, about 600 μm to about 800 μm, about 800 μm to about 1 mm, about 1 mm to about 2 mm, about 2 mm to about 4 mm, about 4 mm to about 6 mm, about 6 mm to about 8 mm, about 8 mm to about 1 cm, greater than about 1 cm, about 1 μm to about 1 cm, about 100 μm to about 1 mm, or about 1 mm to about 1 cm. However, it should be recognized that in some embodiments, it may be desirable for the cross-sectional area of ​​the separation wall groove to be a particular value in order to allow fluid within the retention chamber to flow freely between the separation wall groove (e.g., 440) and the separation hole (e.g., 408).

[0080] In one aspect, the separation wall grooves can have any suitable shape, such as but not limited to having a cross-section having a triangle, square, rectangle, any other quadrilateral (parallelogram or trapezoid, etc.), pentagon, hexagon, etc., any circular shape (circular, elliptical or oval, etc.) or irregular shape. In some embodiments, one or more separation wall grooves have a rectangular cross-sectional shape, such as Figure 4D In some embodiments, one or more or all of the separating wall grooves have equal cross-sectional size and / or shape. In some embodiments, the separating wall grooves do not necessarily have the same size and / or shape.

[0081] On the one hand, the opening (e.g., 404) may have any suitable cross-sectional area. In some embodiments, the maximum dimension of the cross section of the opening may be about 1 μm to about 20 μm, about 20 μm to about 40 μm, about 40 μm to about 60 μm, about 60 μm to about 80 μm, about 80 μm to about 100 μm, about 100 μm to about 200 μm, about 200 μm to about 400 μm, about 400 μm to about 600 μm, about 600 μm to about 800 μm, about 800 μm to about 1 mm, about 1 mm to about 2 mm, about 2 mm to about 4 mm, about 4 mm to about 6 mm, about 6 mm to about 8 mm, about 8 mm to about 1 cm, greater than about 1 cm, about 1 μm to about 1 cm, about 100 μm to about 1 mm, or about 1 mm to about 1 cm. However, it should be appreciated that in some embodiments, it is desirable that the ratio between the height and the cross-sectional area of ​​the opening (e.g., 404) has a specific value in order to offset the capillary effect. The resulting separation well (e.g., 408) can have any suitable volume, such as, but not limited to, less than about 100 μL, about 100 μL to about 200 μL, about 200 μL to about 400 μL, about 400 μL to about 600 μL, about 600 μL to about 800 μL, about 800 μL to about 1 mL, about 1 mL to about 10 mL, about 10 mL to about 20 mL, about 20 mL to about 40 mL, about 40 mL to about 60 mL, about 60 mL to about 80 mL, about 80 mL to about 100 mL, greater than about 100 mL, about 100 μL to about 100 mL, or about 1 mL to about 10 mL.

[0082] Although the opening (e.g., 404) is Figures 4A-4C 402. In some embodiments, one or more or all of the openings have equal cross-sectional sizes and / or shapes. In some embodiments, the openings do not necessarily have the same size and / or shape. In other embodiments, the cross-sectional area of ​​the openings along the size of the openings is not necessarily the same. For example, in some embodiments, the cross-sectional area of ​​each opening at its proximal end (e.g., close to the transfer adapter) may be greater than the cross-sectional area at its distal end (e.g., away from the transfer adapter). In other embodiments, the cross-sectional area of ​​each opening at its distal end may be greater than the cross-sectional area at its proximal end. It should be recognized that in these cases, the thickness of the separation wall (e.g., 402) may vary accordingly to produce different cross-sectional areas. For example, the thickness of the separation hole at the distal end (e.g., away from the transfer adapter) may be greater than the proximal end (e.g., close to the transfer adapter), or alternatively, the thickness may be greater at the proximal end than at the distal end.

[0083] exist Figure 4A-4B In the illustrated embodiment, the separation hole structure 400 defines 64 openings 404, and thus when coupled within a holding cavity (e.g., 350), the separation hole structure can divide the holding cavity into 64 separation holes 408. However, it should be appreciated that the separation hole structure can define any number of openings, and thus when coupled with a holding cavity (e.g., 350), the separation hole structure can divide the holding cavity into any number of separation holes. For example, the separation hole structure can define at least about 6, at least about 12, at least about 24, at least about 48, at least about 96, at least about 384, at least about 480, at least about 1536, at least about 3456 or more openings, and thus when connected to the holding chamber (e.g., 350), the holding chamber can be divided into at least about 6, at least about 12, at least about 24, at least about 48, at least about 96, at least about 384, at least about 480, at least about 1536, at least about 3456 or more separation holes. It should be understood that the number of openings (e.g., 404) and separation holes (e.g., 408) is not necessarily limited by the number listed here. Although in some embodiments, the separation hole structure can be limited to the number of openings present in a standard microtiter laboratory plate, in other embodiments, the separation hole structure can define a non-standard number of openings, which may or may not be a rectangular number.

[0084] In some embodiments, it is desirable to maximize the number of separation holes (e.g., 408) within a given cross-sectional area (e.g., the cross-sectional area of ​​the holding chamber 350). In order to do so, it may be desirable to minimize the thickness of the separation wall (e.g., 402). In some embodiments, the thickness of the separation wall may be from about 50 μm to about 2000 μm. In particular embodiments, the thickness of the separation wall is from about 200 μm to about 1800 μm, from about 400 μm to about 1600 μm, from about 600 μm to about 1400 μm, from about 800 μm to about 1400 μm, or from about 1000 μm to about 1200 μm.

[0085] It should also be appreciated that in some embodiments, the boundary walls (eg, 302 ) described herein may be used without a separate aperture structure such as 402 .

[0086] In some embodiments, the separation hole structure further comprises a separation seal. On the one hand, when the separation hole structure and the transfer adapter are connected, the separation seal can create a leak-proof seal at the distal end of one or more separation holes. In some embodiments, this can allow each separation hole to become an isolated area so that it can undergo a different process or treatment than one or more separation holes adjacent to it. Figure 4A-4BAs shown, when the separation hole structure and the transfer adapter are coupled, the separation seal 406 can be located between the distal surface of the separation wall 402 of the separation hole structure 400 and the first flat surface (e.g., 704) of the transfer adapter. In some embodiments, the separation seal can cover the bottom edge of at least one or each separation hole, and thus can provide a seal for at least one or each separation hole when pressed onto the transfer adapter. The separation seal can include any suitable material for forming a seal, such as but not limited to rubber, plastic or polymer, or any suitable combination thereof.

[0087] exist Figure 4A-4B In the illustrated embodiment, the separation seal 406 is coupled to the distal surface of the separation wall 402 of the separation hole structure 400. The separation seal can be attached to the separation hole structure in any suitable manner, such as but not limited to, by using an adhesive (e.g., glue, adhesive polymer, etc.), chemical bonding, etc., or any combination thereof. However, it should be appreciated that in other embodiments, the separation seal can be attached to the first flat surface (e.g., 704) of the transfer adapter. In yet other embodiments, the separation seal can be attached to the boundary wall (e.g., 302). In some embodiments, the separation seal can be integral with the boundary seal (e.g., 304), or it can be attached to the boundary seal.

[0088] It should also be recognized that the separation hole structure (e.g., 400) does not necessarily include a separation seal (e.g., 406). If the separation hole structure and the transfer adapter are configured to form a separation hole that can properly retain a fluid (e.g., an aqueous composition) therein without leakage in the absence of a separation seal, then a separation seal may be unnecessary. For example, if the separation seal includes a material that can form a seal with the material of the transfer adapter, such as rubber, plastic, or polymer, this may be the case. In these cases, the compression force that presses the separation hole structure and the transfer adapter together can directly form a leak-proof seal between the separation hole structure and the transfer adapter without the need for an intermediate separation seal. As another example, as described in more detail below, the supply plate may not include a separation seal in some (but not all) embodiments in which it includes a guide hole structure.

[0089] In some embodiments, the separation hole structure (e.g., 400) can be configured to be coupled within the holding cavity (e.g., 305) such that when coupled, sufficient compressive pressure exists between the separation hole structure and the transfer adapter (e.g., 700) such that a leak-proof separation hole (e.g., 408) is formed. In some embodiments, the separation hole structure can be configured to be coupled within the holding cavity via the boundary wall (e.g., 302) and / or the transfer adapter (e.g., 700). Figure 4A-4B In the illustrated embodiment, the separation hole structure 400 includes a separation hole clip 410 configured to be coupled to the boundary wall 302. Figure 4C In the illustrated embodiment, the separation hole structure 400 includes a separation hole lip 412 configured to be coupled to a top edge of the boundary wall 302 .

[0090] In some embodiments, the supply plate (e.g., 300) includes a separation hole structure (e.g., 400) that is configured to be removably coupled within a holding cavity (e.g., 350). In one aspect, the design of the coupling mechanism between the separation hole structure and the holding cavity can allow the separation hole structure to be removed from the holding cavity after the two elements have been coupled. The ability to separate and remove from the holding cavity can allow the separation hole structure to be inserted to first separate the fluid in the holding cavity into the separation hole, and then can allow the separation hole structure to be removed to recombine the fluid in the separation hole.

[0091] In some embodiments, the feed plate (e.g., 300) includes a separation hole structure (e.g., 400) fixedly coupled within a holding cavity (e.g., 350). In one aspect, the design of the coupling mechanism between the separation hole structure and the holding cavity can be such that the separation hole structure cannot be removed from the holding cavity after the two elements have been coupled. In some embodiments, the feed plate (e.g., 300) includes a separation hole structure (e.g., 400) fixedly attached or integral with a boundary wall (e.g., 302) and / or a transfer adapter (e.g., 700).

[0092] It should be appreciated that any of the separation pore structural features described may be combined in any suitable manner. For example, an embodiment of a transfer adapter may include a transfer adapter from reference Figure 4A-4B Some components of the embodiments discussed and from reference Figure 4C-4D Other components of the discussed embodiments.

[0093] In some embodiments, the supply plate (e.g., 300) includes a guide hole structure (e.g., 502). The guide hole structure may be configured to reduce the cross-sectional area of ​​the distal portion of the separation hole (e.g., 408). This may be advantageous, for example, because it can guide the fluid in each separation hole to a smaller cross-sectional area at the base of the separation hole aligned with the opening in the transfer adapter (e.g., 700). In some embodiments, the guide hole structure may include a thin layer of material, such as but not limited to a soft and / or elastic material (e.g., silicone, rubber, etc.), which may be configured to be located between the separation hole structure and the transfer adapter. Generally, the guide hole structure may include a plurality of openings, which may correspond to the distal ends of the openings of the separation hole structure at the proximal end, and the openings may narrow in the direction from the proximal side to the distal side. The distal end of the opening in the guide hole structure may allow the fluid in the separation hole to be guided to the opening in the transfer adapter. In some embodiments, the center of the separation hole may not be aligned with the opening in the transfer adapter, and the distal end of the opening in the guide hole structure may be positioned to guide the fluid flow in the separation hole to the opening in the transfer adapter.

[0094] Figure 5A-5B A perspective view and a top view of one embodiment of a guide hole structure 502 are shown, respectively. As can be seen therein, the guide hole structure 502 may include a thin structure including a plurality of openings 504. The cross-sectional shape of the openings 504 at the proximal side may be configured to correspond to the cross-sectional shape of the separation hole 408. The openings 504 may have an inverted truncated quadrangular pyramid shape such that the cross-sectional area of ​​the openings 504 decreases from proximal to distal. At the distal end of the openings 504, a portion of the transfer adapter 700 may be exposed.

[0095] Figure 5A-5B 502. A perspective view and a top view of one embodiment of a guide hole structure are shown, respectively. In these examples, the guide hole structure 502 comprises a thin structure comprising a plurality of openings 504. The cross-sectional shape of the openings 504 at the proximal side may be configured to correspond to the cross-sectional shape of the separation hole 408. The openings 504 may have an inverted truncated quadrangular pyramid shape such that the cross-sectional area of ​​the opening decreases from the proximal end to the distal end. At the distal end of the openings 504, a portion of the transfer adapter 700 may be exposed. Figure 5C A close-up view from one side of a guide hole structure 502 having an opening 504 having an inverted truncated quadrangular pyramid shape is shown. In one aspect, the cross-sectional area of ​​the opening at the proximal end 506 is greater than the cross-sectional area of ​​the opening at the distal end 508. In other embodiments, the opening may have other shapes, such as, but not limited to, a truncated cone or a truncated pyramid.

[0096] In one aspect, the guide hole structure can be located between the separation hole structure and the transfer adapter. In some embodiments where the feed plate includes a guide hole structure, the feed plate may not include a separation seal. For example, the feed plate may include a guide hole structure without a separation seal, and the guide hole structure includes a material that may be able to form a seal between the transfer adapter and the separation hole structure, such as rubber, plastic, or polymer. In some embodiments, the guide hole structure can be attached to a transfer adapter (e.g., Figure 1C 700) shown in ); the guide hole structure can be attached to a first flat surface (e.g., 704) of the guide hole structure; the guide hole structure can be attached to a boundary wall; the guide hole structure can be attached to a boundary seal (e.g., by attaching the outer edge of the guide hole structure to the inner edge of the boundary seal); and / or the guide hole structure can be attached to a separation hole structure. The guide hole structure can be attached to these elements (e.g., the first flat surface, the boundary wall, the boundary seal, and / or the separation hole structure) in any suitable manner, such as but not limited to an adhesive (glue, adhesive polymer, etc.), chemical bonding, etc. In some embodiments, the guide hole structure can be fixedly attached to any of these elements (e.g., the first flat surface, the boundary wall, the boundary seal, and / or the separation hole structure). In some embodiments, the guide hole structure, the transfer adapter, the boundary wall, and / or the separation hole structure are integrally formed.

[0097] In other embodiments where the feed plate includes a guide hole structure, the feed plate may further include a separation seal. For example, the feed plate may include both the guide hole structure and the separation seal in embodiments where the guide hole structure includes a material that is generally not capable of forming a sufficient seal between the transfer adapter, the guide hole structure, and the separation hole, such as glass or hard plastic. In these embodiments, the separation seal may be located between the transfer adapter and the guide hole structure and / or between the guide hole structure and the separation hole structure. When the separation seal is located between the transfer adapter and the guide hole structure, the separation seal may be attached to the first flat surface of the transfer adapter, the distal surface of the guide hole structure, or the boundary wall. When the separation seal is located between the guide hole structure and the separation hole structure, the separation seal may be attached to the proximal surface of the guide hole structure, the distal surface of the separation hole structure, or the boundary wall.

[0098] In some embodiments, the supply plate described herein may further include a cover. The cover may be configured to be assembled over the supply plate to cover the holding cavity. In some embodiments, the cover may be configured to individually seal the top of each separation hole when the separation hole structure is coupled in the holding cavity.

[0099] In some embodiments, at least one or each hole in a plurality of holes may have a volume of about 100 μL to about 100 mL. In some embodiments, at least one or each hole may have a volume of less than about 100 μL, about 100 μL to about 200 μL, about 200 μL to about 400 μL, about 400 μL to about 600 μL, about 600 μL to about 800 μL, about 800 μL to about 1 mL, about 1 mL to about 10 mL, about 10 mL to about 20 mL, about 20 mL to about 40 mL, about 40 mL to about 60 mL, about 60 mL to about 80 mL, about 80 mL to about 100 mL, or a volume greater than about 100 mL. In some embodiments, each hole in a plurality of holes may have a depth of about 1 mm to about 40 mm. In some embodiments, each hole in the plurality of holes may have a depth of about 5 mm to about 15 mm, about 10 mm to about 20 mm, about 15 mm to about 25 mm, about 20 mm to about 30 mm, about 25 mm to about 35 mm, about 30 mm to about 40 mm, or greater than about 40 mm.

[0100] The porous container (e.g., plate) may include any suitable one or more materials, including but not limited to glass, plastic, and metal. In some embodiments, at least a portion of the inner surface of the hole may include a coating, and in some embodiments, only the base may include a coating. The coating may perform one or more useful functions, such as fixing a target agent. For example, the coating may include one or more compounds, proteins, gels (e.g., hydrogels), polymers, copolymers, fixed cells, microorganisms, etc.

[0101] The hole of the supply plate can be at least partially filled with any suitable content. Generally, the content of the supply hole of the supply plate used in the transfer system described herein can include one or more fluids and optional one or more target agents. One or more fluids can at least partially fill the supply hole. The target agent can be at least temporarily fixed on or fixed on the inner surface or base of the supply hole and immersed in the fluid. Through the transfer process, the fluid can be basically transferred from the supply hole, and the target agent can remain fixed on the inner surface of the hole to a large extent. Any suitable target agent can be used, such as protein, nucleic acid, microorganism (e.g., bacteria, fungi), plant (e.g., algae), virus, small molecule drug or any compound, polymer, antigen, antibody, cell fragment, cell homogenate, DNA, peptide, etc. The fluid can include any suitable fluid, such as aqueous fluid.

[0102] A supply plate suitable for use with the material transfer system described herein may include one or more features to facilitate coupling, sealing and / or alignment with a transfer adapter. In some embodiments, at least a portion of the top surface of the supply plate may be covered with an adhesive that may be configured to contact the surface of the transfer adapter. The adhesive may help couple the supply plate and the transfer adapter together and / or form a seal that prevents fluid from flowing between different holes in the supply plate. Additionally or alternatively, the top surface of the supply plate may include a structure that engages a corresponding structure on the surface of the transfer adapter. For example, the supply plate may have a lip around the periphery of the top surface, around the supply hole, or at any other suitable position in a corresponding groove that can be assembled on the transfer adapter. This may help couple, seal and align the supply hole of the supply plate with the opening of the transfer adapter. As other examples, the supply plate may include a clip, a fixture, a latch, etc. that is configured to dock with the transfer adapter. The supply plate may also be configured to dock with a clip, a fixture, a latch, etc. on the transfer adapter. Clips, fixtures, latches, etc. may be attached to the transfer adapter and / or receiving plate to facilitate coupling, sealing and / or alignment of two or more plates.

[0103] Although in some embodiments the supply plate is described as a separate and distinct element from the transfer adapter, it should be appreciated that in some embodiments the supply plate may be integrally formed with the transfer adapter. In these embodiments, the combined supply plate-transfer adapter may be coupled to the receiving plate to facilitate the transfer of material (e.g., fluid) from the holes of the supply plate to the holes of the receiving plate. The combined supply plate-transfer adapter may be configured to allow or prevent mixing of fluids from different holes of the supply plate.

[0104] Transfer Adapter

[0105] On the one hand, the transfer system described herein may include a transfer adapter configured to adjust the transfer of fluid to or from the holes of a porous plate. When the plates of the transfer system are combined to form a transfer assembly, a first side of the transfer adapter may be (reversibly or irreversibly) coupled to a supply plate, and a second opposite side of the transfer adapter may be (such as reversibly) coupled to a receiving plate. In some embodiments, the transfer adapter and the supply plate are formed integrally and the exposed surface of the transfer adapter may be coupled (such as reversibly) to the receiving plate. The transfer adapter may include a flat sheet having a plurality of openings, and the layered arrangement of the transfer assembly may allow the fluid to flow from the holes of the supply plate, through the openings in the transfer adapter, and into the receiving plate. In order to facilitate such fluid flow, the openings in the transfer adapter may generally be at least as many as the holes in the supply plate, and the openings may be arranged so that each opening of the transfer adapter is aligned with a different hole in the supply plate. For example, the distance between the centers of adjacent openings, the arrangement of the openings in rows and columns, and the number of openings in each row and column may match the arrangement of the holes in the supply plate. In some embodiments, the transfer adapter may be configured to align more than one opening with each hole in the supply plate. In embodiments of the transfer assembly that include a receiving plate having a plurality of receiving holes, each opening of the transfer adapter may be aligned with a different receiving hole, or more than one opening may be aligned with each receiving hole.

[0106] The transfer adapter may include one or more features to facilitate alignment of the openings with the holes of the supply plate, and in some embodiments, with the receiving holes of the receiving plate. For example, the transfer adapter may include one or more structures near at least one opening that may act as insertion guides to assist in alignment when the transfer adapter and the supply plate are connected. In some embodiments, these structures may include extensions protruding from the surface of the flat sheet adjacent to or around each opening. Each extension may engage (e.g., contact, enter, at least partially fit into) a different supply hole to align the opening with the supply hole. The transfer adapter may include extensions on a first side to engage the holes of the supply plate and / or on a second side to engage the receiving holes of the receiving plate.

[0107] Additionally or alternatively, the transfer adapter may include an alignment guide that is aligned with a predetermined portion of the supply plate. For example, the alignment guide may be one or more dimensions (e.g., length, width) of the transfer adapter that can match the corresponding dimensions of the supply plate. Aligning the corresponding dimensions of the transfer adapter and the supply plate can also align the opening with the hole. As another example, the alignment guide may include one or more pins and / or notches that are configured to dock with corresponding features on the supply plate. It should be appreciated that the transfer adapter may have the same or different alignment guides that can be aligned with the predetermined portion of the receiving plate so as to align the opening with the receiving hole.

[0108] On the one hand, coupling the transfer adapter to the supply plate and the receiving plate can form a fluid-tight or leak-proof seal. For example, a seal can be formed between each hole of the supply plate and the transfer adapter to prevent fluid flow between the supply holes. In other words, the fluid can be allowed to flow out of the supply holes and through the opening in the transfer adapter, but the seal can prevent the fluid from flowing out of one supply hole and into a different supply hole. For example, when different experimental protocols have been performed in different supply holes, it may be desirable to prevent fluid mixing between the supply holes and therefore to maintain isolation of the hole contents. When the receiving plate includes a plurality of receiving holes, it may be desirable to form a seal between each receiving hole and the transfer adapter so as to isolate the transferred fluid from each hole of the supply plate. Another function of the seal can be to hold the plates of the transfer assembly together by friction so that they do not accidentally separate.

[0109] On the one hand, the transfer adapter may include one or more features to facilitate sealing. For example, in an embodiment of a transfer adapter including an extension, each extension may be fitted into a hole (e.g., a supply hole of a supply plate or a receiving hole of a receiving plate) and seal against the inner surface of the hole. The size and shape of a portion of the extension may match the size and shape of a portion of the hole, and these portions may interact to form a seal. The material properties of the extension and / or the hole may improve the seal (e.g., reduce the chance of fluid leaking through the seal, and / or increase the force required to break the seal). For example, the extension may include one or more compliant materials (e.g., rubber, plastic, or polymer). This may allow the extension to conform to the shape of the hole. The materials of the extension and the hole may also be selected to increase or reduce the friction that holds the extension and the hole together.

[0110] Some embodiments of the transfer adapter may not include extensions or other structures that enter the holes of the supply plate or receiving plate. In these embodiments, the flat surface of the transfer adapter may seal against the top surface of the supply plate around the hole opening. The transfer adapter and / or the supply plate may include one or more features to facilitate the sealing of these surfaces. For example, an adhesive may be on the surface of the transfer adapter and / or the supply plate. Additionally or alternatively, a clamp or fixture may compress the surfaces of the transfer adapter and the supply plate together. It should be recognized that any sealing mechanism described herein that can seal the holes of the supply plate against the transfer adapter can also be used to seal the receiving holes of the receiving plate against the transfer adapter. In some embodiments, the flat surface of the transfer adapter may be fixedly connected to the bottom surface of the supply plate.

[0111] The transfer adapter can regulate the flow of fluid in many ways. For example, alignment and sealing mechanisms can guide the fluid to flow through the opening. In other examples, one or more features of the transfer adapter can determine the size of the force required to cause the fluid to flow through the opening. In some embodiments, the size and / or shape of the opening can make the gravity acting on the fluid in the supply hole positioned above the transfer adapter cause the fluid to flow through the opening. In other embodiments, the size and / or shape of the opening can make it possible for gravity alone to not cause the fluid to flow through the opening. For example, if the opening includes one or more small holes or slits, the cohesive force (e.g., surface tension) of the fluid and the adhesion between the fluid and the transfer adapter may prevent the fluid from flowing. Therefore, the magnitude of the force required to cause the fluid to flow through the opening in large quantities may depend on the size and / or shape of the opening, the characteristics (e.g., hydrophobicity, hydrophilicity) of the material around the opening, and the characteristics (e.g., viscosity) of the fluid. If the transfer adapter is configured so that cohesive force and adhesion prevent the fluid from flowing through the opening simply due to gravity, then applying an external force (such as the force generated by a centrifuge) can produce fluid flow.

[0112] Additionally or alternatively, the transfer adapter may be configured such that an external force (e.g., from centrifugation of the transfer adapter) may change the area of ​​each opening. For example, in some embodiments, one or more leaflets may surround and / or define the area of ​​each opening. In the absence of any external force, the leaflets may be in a closed first position to prevent fluid from flowing through the opening. In this first position, the opening may be completely closed, or the area may be small enough to prevent fluid from flowing due to gravity. When an external force is applied to the transfer adapter (e.g., by centrifuging the transfer adapter), the leaflets may deflect, deform, or otherwise move to a second open position. When the leaflets are in the second open position, the area of ​​the opening may increase, and fluid may be able to flow through the opening. A transfer adapter that allows a large amount of fluid to flow only when an external force is applied may have several advantages. For example, this prevents fluid from accidentally flowing back from the receiving plate to the supply plate after the fluid is transferred. This construction may also allow the transfer adapter to be used as a liquid-tight cover or lid for the supply plate.

[0113] Fig. 6A and 6B An example of a transfer adapter configured for use with a multi-well container (e.g., a 96-well plate) is depicted in FIG. Fig. 6A , the transfer adapter 600 includes a substrate (e.g., a flat sheet 602), a plurality of openings on the substrate, and a primary extension 606 and a secondary extension 608 associated with each opening. In this example, the openings and the associated primary extensions and secondary extensions are arranged to correspond to the wells of a 96-well plate, for example, in 6 rows x 12 columns. However, it should be understood that the openings and the associated primary extensions and secondary extensions can be arranged in any suitable pattern to correspond to some or all of the wells of any multi-well container.

[0114] To better illustrate the opening, Figure 6B A row of openings 604 is depicted without primary and secondary extensions. In one aspect, each opening comprises a through hole extending between a first surface and a second surface of the substrate. For example, Figure 6B As shown, openings 604 are through holes extending between first planar surface 610 and second planar surface 612 of planar sheet 602. In this example, there are 96 openings 604 arranged so that each is aligned with a different well of a 96-well plate when the transfer assembly is formed.

[0115] In one aspect, the transfer adapter includes a main extension that protrudes from the first surface around an opening in the transfer adapter base. Fig. 6A and 6B As shown, the transfer adapter 600 includes a primary extension 606 protruding from a first planar surface 610 around each opening 604. In another aspect, the transfer adapter further includes a secondary extension protruding from a second surface around the opening on the transfer adapter base. Figure 6B As shown, in addition to the primary extension 606 , the transfer adapter 600 further includes a secondary extension 608 that may protrude from the second planar surface 612 around each opening 604 .

[0116] In one aspect, the primary extension includes a primary lumen and / or the secondary extension includes a secondary lumen. Figure 6B As shown, each primary extension 606 includes a primary lumen 618, and each secondary extension 608 includes a secondary lumen 620. In some embodiments, the primary lumen and the secondary lumen are configured to form a continuous transfer lumen extending between the first side and the second side of the transfer adapter. For example, both the primary lumen and the secondary lumen are configured to be fluidly connected to the same opening on the base of the transfer adapter, thereby forming a continuous transfer lumen through the base. Figure 6B As shown, each primary lumen 618 is aligned with and / or configured to be fluidly connected to its corresponding opening 604 and secondary lumen 620, thereby forming a continuous transfer lumen extending between the first side 614 and the second side 616 of the transfer adapter 600. In some embodiments, when the first side of the transfer adapter is coupled to the supply plate, each transfer lumen can be aligned with a different hole of the supply plate to facilitate fluid flow out of the multiple holes, through the multiple transfer lumens, and into the receiving plate.

[0117] In another aspect, the main extension facilitates alignment and / or sealing of the transfer adapter and the supply plate, thereby facilitating fluid flow from the supply plate aperture through the opening of the transfer adapter. In one embodiment, the main extension is configured to at least partially fit into the aperture of the supply plate and seal against the aperture. In a particular embodiment, the main extension is configured to form a seal (e.g., a fluid-tight seal) with an inner surface of the supply plate aperture. For example, Figure 6C and6D As shown, each main extension 606 is configured to at least partially fit into a different hole in the supply plate and seal against the hole (e.g., form a seal with its inner surface). Figure 6C Details of the main extension can be seen in a close-up view of a portion of the first side 614 of the transfer adapter. Fig.6D A cross-sectional view of a main extension 606 inserted into a hole 622 of a multiwell plate is provided in FIG. In some embodiments, the main extension includes an inner end adjacent to the base of the transfer adapter and an outer end distal to the base. For example, as shown in FIG. Fig.6D As shown, each main extension 606 may include an inner end 624 adjacent to the flat sheet 602 and an outer end 626 away from the flat sheet. In one embodiment, the main extension tapers from the inner end to the outer end so that the inner cross-sectional area is larger than the outer cross-sectional area of ​​the main extension. Fig.6D As shown, each main extension 606 tapers from the inner end 624 to the outer end 626, so that the inner cross-sectional area is larger than the outer cross-sectional area. On the one hand, the outer cross-sectional area of ​​the main extension is smaller than the inner cross-sectional area of ​​the hole, so that the outer end of the main extension can be easily inserted into the hole and moved forward into the hole. For example, Fig.6D As shown, when the outer end 626 of the main extension 606 is inserted into the hole 622, the outer cross-sectional area of ​​the outer end of the main extension is smaller than the corresponding inner cross-sectional area of ​​the hole, for example, when the cross-sectional area of ​​the outer end and the cross-sectional area of ​​the hole opening are taken from the same cross-sectional plane. In addition, inserting the outer end 626 of the main extension 606 into the hole 622 can align each main lumen 618 and each opening of the transfer adapter with a different hole of the supply plate.

[0118] In yet another aspect, a sealing area is provided on the outer surface of the main extension, the sealing area being configured to contact and seal against the inner surface of the hole. Fig.6D As shown, the outer end 626 of the main extension 606 can be moved forward into the hole 622 until a portion of the outer surface of the main extension (sealing area 628) contacts and seals against the inner surface of the hole 622. In one aspect, when the seal is formed, the cross-sectional shape and area of ​​the main extension (on its outer surface) are substantially the same as the inner cross-sectional shape and area of ​​the portion of the hole that the sealing area contacts. In one aspect, this enables a continuous seal to be formed between the outer periphery of the main extension and the inner periphery of the hole. Figures 6A-6C In the example shown, the cross-sectional shape of the main extension 606 can be circular to allow it to form a continuous circumferential seal with a hole that also has a circular cross-sectional shape. However, the sealing area of ​​the main extension can have any shape that matches the internal cross-sectional shape of the hole (e.g., square, rectangular, oval, or elliptical). In some embodiments, the main extension is fully inserted into the hole so as to form a fluid-tight seal against the hole at the sealing area. In other embodiments, partially inserting the main extension into the hole is sufficient to form a fluid-tight seal against the hole. For example, although Fig.6D Main extension 606 is depicted fully inserted into bore 622, but it should be appreciated that in some embodiments, a fluid-tight seal may be formed before the main extension is fully within the bore.

[0119] In one aspect, the main extension includes one or more portions having different slopes or angles relative to the transfer adapter base. In some cases, a main extension having two or more slopes may be advantageous over a main extension having one slope. For example, Figure 6C and 6D As shown, the main extension 606 includes at least two portions having different slopes or angles relative to the flat sheet 602. Specifically, the outer surface of the sealing area 628 has a first slope having an angle α relative to the flat sheet 602, and the outer surface of the end of the main extension away from the flat sheet 602 (e.g., the area 632 of the main extension 606) has a second slope having an angle β relative to the flat sheet. In some embodiments, the first slope can be substantially the same as the slope of the inner surface of the hole 622, which can maximize the contact area between the sealing area 628 and the inner surface of the hole. In some embodiments, the wall of the hole 622 is vertical (e.g., perpendicular to the top surface 630 of the multiwell plate), and thus the first slope can be vertical (e.g., perpendicular to the flat sheet 602 of the transfer adapter). In some embodiments, the second slope of the area 632 can be less than the slope of the sealing area 628 (i.e., the angle β can be less than the angle α), and thus the area 632 can not contact and seal against the inner surface of the hole 622. On the one hand, the smaller second slope can result in a smaller cross-sectional area at the outer end 626, thereby facilitating the insertion of the main extension into the hole 622. On the other hand, the smaller slope can reduce the length of the main extension 606, for example, resulting in a shorter longitudinal distance between the inner end 624 and the outer end 626. This configuration can reduce the chance of contact between the main extension and the target agent in the hole (e.g., the target agent 634 at the bottom base of the hole 622).

[0120] On the one hand, if the slope of the distal region of the main extension is less than the slope of the inner surface of the hole, a recess may be formed between the main extension and the inner surface of the hole. Fig.6D A pocket 636 is shown formed between the main extension 606 and the hole 622 because the slope of the region 632 is less than the slope of the inner surface of the hole when the main extension is inserted into the hole. In this example, when the transfer assembly is inverted, or during centrifugation, fluid can enter this pocket 636 rather than flowing directly into the main lumen 618 and then out of the hole 622.

[0121] In one aspect, the main extension includes one or more features to reduce the chance of fluid being trapped between the transfer adapter and the aperture (e.g., between the main extension and the inner surface of the aperture, such as in pocket 636). In some embodiments, the one or more features include features in the structure, size, and / or shape of the main extension (e.g., slits, gaps, notches, orifices, grooves, channels, etc.), and / or features in the material properties of the main extension (e.g., hydrophobicity or hydrophilicity). For example, Figure 6C As shown, each main extension 606 may include one or more slits 638 (e.g., four slits 638) configured to be fluidly connected to the main lumen 618. In one aspect, the slits may be gaps in the periphery of the main extension (e.g., in region 632). In one aspect, fluid entering the recess may flow through the slits or gaps into the main lumen and out of the hole, rather than remaining trapped in the recess. In another aspect, as shown in FIG. Figure 6C As shown, each slit 638 may include an inclined or angled base 640 directed toward the center of the main lumen 618, thereby directing fluid out of the recess 636, into the main lumen, and ultimately out of the aperture 622. Figure 6C A slit is shown in the figure, but it should be appreciated that any structure configured to fluidically connect the recess to the main lumen can reduce the chance of fluid being retained in the recess. In some embodiments, the structure includes an orifice, a hole, a slit, a gap, a notch, a groove, or a channel, etc.

[0122] In some embodiments, the main extension includes a region having an outer slope that is the same as the slope of the inner surface of the hole and an inner slope that is less than the slope of the inner surface of the hole, wherein there are no pockets because the material of the main extension will fill the pockets. Fig.6D , main extension 606 may include a region 632 having an outer slope that is the same as the slope of the inner surface of the bore and an inner slope that is less than the slope of the inner surface of the bore, without a pocket 636. In such a main extension, outer end 626 of main extension 606 may include a surface that is curved or angled (e.g., concave or conical) in a manner to direct fluid from the bore into main lumen 618.

[0123] Turning to the second side of the transfer adapter, in some embodiments, there may be some structural similarities to the first side, while the second side is configured to couple with a receiving plate. For example, in a transfer system where the receiving plate includes a plurality of receiving holes, the second side of the transfer adapter may include a plurality of secondary extensions configured to engage (e.g., insert into) the receiving holes. In some embodiments, the transfer adapter of the present disclosure includes secondary extensions that protrude from the second surface of the transfer adapter base. For example, Fig. 6E An enlarged view of a portion of the second side (616) of the transfer adapter is shown, including the secondary extension 608 protruding from the second planar surface 612 of the planar sheet 602. Fig. 6Fis a cross-sectional view of a secondary extension 608 inserted into and sealed against the inner surface of the receiving hole 642. In this example, each secondary extension can be aligned with and surround a different opening of the transfer adapter base. In some embodiments, the secondary extension includes a secondary lumen that is configured to fluidly connect with the opening of the transfer adapter base and, in turn, to the main lumen of the main extension. For example, the secondary lumen 620 can be fluidly connected to the opening 604 and the main lumen 618 to form a continuous transfer lumen extending between the first side 614 and the second side 616 of the transfer adapter 600. In some embodiments, the secondary extension can taper from an inner end adjacent to the base (e.g., a flat sheet) to an outer end opposite and away from the base. For example, as Fig. 6F As shown, the secondary extension 608 can taper from an inner end 644 adjacent to the planar sheet 602 to an outer end 646 distal to the planar sheet. In other words, the cross-sectional area of ​​the secondary extension at the inner end can be greater than the cross-sectional area at the outer end.

[0124] In some embodiments, the outer end is configured so that fluid exiting from the outer end into the receiving hole is directed to the wall of the receiving hole rather than to the center of the receiving hole. On the one hand, this configuration reduces or eliminates the effect of the outflowing fluid on one or more contents of the receiving hole. For example, Figure 6G As shown in , the outer end 646 is configured to point toward the wall of the receiving well rather than the center of the receiving well, so that fluid flowing out of the outer end is directed to the wall rather than to the center of the receiving well, which may contain one or more reagents 634. Such an embodiment can reduce the amount of disturbance of the contents at the base of the receiving well 642 caused by fluid transfer. It should be appreciated that the size of the opening of the outer end 646 and its distance from the base of the receiving well 642 can also be varied to eliminate, reduce, or achieve minimal disturbance of the target agent 634 at the base of the bottom of the receiving well. In an example where a cell or tissue culture is grown at the bottom of the receiving well, such as Figure 6G The configuration shown can be used to avoid disturbance of cell or tissue cultures during liquid transfer.

[0125] In other embodiments, one or more reagents may be disposed on the inner wall of the receiving hole rather than on the bottom base of the receiving hole. Depending on the purpose of the fluid transfer, an appropriate secondary extension configuration may be selected. For example, if the transferred fluid is intended to contact one or more reagents on the inner wall (e.g., lyophilized reagents on the reconstitution wall), a secondary extension such as Figure 6G However, if contact and / or interference with the reagents should be avoided (e.g., because cells grow on the walls), the secondary extensions can be configured to direct the fluid toward the bottom plate rather than the inner walls of the receiving well.

[0126] In some embodiments, the tapered shape of the secondary extension allows the outer end to be inserted into the entrance of the receiving hole, so that the opening of each secondary lumen and transfer adapter is aligned with a different receiving hole. Fig. 6F As shown, the secondary extension 608 can be moved forward into the receiving hole 642 until at least a portion of the outer surface of the secondary extension contacts and seals against a portion of the inner surface of the receiving hole. The portion of the secondary extension 608 and the receiving hole 642 that forms a seal (e.g., a fluid-tight seal) can have the same cross-sectional area and shape to form a continuous seal around the perimeter of the secondary extension and the receiving hole. The seal can prevent fluid flow between the receiving holes 642. Although Fig. 6E and 6F The secondary extension 608 shown in FIG. 6 includes only one slope and no slits, but it should be appreciated that the secondary extension may include any of the features described herein with reference to the primary extension, including two or more portions having different slopes, and one or more slits.

[0127] In another aspect, a transfer adapter disclosed herein includes a planar sheet having a first planar surface and a second planar surface, and a plurality of openings extending between the first planar surface and the second planar surface. Fig. 7A As shown, the transfer adapter 700 is positioned above the multiwell plate and a close-up view of a portion of the transfer adapter is shown in FIG. Figure 7B In these examples, the transfer adapter 700 includes a planar sheet 702 having a first planar surface 704 and a second planar surface 706, and a plurality of openings 708 extending between the first planar surface and the second planar surface. In some embodiments, the transfer adapter does not have a reference Figures 6A-6F The primary extension and / or secondary extension described in the transfer adapter embodiment shown in . For example, the transfer adapter 700 may include an opening 708, and the opening 708 is arranged so that each opening is aligned with a different hole of a supply plate 710 (e.g., a 96-well plate). In some embodiments, at least one or all holes of the supply plate may be sealed against the first flat surface of the transfer adapter, which may reduce the chance of fluid mixing between the holes. Therefore, on the one hand, each opening 708 of the transfer adapter 700 may be aligned with a different supply hole of the supply plate, and each supply hole may be sealed against the first flat surface 704 of the transfer adapter. In some embodiments, the first flat surface of the transfer adapter is fixedly connected to the bottom surface of the supply plate (e.g., 300) and / or the bottom surface of the separation hole structure (e.g., 400). The transfer adapter and the supply plate may be formed integrally, optionally including the separation hole structure. Similarly, in an embodiment of a transfer system including a receiving plate having a plurality of receiving holes, each opening (e.g., 708) of the transfer adapter may be aligned with a different receiving hole, and each receiving hole may be sealed against the second flat surface (e.g., 706) of the transfer adapter.

[0128] In one aspect, the transfer adapter can include one or more features to facilitate alignment of the opening with the supply hole of the supply plate and / or the receiving hole of the receiving plate. For example, one or more sides of the transfer adapter 700 can have the same dimensions (e.g., length and / or width) as corresponding sides of the supply plate and / or the receiving plate. The transfer adapter and the supply plate and / or the receiving plate can be configured such that aligning the corresponding sides can also align the opening (e.g., 708) with the supply hole of the supply plate and / or the receiving hole of the receiving plate.

[0129] On the other hand, the transfer adapter (e.g., 700) is configured to enable the supply hole of the supply plate (e.g., 710) to be sealed against the first flat surface (e.g., 704), and / or to enable the receiving hole of the receiving plate to be sealed against the second flat surface (e.g., 706). For example, the first flat surface may be at least partially covered with an adhesive. When the transfer assembly is formed, the first flat surface of the transfer adapter may engage the top surface (e.g., 712) of the supply plate, and the adhesive may reversibly or irreversibly attach the two surfaces and form a liquid-tight seal between them. Therefore, the fluid may be allowed to flow out of the hole of the supply plate through the opening (e.g., 708), but the seal may prevent the fluid from flowing between the first flat surface of the transfer adapter and the top surface of the supply plate to reach different holes. Similarly, the adhesive may at least partially cover the second flat surface of the transfer adapter so as to form a seal with the surface of the receiving plate and prevent the flow of fluid between the receiving holes.

[0130] In some embodiments, the adhesive may cover the entire first planar surface and / or the second planar surface (e.g., 704 and 706, respectively), while in other embodiments, it may be desirable for the adhesive to cover only a portion of the first planar surface and / or the second planar surface. For example, to reduce the risk of adhesive contaminating the contents of the holes of the supply plate (e.g., 710) and / or the receiving holes of the receiving plate, the first planar surface and / or the second planar surface of the transfer adapter may be free of adhesive in areas that may contact the fluid (e.g., around opening 708). It should be appreciated that while the adhesive may cover one or more surfaces of the transfer adapter, the same or different adhesive may additionally or alternatively cover surfaces of the supply plate and / or the receiving plate.

[0131] A transfer adapter (e.g., 700) may be configured to control the amount of force that can cause a fluid to flow through an opening (e.g., 708). For example, the transfer adapter may be configured such that if the transfer assembly is positioned such that a supply plate (e.g., 710) is above the transfer adapter, gravity may not be sufficient to produce a substantial flow of fluid through the opening. In order to cause the fluid to flow through the opening and be transferred from the holes of the supply plate, an external force, such as from a centrifuge, may be applied to the transfer assembly. The size and / or shape (e.g., cross-sectional area and / or width) of at least each opening (708), one or more materials of the transfer adapter, and / or properties of the fluid (e.g., viscosity) may determine the force required to cause the fluid to flow through the opening. For example, reducing the size of the opening, increasing the hydrophobicity of the material interfacing with the opening, and / or increasing the cohesive forces (e.g., surface tension) of the fluid may increase the magnitude of the force required to cause a substantial flow of fluid through the opening. Although Figure 7A-7B The opening 708 shown in FIG. 7 is cross-shaped, but the opening can have any shape (eg, square, rectangular, circular, or oval), and the transfer adapter can still be configured to prevent fluid from flowing through the opening due to gravity. Figure 7C An example of using centrifugal force to transfer fluid into a hole is shown. In one aspect, a pre-cut hole or slit in a transfer adapter (e.g., made of rubber) will seal under normal gravity conditions (e.g., Figure 7C , left), while during centrifugation the pre-cut holes or slits in the rubber open as the increased centrifugal force forces the fluid through the holes or slits (e.g. Figure 7C ,right).

[0132] although Figure 7C One opening is shown for illustration purposes, but in some embodiments, the transfer adapter includes multiple openings, such as Figure 7A-7B On the one hand, multiple subdivisions of the liquid help reduce interference from unequal meniscus structures (where the liquid meets the container wall) in the subdivisions. For example, Fig.7D As shown, without subdivided regions, a larger meniscus structure (and therefore a larger volume) can be seen closest to the wall, while a smaller volume is seen in the center. By providing multiple subdivided regions, equal volumes in each subdivided region can be achieved, and interference from unequal meniscus structures can be reduced or eliminated.

[0133] In some embodiments, the transfer adapter can be configured to have one opening aligned with each hole of the supply plate, while in other embodiments, the transfer adapter can be configured to have more than one opening aligned with each hole of the supply plate. For example, the transfer adapter can be configured so that multiple small openings are aligned with each hole.

[0134] In some embodiments, an external force, such as from a centrifuge, can change the size and / or shape of each opening (e.g., 708), which can increase the flow of fluid through the opening. For example, one or more leaflets (e.g., four leaflets 714) can surround and / or define the area of ​​each opening, and the leaflets can move to change the area of ​​the opening. In the absence of any external force applied to the transfer adapter (e.g., 700), the leaflets can be in a position such as Figure 7A-7B . When the leaflets are in the closed position, the area of ​​each opening may be minimized. The transfer adapter may be configured so that when the area of ​​each opening is minimized and only gravity is applied to the fluid, the fluid may not flow through the opening. In these embodiments, the area of ​​each opening when minimized may be approximately zero (e.g., the opening may be completely closed), or the opening may be configured so that cohesive forces within the fluid and / or adhesion forces between the fluid and the transfer adapter may prevent the fluid from flowing through the opening. When an external force is applied to the transfer adapter, at least a portion of each leaflet may be deflected or otherwise moved out of the horizontal plane of the flat sheet (e.g., 702), and the leaflets may be in a second open position. When the leaflets are in the open position, the area of ​​each opening may be greater than the area when the leaflets are in the closed position, thereby enabling fluid to flow through the opening. The transfer adapter may be configured so that a desired force is required to move the leaflets from the closed position to the open position and allow fluid to flow through the opening. For example, the size, shape, and material of the leaflets may at least partially determine how easily they move.

[0135] It should be appreciated that any of the transfer adapter features described may be combined in any suitable manner. For example, an embodiment of a transfer adapter may include a transfer adapter from reference Figures 6A-6F Some components of the embodiments discussed and from reference Figure 7A-7B Specifically, an embodiment of the transfer adapter may include a primary extension and a secondary extension to facilitate alignment and sealing of the transfer adapter with the supply plate and the receiving plate, respectively. This embodiment may also have an opening, such as Figure 7A-7B The cross-shaped opening 708 shown in the figure is configured to prevent fluid flow unless an external force is applied to the transfer adapter. In addition, the transfer adapter may or may not have an adhesive on one or more surfaces to facilitate sealing and / or coupling of the transfer adapter with the porous plate and / or receiving plate.

[0136] The transfer adapter may include one or more additional or alternative features to facilitate interaction with the supply plate and / or receiving plate. For example, the transfer adapter may include permanent or removable clips or fixtures to assist in coupling, aligning, and / or sealing the transfer adapter to the supply plate and / or receiving plate. In some embodiments, the transfer adapter may be configured to be used with one size or type of supply plate and / or receiving plate. For example, the transfer adapter may have a fixed number of openings, and the number of openings may be the same as the number of openings in one type of supply plate. In other embodiments, one transfer adapter may be used with multiple sizes or types of supply plates and / or receiving plates. For example, the transfer adapter may include one or more parts that can be permanently or reversibly separated and / or connected to reduce or increase the size of the transfer adapter, thereby reducing or increasing the number of openings.

[0137] In any of the foregoing embodiments, the transfer adapter may comprise a suitable material such as, but not limited to, rubber, plastic, polymer, or the like.

[0138] Receiver Board

[0139] In some embodiments, the transfer system described herein may include a receiving plate that can be coupled to a transfer adapter and contain fluid transferred from the holes of a supply plate. When the transfer assembly is formed, the receiving plate can be reversibly attached to the side of the transfer adapter opposite to the side to which the supply plate is attached. Then, the fluid can flow out of the hole of the supply plate, through the opening in the transfer adapter, and into the receiving plate. Including the transferred fluid in the receiving plate can prevent the fluid from escaping into the centrifuge, and can allow the fluid to be studied after it is separated from the target agent. In some embodiments, the receiving plate can be configured to prevent the fluids transferred from different holes of the supply plate from mixing. For example, the receiving plate can include multiple receiving holes, and the fluids transferred from each hole of the supply plate can flow into different receiving holes. In other embodiments, the fluids transferred from different holes of the supply plate can be mixed in the receiving plate. For example, the receiving plate can include only one receiving hole, and the fluids from each hole of the supply plate can flow into the receiving hole.

[0140] The content of the receiving hole of the receiving plate used in the transfer system described herein may include one or more target agents. The target agent may be at least temporarily fixed on or fixed on the inner surface or base of the receiving hole. Through the transfer process, the fluid can be basically transferred from the supply hole to the receiving hole, and the target agent can remain fixed on the inner surface of the receiving hole to a large extent. Any suitable target agent can be used, such as protein, nucleic acid, microorganism (e.g., bacteria, fungi), plant (e.g., algae), virus, small molecule drug or any compound, polymer, antigen, antibody, cell fragment, cell homogenate, DNA, peptide, etc.

[0141] In an embodiment of a receiving plate configured to prevent mixing of fluids transferred from different holes of a supply plate, the number and arrangement of the receiving holes can be the same as the number and arrangement of the openings in the transfer adapter and the holes in the supply plate. Thus, alignment of the receiving plate with the transfer adapter and the supply plate can align each receiving hole with a different opening of the transfer adapter and a different hole of the supply plate. Each receiving hole can have any size and shape discussed with reference to the holes of the supply plate, as long as the volume of each receiving hole can be large enough to accommodate the fluid transferred from one hole.

[0142] To prevent fluid flow between different receiving holes, the receiving plate can be configured to seal against the transfer adapter. For example, in an embodiment of a transfer system including a transfer adapter having a secondary extension, such as Fig. 6A , 6B , 6E and 6F, each secondary extension can be assembled into a different receiving hole and sealed against a different receiving hole. Fig. 6F Described in more detail, at least a portion of the receiving hole may have the same cross-sectional size and shape as at least a portion of the secondary extension, which may facilitate forming a continuous circumferential seal between the outer surface of the secondary extension and the inner surface of the receiving hole.

[0143] In some embodiments of a receiving plate comprising a plurality of receiving holes, the receiving plate may be a standard off-the-shelf multi-well plate. For example, in some embodiments of the transfer system, the receiving plate may be the same as the supply plate. In other embodiments, the receiving plate may be a standard multi-well plate of a different type than the supply plate. For example, the supply plate may be coated with a material to immobilize the target agent, while the receiving plate may not include such a coating, or vice versa.

[0144] Figure 1A and 1B The embodiment of the receiving plate shown in the transfer system in FIG. 1 can be a standard multi-well plate. As seen therein, the receiving plate (108) can include a contact surface (126) and a plurality of receiving holes (118). When the transfer assembly (102) is formed, the contact surface can be configured to face or contact the transfer adapter (106). Fluid can flow into each receiving hole (118) through an opening or inlet (124).

[0145] In some embodiments, the receiving plate may allow mixing of fluids transferred from different holes of the supply plate. For example, the receiving plate may include fewer receiving holes than the openings in the transfer adapter and the holes in the supply plate, and / or a plurality of receiving holes may be fluidically connected. A receiving plate configured to allow mixing of fluids transferred from only some holes of the supply plate may allow mixing between fluids transferred (for example, half of the holes may flow into the first receiving hole, and the other half of the holes may flow into the second receiving hole, each row or column of holes may flow into different receiving holes, etc.). In other embodiments, the receiving plate may be configured to allow mixing of fluids transferred from each hole of the supply plate. For example, the receiving plate may include a single receiving hole, and the fluid transferred from each hole of the supply plate may flow into a single receiving hole. In other embodiments, the receiving plate may be adjustable to change the number of receiving holes, which may change the degree of fluid mixing allowed. For example, the receiving plate may include one or more removable, combinable and / or separable walls, which may be adjusted to change the number of receiving holes.

[0146] On the other hand, the receiving plate may include one or more features to facilitate alignment, sealing and / or coupling with the transfer adapter. For example, even in the embodiment of the receiving plate that allows the fluids transferred from the different holes of the supply plate to mix, it may be advantageous for the receiving plate to seal against the transfer adapter to prevent fluid from leaking (e.g., leaking into the centrifuge) from the transfer system. In certain embodiments, at least a portion of the contact surface of the transfer adapter may be covered with an adhesive to facilitate sealing. The adhesive may maintain the contact between at least a portion of the contact surface of the receiving plate and the surface of the transfer adapter, and prevent fluid from moving between the two surfaces. Additionally or alternatively, a clamp or fixture may keep the contact surface of the receiving plate and the surface of the supply plate together to prevent fluid from flowing between the receiving holes. In certain embodiments, the receiving plate may have any male or female structure that interacts with the corresponding female or male structure on the transfer adapter, respectively, to facilitate alignment, sealing and / or coupling of the plate. For example, the receiving plate may include a lip around the periphery of the contact surface, around one or more receiving holes or at any other suitable position, and the lip may be assembled into the corresponding groove of the transfer adapter.

[0147] In certain embodiments, except being connected with the transfer adapter, the receiving plate can be configured to be removably connected with the supply plate.For example, it may be advantageous that the receiving plate can also be used as the cover of the supply plate.In other words, the receiving plate can be configured to be reversibly attached to the supply plate so that the top surface of the supply plate is covered.In certain embodiments, the receiving plate can be configured to be placed in a centrifuge.For example, the shape and size of the receiving plate can be set to be assembled into the centrifuge (for example, assembled into the bucket of the centrifuge) and / or include features (for example, clips or fixtures) to be reversibly attached to a part of the centrifuge (for example, the bucket of the centrifuge).The receiving plate can include any suitable one or more materials, including but not limited to glass, plastics and metal.

[0148] Although the receiving plate is generally described as a separate and distinct element from the transfer adapter, it should be appreciated that in some embodiments, the receiving plate can be integrally formed with the transfer adapter. In these embodiments, the combined transfer adapter-receiving plate can be coupled to the supply plate to facilitate the transfer of fluid from the holes of the supply plate and to contain the transferred fluid. The combined transfer adapter-receiving plate can be configured to allow or prevent mixing of fluids transferred from different holes of the supply plate.

[0149] Transfer Systems and Kits

[0150] One or more elements of the transfer system described herein may be included in a transfer kit, which may facilitate simultaneous and consistent transfer of fluid from the holes of a supply plate to the holes of a receiving plate. For example, in some embodiments, the transfer kit may include a supply plate, a transfer adapter, and a receiving plate. In other embodiments, the transfer kit may include a transfer adapter and a receiving plate, and the transfer kit may be configured to be used with a standard ready-made porous plate. In an embodiment where a transfer adapter and a supply plate are integrally formed, the transfer kit may include a combined supply plate-transfer adapter. In an embodiment where a transfer adapter and a receiving plate are integrally formed, the transfer kit may include a combined transfer adapter-receiving plate. In some embodiments, the transfer kit may further include a separation hole structure configured to be coupled to the supply plate. The separation hole structure may be configured to be removably coupled to the supply plate. In some embodiments, the separation hole structure is fixedly attached or integrated with the supply plate. In some embodiments, the transfer kit may include a transfer adapter, and the transfer kit may be configured to be used with a standard porous plate for both a supply plate and a receiving plate. It should be appreciated that the transfer kit may be configured to be used with a specific target agent, fluid, centrifuge, and / or a standard porous plate type.

[0151] In some embodiments, the reagent transfer system disclosed herein includes a supply container for well-to-well transfer into a receiving container. In one aspect, the supply container is a supply plate that includes a structure used as a transfer adapter as disclosed in any of the aforementioned embodiments. For example, in some embodiments, the transfer adapter structure of the supply plate is similar to Figure 6B The secondary extension (608) shown; however, in these embodiments, as Figure 6A-6B The main extension (606) is shown now having a closed end - the transfer adapter is thus configured to receive one or more reagents and act as a supply plate. Fig. 9As shown, one or more reagents (not shown) can be pre-deposited (e.g., by a manufacturer) into the supply plate (900). In one aspect, the supply plate faces upward when the one or more pre-deposited reagents are positioned at the bottom of one or more closed ends (e.g., wells). In another aspect, the supply plate is configured to couple with the receiving plate (902) to form a transfer assembly (904). These examples use a 96-well plate format for illustrative purposes, but any well format (e.g., a 384-well plate format) can be used. In one aspect, the supply plate can be inverted with the receiving plate facing upward to form a transfer assembly, such as Fig. 9 As shown. On the other hand, the receiving plate can be inverted with the supply plate facing upward to form a transfer assembly. In other words, the supply plate can be mounted on top of the receiving plate, or vice versa, for example, by inserting the transfer adapter structure of the supply plate into one or more wells of a receiving container (e.g., a 96-well plate). This step can also be performed by the manufacturer.

[0152] In one aspect, the transfer assembly can be subjected to an external force, such as a centrifugal force, to transfer one or more reagents from a supply container to a receiving container. Fig. 10A An example is shown in FIG, where reagents 1004 are pre-deposited in a supply plate 1000, which is assembled onto a receiving plate 1002. Once the assembly is placed in a centrifuge, the reagents 1004 can be transferred to the wells of the receiving plate 1002.

[0153] On the one hand, a device for transferring fluid is provided herein, comprising: a flat sheet having a first flat surface on a first side and a second flat surface on a second side, a plurality of compartments (e.g., holes) on the first side having an opening on the second side, and a plurality of extensions on the second side protruding from the second flat surface and including an inner cavity connected to the opening of the compartment. In one embodiment, each extension is configured to be inserted into a different receiving hole of a receiving plate. In another embodiment, the outer surface of each extension is configured to seal against a different receiving hole of a receiving plate. In any of the foregoing embodiments, the inner surface of the extension may be configured to form an angle with the inner wall of the receiving hole, and the angle may be less than about 10 degrees, for example, about 1 degree, about 2 degrees, about 3 degrees, about 4 degrees, about 5 degrees, about 6 degrees, about 7 degrees, about 8 degrees, about 9 degrees, or about 10 degrees.

[0154] On the other hand, the supply plate includes one or more structures or patterns to better hold and / or distribute reagents.For example, projections may be provided in the holes of the supply plate (e.g., at the bottom of the holes) to accommodate reagents, such as dried reagents, such as freeze-dried reagents. Fig. 10BAn example is shown in Figure 1, wherein reagent 1004 is pre-deposited on protrusion 1006 in supply plate 1000, and supply plate is assembled on receiving plate 1002. Once assembly is placed in centrifuge, reagent 1004 can be transferred to the hole of receiving plate 1002. In this example, before (for example, by centrifugation) transfer, protrusion 1006 can facilitate reagent 1004 to be kept in supply plate. During transfer, protrusion 1006 can help reagent 1004 to be distributed in receiving plate. Protrusion of any suitable pattern etc. can be used. For example, supply plate can include multiple protrusions in one or more holes, each with different reagents pre-deposited thereon. Receiving plate can include one or more solutions in receiving hole. Once transfer assembly is formed, material transfer to receiving hole and / or from receiving hole can be performed. For example, when leakproof seal is formed between supply hole and receiving hole, solution in receiving hole can be mixed with one or more reagents on multiple protrusions of supply hole.

[0155] In some embodiments, the kit includes a system for delivering materials or reagents to implement the methods disclosed herein. In the context of a reaction experiment using a currently disclosed material transfer system, the kit may include a system that allows storage, transportation or delivery of reaction reagents (e.g., samples, enzymes, etc. in appropriate containers) and / or support materials (e.g., buffer, written instructions for performing assays, etc.) from one location to another. For example, the kit includes one or more compartments (e.g., boxes) containing relevant reaction reagents and / or support materials. These contents may be delivered to the intended recipient together or individually. For example, a first container may include an enzyme for assaying, and a second container includes a sample. In some embodiments, each component of the kit (e.g., a supply plate, a transfer adapter, and a receiving plate) may be packaged separately. In other embodiments, two or more components of the kit may be packaged together. For example, a supply plate, a transfer adapter, and / or a receiving plate may be packaged together. Alternatively, a supply plate and a transfer adapter may be packaged together, and a receiving plate may be packaged separately or provided by the user of the kit.

[0156] method

[0157] In one aspect, disclosed herein is a method for facilitating the transfer of materials (e.g., one or more fluids, and / or one or more dry reagents) to and / or from the wells of a multiwell plate using fluid transfer forces generated, for example, by gravity or a centrifuge.

[0158] In some embodiments, it is desired to remove fluid from a plurality of wells of a supply plate, and the method includes transferring the fluid from the wells of the supply plate to a receiving plate.

[0159] In some embodiments, it is desired to add the fluid to a plurality of wells in a receiving plate, and the method includes first adding the fluid to a donor plate and then transferring the fluid from the donor plate to the wells of the receiving plate.

[0160] In any of the foregoing embodiments, the supply plate may have a feature that allows fluid to be loaded into multiple holes in the supply plate at the same time. Fluid can be transferred from each hole of the supply plate at the same time, and the influence of the fluid transfer force can be basically consistent within each hole and between different holes. In addition, the fluid transfer may be mainly automatic and may require minimal manual work. The method may generally include forming a transfer assembly by a supply plate, a transfer adapter and a receiving plate, and a centrifugal transfer assembly. Forming the transfer assembly may include removably connecting the first side of the transfer adapter to the supply plate, and removably connecting the opposite second side of the transfer adapter to the receiving plate. Connecting the transfer adapter to the supply plate may involve aligning each opening of the transfer adapter with different holes of the supply plate and forming a liquid-tight seal between the transfer adapter and the supply plate. Similarly, in an embodiment of a transfer system including a receiving plate with multiple receiving holes, connecting the transfer adapter and the receiving plate may involve aligning each opening of the transfer adapter with different receiving holes of the receiving plate and forming a liquid-tight seal between the transfer adapter and the receiving plate. In some embodiments, the supply plate and the transfer adapter are integrally formed, and forming the transfer assembly may include removably coupling an exposed side of the transfer adapter of the combined supply plate-transfer adapter to the receiving plate. In some embodiments, the transfer adapter and the supply plate are integrally formed, and forming the transfer assembly may include removably coupling an exposed side of the transfer adapter of the combined transfer adapter-receiving plate to the supply plate.

[0161] The transfer assembly can be in a first position when it is formed. In the first position, the transfer adapter can be positioned below the receiving plate and above the supply plate. In this position, the fluid may remain in the hole of the supply plate. In certain embodiments, such as the transfer assembly of the supply plate 300 including Fig. 3, in the first position, the transfer adapter can be positioned below the supply plate and above the receiving plate, and in this position, the fluid can remain in the hole of the supply plate. After the transfer assembly is formed, it can be centrifuged around the axis of rotation. The centrifuge can generate a fluid transfer force, which can cause the fluid to move away from the axis of rotation. Therefore, the transfer assembly can be oriented in the centrifuge so that when the centrifuge spins, the outward movement of the fluid can cause the fluid to flow from the supply plate to the receiving plate. This can be achieved by orienting the transfer assembly so that the supply plate is closer to the axis of rotation than the transfer adapter and the receiving plate. When the centrifuge is stationary and when it spins, the specific orientation of the transfer assembly (for example, the angle of the transfer assembly relative to the axis of rotation) can at least depend on the type of the centrifuge rotor used. The transfer assembly is centrifuged to control the amount and / or rate of fluid transfer within the expected duration and at the expected speed. After centrifuging the transfer assembly, the donor plate, transfer adapter, and receiving plate can be separated, which can, for example, allow access to the transferred fluid and / or isolated target agent.

[0162] Forming transfer components

[0163] In some embodiments of the methods described herein, the supply plate does not contain fluid, and the method includes delivering the fluid to the supply plate. With reference to the embodiment of the supply plate 300, the fluid (e.g., aqueous composition) can be delivered to the holding chamber (e.g., the holding chamber 350) of the supply plate. The holding chamber can be formed by connecting the boundary wall (e.g., the boundary wall 302) to the transfer adapter (e.g., the transfer adapter 700). In some embodiments, the boundary wall is fixedly attached or integrated with the transfer adapter. A separation hole structure (e.g., the separation hole structure 400 described above) can be connected in the holding chamber, which can divide the contents of the holding chamber into a plurality of separation holes (e.g., the separation holes 408 described above). In some embodiments, the separation hole structure is configured to be removably connected in the holding chamber, and the fluid is delivered to the holding chamber before the separation hole structure is connected in the holding chamber. In some embodiments, the fluid is delivered to the holding chamber after the separation hole structure is connected in the holding chamber. In some embodiments, the separation hole structure is fixedly attached or integrated with the boundary wall and / or the transfer adapter. In some embodiments where the fluid is delivered to a holding chamber having a separation hole structure therein, the separation hole structure may include one or more separation wall grooves (e.g., separation wall grooves 440 as described above) such that two or more separation holes are fluidically connected, and delivering the fluid to the holding chamber includes delivering the fluid to the one or more separation holes. In some embodiments, a period of time may be allowed to allow an equal volume of fluid to be distributed to each separation hole that is fluidically connected.

[0164] In some embodiments, the transfer assembly can be formed by coupling at least one receiving plate to a supply plate including a plurality of holes filled with a fluid, optionally wherein the holes of the supply plate and / or the receiving plate contain one or more target agents. For example, the first side of the transfer adapter can be coupled to the supply plate (e.g., removably, fixedly or integrally), and the relative second side of the transfer adapter can be coupled to the receiving plate. On the one hand, the supply plate can be directly coupled to the transfer adapter and indirectly coupled to the receiving plate. The supply plate, the transfer adapter and the receiving plate are coupled to form a transfer assembly, which allows the fluid from the hole of the supply plate to be transferred through the opening in the transfer adapter and enter the receiving plate. The supply plate, the transfer adapter and the receiving plate can be coupled in any order. In other words, the supply plate can be attached to the first side of the transfer adapter before, after or simultaneously when the receiving plate is attached to the second side of the transfer adapter. The coupling or attachment between the transfer adapter and the supply plate and / or the receiving plate can be irreversible or reversible. The reversible coupling of the transfer adapter and the supply plate and / or the receiving plate allows the plates to be separated and separated, so that, for example, the fluid transferred and / or the separated target agent can be taken after the transfer.

[0165] Connecting the transfer adapter and the supply plate may include aligning the transfer adapter and the supply plate and sealing them together. Aligning the transfer adapter and the supply plate may include aligning an opening of the transfer adapter with a hole of the supply plate. This alignment may be facilitated by one or more features of the transfer adapter and / or the supply plate. For example, in an embodiment of a transfer adapter that includes a main extension associated with each opening, each main extension may be inserted into a different hole of the supply plate, thereby aligning each opening with a different hole. In an embodiment of a transfer adapter that includes an alignment guide, the alignment guide may be aligned with a predetermined portion of the supply plate, which may also align the opening of the transfer adapter with the hole of the supply plate.

[0166] The coupling of the transfer adapter and the supply plate may also include forming a fluid-tight seal between the two plates. One or more seals may prevent fluid from flowing between different holes in the supply plate, but may still allow fluid to flow out of the hole and through the opening in the transfer adapter. The seal may be facilitated by one or more features of the transfer adapter and / or the supply plate. In some embodiments, these one or more features may be the same as one or more features that facilitate alignment. For example, in an embodiment of a transfer adapter including a main extension associated with each opening, each main extension may form a seal with a different hole of the supply plate in addition to facilitating alignment. Each main extension may be inserted into the hole and moved forward until a portion of the outer surface of the main extension seals against a portion of the inner surface of the hole. In some embodiments, one or more features that facilitate sealing may be different from one or more features that facilitate alignment. For example, some embodiments of the transfer adapter may include a first surface at least partially covered with an adhesive. In this embodiment, the first surface of the transfer adapter covered with an adhesive may be pressed against the top surface of the supply plate to form a seal that prevents fluid from flowing between different holes. In some embodiments, the transfer adapter is fixedly attached or integral with the supply plate. It should be recognized that in an embodiment in which the transfer adapter and the supply plate are integrally formed, the method of forming the transfer assembly may not include coupling the transfer adapter and the supply plate.

[0167] Forming the transfer assembly may also include coupling the transfer adapter and the receiving plate. Connecting the transfer adapter and the receiving plate may include aligning the transfer adapter and the receiving plate and sealing them together. In an embodiment of a transfer assembly including a receiving plate having a plurality of receiving holes, the method of aligning and / or sealing the transfer adapter and the receiving plate may be similar to the method described for aligning and / or sealing the transfer adapter and the supply plate. For example, aligning the transfer adapter and the receiving plate may include aligning the opening of the transfer adapter with the receiving hole of the receiving plate. Sealing the transfer adapter and the receiving plate together may form a liquid-tight seal between the receiving hole and the transfer adapter, which may allow fluid to flow through the opening and enter the receiving hole, but may prevent fluid from flowing between the receiving holes. These seals may also help ensure that fluid from each hole of the supply plate flows into different receiving holes of the receiving plate. It should be appreciated that in some embodiments, coupling the transfer adapter and the receiving plate may include aligning each opening with a different receiving hole, but may not form a liquid-tight seal. One or more features of the transfer adapter and / or the receiving plate may facilitate the alignment and / or sealing of the transfer adapter and the receiving plate, such as a secondary extension and / or adhesive on the second side of the transfer adapter. In some embodiments, the transfer adapter is fixedly attached or integrated with the receiving plate.

[0168] In embodiments of a transfer assembly that includes a receiving plate having fewer receiving holes than openings in a transfer adapter, the coupling of the transfer adapter and the receiving plate may still include alignment and / or sealing. For example, when a transfer adapter that includes multiple openings is coupled to a receiving plate that includes a single receiving hole, the alignment may align all of the openings with the single receiving hole so that all of the transferred fluid may be contained in the receiving hole. Additionally, a fluid-tight seal between the transfer adapter and the receiving plate may be formed around the single receiving hole to prevent fluid from escaping from the transfer assembly. It should also be appreciated that in embodiments in which the transfer adapter and the receiving plate are integrally formed, the method of forming the transfer assembly may not include coupling the transfer adapter and the receiving plate.

[0169] In some embodiments, the transfer assembly may be Figure 1B A first upright position assembly is shown, wherein the feed plate is in an upright position, e.g., wherein the openings of the apertures face upward. Figure 1B An example of a transfer assembly (102) in a first upright position is shown. As shown, the transfer adapter 106 is positioned below the receiving plate 108 and above the supply plate 104. The opening of the hole (not shown) of the supply plate faces upward and toward the transfer adapter. The receiving plate is inverted, for example, the entrance of the receiving hole faces downward toward the transfer adapter. In this position, the fluid can be retained in the hole of the supply plate due to gravity.

[0170] See also Figure 1DIn the embodiment of the transfer assembly (152) shown in , the transfer assembly can be assembled in a first position, wherein the transfer adapter 156 is positioned below the supply plate 154 and above the receiving plate 158. The supply plate is in an upright position, for example, wherein the top opening of the hole faces upward, and the bottom opening of the hole (not shown) faces downward and toward the transfer adapter. The receiving plate is upright, for example, the entrance of the receiving hole faces upward toward the transfer adapter. In this position, due to the characteristics of the transfer adapter and / or the fluid, the fluid can be retained in the hole of the supply plate. In some embodiments, the supply plate is fixedly attached or integrated with the transfer adapter. In some embodiments, the supply plate is a supply plate (e.g., 300) and includes a separation hole structure (e.g., 400) coupled thereto. In some embodiments, the separation hole structure is configured to be removably coupled to the supply plate, and assembling the transfer assembly further includes coupling the separation hole structure to the supply plate. In some embodiments, the separation hole structure is fixedly attached or integrated with the supply plate.

[0171] Material Transfer

[0172] After the transfer assembly has been formed in the first position, the transfer assembly can be repositioned to allow gravity and / or external force to transfer fluid from the supply plate to the receiving plate. For example, in some embodiments of the method described herein, the transfer assembly can be placed in a centrifuge, and the centrifuge can generate a fluid transfer force. The fluid transfer force can have a substantially simultaneous and consistent impact on the fluid and any target agent in each hole and in different holes. In the embodiment of the transfer assembly in which fluid does not flow from the supply plate to the receiving plate under gravity, the centrifugal treatment transfer assembly can cause fluid to flow from the supply plate to the receiving plate. In the embodiment of the transfer assembly in which fluid flows from the supply plate to the receiving plate under gravity, the centrifugal treatment transfer assembly can cause fluid to flow faster and / or can allow fluid to transfer more completely from the supply plate.

[0173] The centrifuge may include a rotor that spins around an axis of rotation, and a transfer assembly may be placed in or on the rotor to also spin around the axis of rotation. The spinning motion of the transfer assembly may cause the fluid to move outward away from the axis of rotation. The transfer assembly may be oriented in a way in the centrifuge so that the outward movement of the fluid may cause the hole of the supply plate to flow out, through the opening of the transfer adapter, and flow into the receiving plate. For example, when the centrifuge spins, for example, when the centrifuge rotor and the transfer assembly spin around the axis of rotation, the transfer assembly may be oriented so that the supply plate is positioned closer to the axis of rotation than the transfer adapter and the receiving plate. More specifically, when the centrifuge spins, each hole of the supply plate may be positioned closer to the axis of rotation than the opening and the receiving hole of the transfer adapter aligned therewith.

[0174] In some embodiments, Fig. 8AAs shown, when the centrifuge is spinning, the transfer assembly (800) can be oriented parallel to the axis of rotation (802). Figure 8B As shown, when the centrifuge is spinning, the transfer assembly can be oriented at angles between parallel and perpendicular to the axis of rotation. Fig. 8A and 8B As shown, when the centrifuge is spinning, the supply plate (804) may be closer to the axis of rotation than the transfer adapter (806) and the receiving plate (808).

[0175] The specific orientation of the transfer assembly when it is in the centrifuge can depend on the type of centrifuge rotor being used. For example, when a fixed angle rotor is used, the transfer assembly can have the same orientation (e.g., vertical, at an angle between vertical and horizontal) when it is initially placed in the centrifuge and the centrifuge is stationary as when the centrifuge is spinning. In contrast, when a swing bucket rotor is used, the transfer assembly can have a different orientation when it is initially placed in the centrifuge than when the centrifuge is spinning. For example, when the transfer assembly is initially placed in a centrifuge with a swing bucket rotor, the transfer assembly can be oriented horizontally (e.g., perpendicular to the axis of rotation). When the centrifuge is spinning, the barrels of the rotor and the transfer assembly can be tilted outward to orient the transfer assembly so that the feed plate is closer to the axis of rotation than the transfer adapter and receiving plate. It should be appreciated that when the transfer assembly is initially placed horizontally into the centrifuge, the transfer assembly can be inverted compared to its orientation when assembled in the first upright position. For example, as Figure 8C As shown, when the transfer assembly (800) is in the second inverted position, the transfer adapter (806) may be positioned below the supply plate (804) and above the receiving plate (808).

[0176] On the one hand, after the transfer assembly is placed in the centrifuge, the centrifuge rotor can spin to generate a fluid transfer force. The fluid transfer force can cause the fluid to flow outward from the supply plate to the receiving plate, but the target agent can remain attached to the hole of the supply plate and / or the receiving plate. Specific centrifugal method can depend on the characteristics of the transfer assembly (for example, the embodiment of the supply plate, and / or the embodiment of the transfer adapter), the fluid to be transferred (for example, amount and / or viscosity), and any target agent (for example, the attachment of the target agent to the hole, and / or the stability of the target agent). At least based on these characteristics, the transfer assembly can be centrifuged in the desired duration and with the desired centrifuge setting. For example, speed (for example, revolutions per minute), acceleration, temperature, distance from the axis of rotation, the type of rotor and / or any other available settings can be specified to remove fluid from a certain transfer assembly. Settings such as these can be adjusted to increase the chance of substantially all fluids being transferred from the hole of the supply plate, reduce the chance of any target agent being separated or damaged from the hole, and / or determine the time required to remove fluid from the hole.

[0177] As described above, the fluid transfer force generated by the centrifuge may affect different embodiments of the transfer assembly differently. For example, when some embodiments of the transfer assembly are placed in a centrifuge, the fluid transfer force may cause the fluid to begin to flow out of the holes of the supply plate. In other embodiments, the fluid may have begun to flow out of the holes due to gravity before the fluid transfer force from the centrifuge is applied to the transfer assembly. Applying the fluid transfer force to these embodiments of the transfer assembly may cause the fluid flow rate (e.g., volume flow rate and / or flow rate) out of the holes of the supply plate to increase until substantially all of the fluid has been transferred from the supply plate to the receiving plate. When the fluid transfer force is applied to certain embodiments of the transfer assembly, the size and / or shape of the opening of the transfer adapter may change. For example, when the fluid transfer force is applied to a transfer assembly including a transfer adapter having one or more leaflets around each opening, at least a portion of the one or more leaflets may deflect out of the plane of the flat sheet of the transfer adapter and away from the axis of rotation. The movement of the transfer adapter leaflets may increase the size of the opening, thereby allowing increased fluid to flow through the opening. Thus, while the specific effect of the fluid transfer force may depend on the embodiment of the transfer assembly, the overall effect may be to increase the amount or rate of fluid transfer from the holes of the donor plate to the receiver plate.

[0178] When centrifugation transfer assembly, substantially all fluids and any target agent in the transfer assembly can be subjected to the influence of fluid transfer force simultaneously. This is contrasted with other fluid transfer techniques, such as the pipetting of a subset of a single hole or a hole, wherein the fluid transfer force may first affect the fluid and any target agent in some holes before other holes. Simultaneously, all hole transfer fluids from the supply plate may have one or more advantages, such as increasing the speed of fluid transfer and reducing the variability between the reaction, combination or other processes that occur in different holes. For example, when the process is time-sensitive, this may be desirable. In addition, the influence of fluid transfer force can be basically consistent in each hole and between different holes. Therefore, the fluid and any target agent in each hole and in different holes may be subjected to roughly the same fluid transfer force, which may reduce the variability between the samples from different parts of the same hole and between the samples from different holes. By contrast, when using a pipette to remove fluid from a hole, the force near the pipette tip may be different from the force away from the tip, and may be difficult to apply the same force to different holes. It should be appreciated that while the effects of fluid transfer forces generated by the centrifuge may be substantially consistent, there may be negligible embodiments within and between wells due to slight differences in distance from the axis of rotation. However, these differences may have negligible effects on the fluid, target agent, and / or processes occurring in the wells (e.g., reactions, binding, etc.).

[0179] In some embodiments of the methods described herein, the transfer assembly may not be placed in a centrifuge. For example, as described in detail above, some embodiments of the transfer adapter may be configured to allow fluid to flow from the supply plate to the receiving plate under gravity. In one aspect, inverting the transfer assembly from a first upright position to a second inverted position may result in fluid transfer. See Figure 8C , when the transfer assembly (800) is in the second inverted position, the transfer adapter (806) is below the supply plate (804) and above the receiving plate (802). In this orientation, the supply plate can be in an inverted position so that the opening of the hole (not shown) faces downward and toward the transfer adapter. The receiving plate (808) can be in an upright position so that the receiving hole entrance (not shown) faces upward toward the transfer adapter. The transfer assembly can remain in the second inverted position for a desired duration, which may be sufficient to remove substantially all of the fluid from the hole of the supply plate. The desired duration may be determined at least by the characteristics of the transfer assembly (e.g., an embodiment of the supply plate, and / or an embodiment of the transfer adapter), the fluid to be transferred (e.g., the amount and / or viscosity), and any target agent (e.g., the attachment of the target agent to the hole, and / or the stability of the target agent). In addition, it should be recognized that in other embodiments of the methods described herein, the transfer assembly can be flipped from the first upright position to the second inverted position, which may result in fluid transfer, and the transfer assembly can then be subsequently centrifuged as described above.

[0180] Separate transfer components

[0181] In the embodiment of the centrifugal transfer assembly, after centrifugal treatment, the transfer assembly can be removed from the centrifuge, and one or two of the detachable plates. In the embodiment that does not include the method for centrifugal treatment, after the transfer assembly has been in the second inverted position within the desired duration, one or two of the plates of the detachable transfer assembly. In certain embodiments, the detachable supply plate, transfer adapter and receiving plate. The supply plate, transfer adapter and receiving plate can be separated from each other in any order, and this separation can allow the use of the target agent in the supply plate and / or the transfer fluid in the receiving plate. The target agent and / or the fluid of the transfer of isolation can be studied then, or the target agent or the fluid of the transfer can be discarded. In certain embodiments, if for example determine that some fluids remain in the hole of the supply plate, the transfer assembly can be reformed and inverted and / or centrifuged again. In other embodiments, the supply plate can be separated from the transfer adapter, and the transfer adapter and the receiving plate remain connected. In some other embodiments, the receiving plate can be separated from the transfer adapter, and the supply plate and the transfer adapter remain connected.

[0182] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it is apparent that certain changes and modifications are possible and are intended to fall within the scope of the appended claims. In addition, it should be understood that the components and features of the devices described herein may be used in any combination, and the description of certain elements or features with reference to a particular drawing is not intended to limit or imply that the element may not be used in combination with any other described element.

[0183] Examples

[0184] Example #1

[0185] As an example, the methods described herein can be used to study the pathways of drugs.

[0186] 1. Cell introduction: The cell suspension can be pipetted into the wells of a 96-well multiwell plate filled with cell culture medium. The cells can then be allowed to attach to the bottom surface of the wells for a period of time.

[0187] 2. Collection of conditioned medium: After the cells have reached the desired condition, a transfer assembly can be formed by coupling the first surface of a transfer adapter (such as a transfer adapter with 96 openings) to the top of a 96-well multiwell plate (here, a supply plate) in an upright position, and coupling an inverted receiving plate with one hole to the second surface of the transfer adapter so that the opening of the transfer adapter is aligned with the hole of the 96-well multiwell plate. The transfer assembly can be inverted, loaded into a swing bucket rotor, and centrifuged using conditions that allow approximately half of the conditioned medium to be transferred from the supply plate to the receiving plate without significantly disturbing the attached cells. The receiving plate can be separated from the transfer assembly and the conditioned medium is retained. The remaining transfer adapter-96-well multiwell plate assembly can be inverted to make the 96-well multiwell plate upright, and optionally centrifuged using parameters that allow any remaining medium to be collected to the bottom of the well without disturbing the attached cells, and the transfer adapter can be separated from the 96-well multiwell plate.

[0188] 3. Processing: Test conditions can be applied to cells. For example, a combined supply plate-transfer adapter having openings on the top and bottom surfaces of a supply plate can be connected to a 96-well multi-well plate (here, a receiving plate) containing cells to be treated by connecting the exposed transfer adapter surface of the supply plate-transfer adapter to the top of the 96-well multi-well plate in an upright position, thereby forming a transfer assembly. On the one hand, the holding cavity of the supply plate does not have a separation hole structure connected therein before being loaded with fluid. A test agent (such as a drug of a desired concentration) can be added to the retained conditioned medium, which is loaded into the holding cavity of the supply plate. Then, the separation hole structure can be connected in the holding cavity so that the fluid in the holding cavity is divided into 96 separation holes, which are aligned with the openings in the transfer adapter and the holes of the 96-well multi-well plate. The connection of the separation hole structure in the holding cavity can result in a uniform distribution of the fluid volume in each separation hole.

[0189] For example, the separation well structure can be connected in the holding cavity before the test agent diluted in the conditioned medium is loaded. As another example, the separation well structure can include one or more separation wall grooves or gaps that fluidically connect two or more separation wells, and the test agent diluted in the conditioned medium can be loaded into one or more separation wells, allowing the culture medium to be evenly distributed in the fluidically connected separation wells.

[0190] For example, the separation well structure can be configured such that each separation well is fluidly connected to each other, and a composition including conditioned medium supplemented with a test agent can be delivered to one or more separation wells such that the composition is evenly distributed between each separation well.

[0191] For example, the separation well structure can be configured so that a first subset of separation wells (such as the first half) is fluidically connected to each other, and a second subset of separation wells (such as the second half) is fluidically connected to each other, but the first subset of separation wells is not fluidically connected to the second subset of separation wells. In such an example, a first composition comprising a portion of a conditioned medium supplemented with a test agent at one concentration can be delivered to one or more wells of the first subset of separation wells so that the first composition is evenly distributed between each of the first subset of separation wells, and a second composition comprising another portion of a conditioned medium supplemented with a different concentration of a test agent (or different test agents) can be delivered to one or more wells of the second subset of separation wells so that the second composition is evenly distributed between each of the second subset of separation wells.

[0192] The fluid in the supply plate can be transferred to the 96-well multiwell plate containing the cells to be processed by centrifuging the transfer assembly under conditions that allow the fluid to pass from the separation wells of the supply plate through the openings in the transfer adapter and into the wells of the 96-well multiwell plate without substantially disturbing the cells attached thereto.

[0193] 4. Collection of culture medium: After the test conditions are mature, the culture medium from each well can be collected into a separate well of a receiving plate for analysis, such as for the presence of secreted factors. The transfer assembly can be formed as described above in step 2, but wherein the receiving plate has 96 wells that align with the openings of the transfer adapter and the wells of the 96-well multiwell plate containing attached cells. The transfer assembly can be centrifuged under conditions that allow fluid to be transferred from the 96-well multiwell plate to the receiving plate without substantially disturbing the attached cells. The recovered culture medium in the receiving plate can then be analyzed.

[0194] 5. Cell preparation: Cells can be screened for various markers. Cells can be prepared for analysis first. Cells can be washed with phosphate-buffered saline by assembling the transfer assembly described in step 3 above, wherein the phosphate-buffered saline is loaded into the holding chamber of the supply plate, and the transfer assembly is centrifuged under conditions that allow the phosphate-buffered saline to be transferred to the wells of the 96-well multi-well plate without substantially interfering with the attached cells. Then, the transfer assembly described in step 2 above is formed by separating the 96-well multi-well plate from the transfer assembly, and the transfer assembly is centrifuged under conditions that allow the phosphate-buffered saline to be transferred to the receiving plate without substantially interfering with the attached cells, and the phosphate-buffered saline can be removed from the wells of the 96-well multi-well plate. Similar methods of loading fluids into and removing fluids from the wells of the 96-well multi-well plate can be used to fix cells in the 96-well multi-well plate with formalin or paraformaldehyde solutions by loading, incubating, and removing solutions. Then, the cells can be washed with phosphate-buffered saline, blocked with serum or albumin solutions, permeabilized with TritonX-100 if necessary, and then immersed in phosphate-buffered saline using the above method.

[0195] 6. Analyte introduction: One or more analytes (e.g., primary antibodies) may then be delivered using, for example, methods similar to those used to deliver fluids to the wells of a 96-well multiwell plate in step 3 above. Sufficient incubation time may be allowed so that the analyte (e.g., primary antibody) can attach to its target.

[0196] 7. Introduction of detection agent: After washing with phosphate buffered saline using, for example, a method similar to that described above in step 5, the activity of the analytical agent (such as the primary antibody) can be analyzed. For example, if an unconjugated primary antibody is used, the phosphate buffered saline can be replaced with a secondary antibody.

[0197] Example #2

[0198] As another example, the methods described herein can be used to assess the efficacy of a drug on multiple cell types from an individual (eg, a patient).

[0199] 1. Cell introduction: Cell banks from individuals can be pipetted into individual wells of a 96-well multiwell plate. The cells can then be allowed to attach to the bottom surface of the wells for a period of time.

[0200] 2. Drug loading: A supply plate-transfer adapter combination having openings on the top and bottom surfaces of the supply plate can be connected to a 96-well multiwell plate (here, a receiving plate) containing cells to be treated by connecting the exposed transfer adapter surface of the supply plate-transfer adapter to the top of the 96-well multiwell plate in an upright position, thereby forming a transfer assembly. In some cases, the holding cavity of the supply plate does not have a separation pore structure connected therein before being loaded with fluid. A composition including a desired concentration of drug can be loaded into the holding cavity of the supply plate. Then, the separation pore structure can be connected in the holding cavity so that the fluid in the holding cavity is divided into 96 separation pores, which are aligned with the openings in the transfer adapter and the holes of the 96-well multiwell plate. The connection of the separation pore structure in the holding cavity can result in a uniform distribution of the fluid volume in each separation pore.

[0201] For example, the separation hole structure can be connected in the holding cavity before the drug composition is loaded. The separation hole structure can include one or more separation wall grooves or gaps that fluidly connect two or more separation holes, and the drug composition can be loaded into one or more separation holes, allowing the composition to be evenly distributed between the fluidly connected separation holes.

[0202] For example, the separation pore structure may be configured such that each separation pore is fluidly connected to each other, and a composition including a drug may be delivered into one or more separation pores such that the composition is evenly distributed between each separation pore.

[0203] For example, the separation pore structure can be configured such that a first subset of separation pores (e.g., the first half) are fluidically connected to each other, and a second subset of separation pores (e.g., the second half) are fluidically connected to each other, but the first subset of separation pores is not fluidically connected to the second subset of separation pores. In such an example, a first composition comprising a drug at one concentration can be delivered to one or more pores of the first subset of separation pores such that the first composition is evenly distributed between each of the first subset of separation pores, and a second composition comprising a different concentration of drug (or different drugs) can be delivered to one or more pores of the second subset of separation pores such that the second composition is evenly distributed between each of the second subset of separation pores.

[0204] The fluid in the supply plate can be transferred to the 96-well multiwell plate containing the cells to be processed by centrifuging the transfer assembly under conditions that allow the fluid to pass from the separation wells of the supply plate through the openings in the transfer adapter and into the wells of the 96-well multiwell plate without substantially disturbing the cells attached thereto.

[0205] 3. Analysis: The effect of the drug on the cells in each well can be analyzed by live cell analysis. Bright field images or videos of the cells in each well can be obtained. If the cells are intrinsically fluorescent (e.g., due to a GFP gene transfected into the cell genome), fluorescent images of the cells can be obtained. Additionally or alternatively, the culture medium in each separate well can be retained for further testing using methods similar to those described in Example 1 above.

[0206] To observe specific activities of the cells in response to the drug, the medium in each well can be removed and the cells can be subjected to various assays using methods similar to those described above in Example 1. For example, in a staining assay, all cells in a 96-well multiwell plate can be processed and stained at once without the need for a robotic liquid handler or multichannel pipette. In a fluorescence assay, the fluorescent signal from the cells can be acquired.

Claims

1. A device for transferring a fluid, comprising: a planar sheet having a first planar surface, a second planar surface, and a plurality of openings, wherein each opening of the plurality of openings extends between the first planar surface and the second planar surface; a plurality of main extensions, wherein each main extension of the plurality of main extensions protrudes from the first planar surface and includes a main interior cavity, wherein each main extension is configured to reversibly couple to a feed hole of a feed plate to form a fluid-tight or leak-proof seal and to form a pocket between the main extension and an inner surface of the feed hole, and wherein the main extension includes structure configured to fluidly connect the pocket to the main interior cavity; as well as a plurality of secondary extensions, wherein each of the plurality of secondary extensions protrudes from the second planar surface and includes a secondary lumen, and wherein each secondary lumen is aligned with the opening and the primary lumen to create a continuous transfer lumen, and wherein each secondary extension is configured to reversibly couple to a receiving hole of a receiving plate, Wherein fluid entering the recess from the supply hole flows through the structure into the continuous transfer lumen and into the receiving hole.

2. The device according to claim 1, characterized in that Each of the plurality of main extensions has an inner cross-sectional area and an outer cross-sectional area, and the inner cross-sectional area is greater than the outer cross-sectional area.

3. The device according to claim 1 or claim 2, characterized in that: Each main extension has at least two regions, and wherein each of the at least two regions has a different angle relative to the planar sheet.

4. The device according to claim 3, characterized in that The at least two regions include a first region close to the flat sheet and a second region far from the flat sheet, and an angle of the first region relative to the flat sheet is greater than an angle of the second region relative to the flat sheet.

5. The device according to claim 1, characterized in that The structure includes an aperture, a hole, a slit, a gap, a notch, a groove or a channel.

6. The device according to claim 5, characterized in that The main extension includes four slots configured to be fluidly connected to the main lumen.

7. The device according to claim 6, characterized in that The slits are gaps in the periphery of the main extension.

8. A system for transferring a fluid, comprising: The device according to any one of claims 1 to 7; as well as a receiving plate, wherein the receiving plate is configured to be removably coupled to the second planar surface of the device and wherein each secondary extension is configured to be reversibly coupled to a receiving hole of the receiving plate, Wherein fluid entering the recess from the supply hole flows through the structure into the continuous transfer lumen and into the receiving hole.

9. The system according to claim 8, characterized in that The system further includes a supply plate including a hole or a plurality of holes, wherein the supply plate is configured to be removably coupled to the first planar surface of the device.

10. The system according to claim 9, characterized in that When the device and the supply plate are removably coupled, each opening of the plurality of openings is aligned with a different one of the plurality of holes.

11. The system according to claim 9, characterized in that The device comprises a plurality of main extensions, and wherein each of the plurality of main extensions is configured to seal against an inner surface of a different one of the plurality of holes of the feed plate.

12. The system according to claim 8, characterized in that The receiving plate includes a plurality of receiving holes, and wherein each of the plurality of openings aligns with a different one of the plurality of receiving holes when the device and the receiving plate are removably coupled.

13. The system according to claim 12, characterized in that The device includes a plurality of secondary extensions, and wherein each of the plurality of secondary extensions is configured to be inserted into a different one of the plurality of receiving holes.

14. The system according to claim 8, characterized in that The device comprises an adhesive on the first planar surface and / or the second planar surface.

15. The system according to claim 8, characterized in that The device includes a plurality of leaflets adjacent each of the plurality of openings, wherein at least one leaflet is movable between an open position and a closed position.

16. The system according to any one of claims 8 to 15, characterized in that: The device is configured to allow fluid to flow through the plurality of openings only when an external force is applied to the fluid.

17. A method for transferring a fluid, comprising: The device of any one of claims 1 to 7, a supply plate and a receiving plate to form a transfer assembly in a first position, wherein in the first position, the device is positioned below the receiving plate and above the supply plate; as well as centrifuging the transfer assembly about a rotation axis, wherein when the transfer assembly is centrifuged, the supply plate is positioned closer to the rotation axis than the device and the receiving plate, wherein the device comprises a plurality of openings and the feed plate comprises a plurality of holes, and wherein each of the plurality of openings is aligned with a different hole of the plurality of holes when the transfer assembly is in the first position and when the transfer assembly is centrifuged.

18. The method according to claim 17, characterized in that The method further includes separating the supply plate from the device.

19. The method according to claim 17 or claim 18, characterized in that When the supply plate and the device are coupled, a seal is formed between each of the plurality of holes and the device.

20. The method according to claim 19, characterized in that The seal allows fluid to flow from the plurality of apertures through the plurality of openings but prevents the fluid from flowing between apertures of the plurality of apertures.

21. The method according to claim 17, characterized in that Each well of the plurality of wells includes a target agent attached to the well, and a fluid.

22. The method according to claim 21, characterized in that The target agent remains attached to the wells after centrifugation.

23. The method according to claim 21 or claim 22, characterized in that The fluid is transferred to the receiving plate after centrifugation.

24. The method according to claim 17, characterized in that The receiving plate includes a plurality of receiving holes, and wherein each of the plurality of openings aligns with a different one of the plurality of receiving holes when the transfer assembly is in the first position and when the transfer assembly is centrifuged.

25. The method according to claim 24, characterized in that When the receiving plate and the device are coupled, a seal is formed between each of the plurality of receiving holes and the device.

26. The method according to claim 25, characterized in that The seal allows fluid to flow through the plurality of openings to the plurality of receiving holes, but prevents fluid from flowing between the receiving holes of the plurality of receiving holes.

27. A method of transferring a fluid from a well of a supply plate comprising a plurality of wells and / or transferring a fluid to a well of a supply plate comprising a plurality of wells using the apparatus of any one of claims 1 to 7, wherein at least one well comprises a plurality of target agents and fluids attached to the well, comprising: A fluid transfer force is applied to the supply plate, wherein the fluid transfer force has a simultaneous and substantially uniform effect on the plurality of target agents in the at least one well.

28. The method according to claim 27, characterized in that Each of at least two of the plurality of wells includes a plurality of target agents attached to the well and a fluid, and wherein the fluid transfer force has a simultaneous and substantially uniform effect on the plurality of target agents in each of the at least two wells.

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