Engineered tips for liquid handling devices and related materials for liquid and solid sample processing

Abstract

The present application relates to a tip, in particular a pipette tip for liquid handling, comprising at least one aperture at the distal end, which is arranged such that objects substantially in line with the longitudinal axis are not directly impacted by the liquid flow when using the tip for handling liquids. The present application also relates to an assembly comprising a tip according to the present application, which is connected to a liquid handling device, such as a micropipette or a syringe, either directly or via an adapter; a method for handling a sample with a liquid such that objects substantially in line with the longitudinal axis are not directly impacted by the liquid flow when using a tip according to the present application for handling liquids; a kit; and various uses in liquid handling and in handling a sample with a liquid.

Description

Invention Field

[0001] This application relates to a pipette tip, particularly a pipette tip for handling liquids, having at least one orifice at its distal end, the orifice being configured such that, when liquids are handled using the pipette tip, an object substantially collinear with its longitudinal axis is not directly impacted by the liquid flow. This application also relates to an assembly comprising the pipette tip of this application connected directly or via an adapter to a liquid handling device, such as a micropipette or syringe; a method for handling samples with liquids such that, when liquids are handled using the pipette tip of this application, an object substantially collinear with its longitudinal axis is not directly affected by the liquid flow; a reagent kit; and various uses in liquid handling and in handling samples with liquids. Background Technology

[0002] Organoids are a unique and emerging biological model in basic and applied research, showing great promise in drug screening and personalized medicine. In typical tissue culture laboratory settings, these organoids are prepared and tested in liquid or soft hydrogel suspensions, requiring regular changes of the culture medium or replenishment of growth factors. To avoid sample loss during such experiments (including but not limited to downstream staining and sample collection for omics studies), the flow of culture medium during the aspiration and dispensing of fluid using existing pipettes can affect the sample, often leading to unwanted aspiration or damage to micron-sized organoids. Similar challenges exist with other 3D samples such as animal embryos.

[0003] This less-than-ideal pipetting technique is a major obstacle to automation, especially for soft and solid samples in liquid solutions. Therefore, it is common practice in laboratories for researchers to manually aspirate and dispense culture media with extreme care to avoid aspiration, and to image and visually inspect the sample's presence and position within the container during the procedure. At the end of the procedure (often lasting several days), sample viability is significantly affected by the operator's experience and attentiveness.

[0004] Therefore, this artificial intervention limits the potential application of organoid systems in high-throughput experiments that provide statistical significance for relevant research. While liquid handling devices exist on the market, they employ traditional pipette tips and are designed to handle liquid biological samples, thus failing to account for the final solid 3D sample within the culture medium container, leading to the aforementioned problems. The lack of easily integrated devices and methodologies hinders the development of automated 3D sample handling schemes, and indeed the entire industry reliant on high-throughput research.

[0005] Organoids encompass a vast area of ​​interest in the life sciences, such as gene screening, disease modeling, the development of drugs for cancer and infectious disease pathology, clinical trials, and personalized medicine. Notably, in December 2022, the U.S. government approved the FDA Modernization Act 2.0. This act eliminates the need for animal testing to assess drug safety and efficacy. The EU's "alternative, reduction, and optimization" policy also encourages the gradual elimination of animal samples. Therefore, in vitro systems, such as stem cell-derived organoids, are at the forefront as biological models for drug testing.

[0006] However, both academia and industry still rely on tools designed for liquid biological samples to process these novel samples. Therefore, the proposed design for novel pipette tips perfectly aligns with market trends and the needs of both industry and academia. Our invention will not only improve the efficiency of manual organoid experimental protocols but also integrate into automated experimental protocols. In fact, these designs can convert many existing liquid handling robots into 3D sample processing devices with minimal effort. In turn, the automation of these organoid experimental protocols will enhance high-throughput research in both academia and industry.

[0007] US9733169B2 discloses a pipette tip device for dispersive SPE. The device includes a pipette tip with a lower baffle, a loose adsorbent that is freely movable during extraction, and a baffle system shaped to disrupt the flow of liquid sample aspirated into the pipette tip. The baffle system includes an insert that can be separable from or integrally formed with the interior of the pipette tip.

[0008] US 2014-0377147A1 discloses a chromatographic pipette tip comprising a first container and a second container, the second container having openings at two opposing ends and, in each example, including a bottom orifice through which sample liquid can be aspirated or expelled. The two containers are fluidically sealed to each other. A reaction matrix is ​​disposed in one of the containers, and when used as intended, the sample liquid flows through the reaction matrix by the push of a pipette connected to the chromatographic pipette tip. The sample liquid is aspirated and expelled by the chromatographic pipette tip from opposite directions and flows through the reaction matrix in only one direction.

[0009] US 2022-0152604 discloses a pipette tip for handling liquids, comprising a closed distal end of the tip and at least one lateral opening located above the closed end. In other embodiments, three or more lateral openings may be included, such as four to six lateral openings, provided that the number and / or size of these openings does not compromise the structural integrity of the tip. According to the specification, at least one embodiment of the attachment device can limit the rise in pressure within the test container during liquid dispensing. This is claimed to enable accurate and safe dispensing of liquid from the attachment device into the test container. Furthermore, when the device is immersed in the closed test container, the opening of the device can become blocked by the barrier material used to close the test container, or by crystalline or particulate reagents present in the test container. The closed opening of the attachment device will subsequently hinder the testing process because it may be difficult to release the liquid sample from the device. No intention to avoid sample aspiration or sample damage is disclosed or suggested.

[0010] WO 2015 / 156331A1 provides an attachment for liquid injection that allows for easy liquid injection into the inlet hole by simply connecting the pipette tip or pipette tip when injecting liquid into the inlet hole, while achieving excellent sealing performance and suppressing leakage during liquid injection without the risk of deformation or damage to the pipette or pipette tip.

[0011] WO 2020 / 127902A1 relates to a pipette tip extension attachable to a pipette tip. The pipette tip extension includes a proximal end, a distal end, and an outer wall extending between the proximal and distal ends. The outer wall has an outer side and an inner side, and a receiving aperture is formed at the proximal end for insertion of a pipette tip. The pipette tip extension further includes a bottom surface at the distal end, an inner cavity enclosed by the inner side of the outer wall and the bottom surface, one or more spacer elements disposed on the inner side of the outer wall and extending into the inner cavity, and a covering layer for interaction with fluid present in a fluid aspiration region.

[0012] WO 2020 / 132394A1 also relates to a pipette tip extension that can be attached to a pipette tip, comprising a proximal end, a distal end, and an outer wall extending between the proximal and distal ends. The outer wall has an outer side and an inner side, and forms a receiving aperture for insertion of the pipette tip at the proximal end and a dispensing aperture at the distal end. The pipette tip extension also includes an inner cavity, a spacer element connected to the inner side of the outer wall, and a constriction element. The constriction element (12) defines a stop (23) at the end of the pipette tip (20) and is configured as a fluid-permeable sieve structure.

[0013] Given the significant impact of organoid research, companies have attempted to provide solutions to the challenges mentioned above. However, most devices are only customized for the culture steps and offer little benefit in terms of general methods for handling 3D samples such as organoids. Existing solutions aim to modify the geometry of the culture plates or achieve gentle automated culture medium replacement, both of which require changes to the experimental procedures.

[0014] 3D samples, such as organoids, require frequent culture medium changes. Using existing tools may result in sample aspiration or damage, limiting throughput, reproducibility, and the implementation of automated solutions. Summary of the Invention

[0015] The purpose of this application is to provide an alternative for the liquid handling of biological samples, such as the gentle replacement of culture media for organoid samples, which is equally feasible in automated liquid handling systems. Other objectives and advantages will become apparent to those skilled in the art upon reading the more detailed description and embodiments below.

[0016] The above-mentioned objectives of this application are achieved by the claims appended.

[0017] In its first aspect, the aforementioned objective of this application is achieved by a pipette tip (1) for handling liquids, particularly a pipette tip (1), comprising a proximal end (p) and a distal end (d) and a longitudinal axis (I) extending therebetween, wherein a hole (2) is provided at the proximal end substantially concentric with the longitudinal axis (I) for connecting the pipette tip to a liquid handling device (e.g., a syringe or pipette), wherein the distal end (d) of the pipette tip is closed; and at least one hole (3) near the distal end. Its features are, At least one hole (3) located at the distal end (d) is configured such that when liquid is processed using the suction tip, an object substantially in line with the longitudinal axis is not directly affected and / or damaged by the liquid flow.

[0018] Preferably, the suction head (1) according to this application has at least one hole (3) at the distal end forming an angle of 45 to 180 degrees, preferably 90 to 180 degrees, with the longitudinal axis (I), and is preferably arranged in a substantially right-angled manner.

[0019] Conventional suction heads (such as) Figure 1 As shown in Figure A, it consists of a hollow cone with proximal and distal holes (in...). Figure 1In section A, these two holes are labeled “p” and “d” respectively. They are concentric and aligned with the longitudinal axis (I). With this design, the sample is directly affected and / or damaged by the flow of the liquid being processed (such as a medium). Therefore, the present inventors have proposed a different design in the context of this application, described in the context of several (four) general and different implementations (designs) that can be manufactured and used in different ways. Of course, other variations may exist without departing from the concept of this application when considering the overall disclosure of this application given herein. In general, the device proposed in this application decouples the orientation of the two holes. The proximal hole (2) remains substantially concentric with the longitudinal axis (I), maintaining compatibility with existing liquid handling equipment (such as micropipettes), while the distal hole (3) forms an angle of about 45 to 180 degrees with the axis, preferably about 90 to 180 degrees. In the process of dispensing liquids (such as media), in some designs, the liquid is injected into the container or vessel along the wall of the container or vessel to reduce turbulence. This reduces potential damage to samples (such as cells, cell clusters, or organoids) and their displacement, thereby shortening the time spent waiting for them to settle between consecutive operations. Furthermore, the fluid does not directly interact with the sample during aspiration. This reduced interaction significantly decreases the likelihood of sample aspiration (and loss).

[0020] The pipette tip design of this application allows for media replacement without moving the biological sample, thereby minimizing the interaction between the sample and the aspiration and dispensing flow. This design is fully compatible with existing laboratory instruments, equipment, and hardware. It is a low-cost device that can be easily integrated into existing laboratory procedures, automating the pipetting of soft and solid samples in liquid solutions with minimal exposure of the contained sample to the media flow. Therefore, this design minimizes sample loss or damage and improves the efficiency, speed, and success rate of biological experimental protocols. Finally, this design also supports and enables gentle media replacement, especially in automated processes.

[0021] Preferred is the pipette tip (1) of this application, wherein the distal end (d) of the pipette tip forms a closed distal end (3), or the distal end is closed by a separate plug (4). Preferred is the pipette tip (1) according to this application, wherein the distal end (d) of the pipette tip forms a crossbar or protrusion (5), which is configured substantially perpendicular to the longitudinal axis (I), preferably substantially circular, and / or preferably located at or off-center from the longitudinal axis (I). The crossbar or protrusion (5) may contain a channel (6) for the flow of the liquid being processed. The size and shape of the crossbar or protrusion (5) may also be designed to prevent the pipette tip from falling below a certain height after insertion into the sample container or dish. Thus, a sample capture area is formed by the crossbar (5) and the wall of the container or dish. Furthermore, this allows for limiting the amount of liquid that can remain in the container during the aspiration step. These capture areas also allow for understanding the position of the sample within the container without the need for additional tools such as computer vision or a microscope.

[0022] In its second aspect, the aforementioned objective of this application is achieved by an assembly comprising at least two or more pipette tips (1) of this application, integrated into or integrally formed on a support frame, bracket, plate, or cap, preferably arranged in a straight line and / or parallel along an axis, or concentrically arranged (e.g., in a circular embodiment), wherein the assembly is preferably configured as a support frame, bracket, plate, or cap, into which two or more pipette tips (1) are integrated. This aspect also relates to an assembly comprising a pipette tip (1) according to this application, which is attached directly or via an adapter (e.g., a commonly used pipette tip) to a device for handling liquids, such as a syringe or micropipette (e.g., a handheld pipette), or a pipette in a liquid handling workstation, such as a positive displacement pipette, particularly an air displacement pipette.

[0023] In its third aspect, the above-mentioned objective of this application is achieved by a method for processing samples with liquid, the method comprising the steps of: i) providing the pipette tip (1) or assembly of this application; ii) attaching the pipette tip (1) or assembly directly or via an adapter (e.g., a commonly used pipette tip) to a syringe or micropipette; iii) a) drawing liquid into the pipette tip (1) or assembly, and iv) a) discharging liquid from the pipette tip (1) or assembly into a suitable culture container containing the sample, or iii) b) drawing liquid from the culture container containing the sample into the pipette tip (1) or assembly, and iv) b) discharging liquid from the pipette tip (1) or assembly into a suitable dish or container, in such a manner that an object substantially aligned with the longitudinal axis is not directly affected and / or damaged by the liquid flow when processing liquid with the pipette tip (1) or assembly.

[0024] In its fourth aspect, the above-mentioned objective of this application is achieved by the use of the suction tip (1) or assembly of this application in handling liquids and / or processing samples with liquids, in such a way that an object substantially in line with the longitudinal axis is not directly affected and / or damaged by the liquid flow when using the suction tip (1) or assembly to handle liquids.

[0025] In its fifth aspect, the above-mentioned objective of this application is achieved by a kit for liquid-processed samples, comprising materials for carrying out the methods of this application, such as buffers or cell culture media selected for culturing and / or maintaining and / or storing one or more samples (particularly for 3D organoid culture, tissue culture, preservation of organs or portions thereof), reagents for processing the samples (preferably for performing cell lysis reactions, staining reactions, binding reactions, or for removing embedding media), and / or liquid in a transport fluid for collecting particles for further analysis; and also comprising pipette tips (1) and / or components according to this application.

[0026] As described above, the aforementioned objective of this application is achieved by a specific pipette tip (1), such as a pipette tip (1) for handling liquids (e.g., cell culture media). This pipette tip (1) has a generally elongated body, preferably substantially conical, comprising a proximal end (p) connected to, for example, a positive displacement device (e.g., a syringe or pipette) or a vacuum generating device (e.g., an air displacement pump, such as a piston-equipped syringe or micropipette), and a distal end (d) on the side of the container to be emptied, stirred, or filled. A longitudinal axis (I) extends between the proximal end (p) and the distal end (d).

[0027] The proximal orifice (2) is configured to be substantially concentric with the longitudinal axis (I) for connecting the pipette tip to a vacuum generating device, such as a pipette. The distal end (d) of the pipette tip is closed. The closure of the distal end can be achieved by any suitable means, for example, by forming the distal end (d) of the pipette tip with the distal end (3) closed during the manufacturing process of the pipette tip. Alternatively, the distal end can be closed by a separate plug (4).

[0028] The pipette tip includes at least one additional orifice (3) near its distal end. Importantly, this at least one orifice (3) at the distal end (d) is characterized in that it is configured such that, when handling liquids with the pipette tip, an object substantially aligned with the longitudinal axis is not directly affected and / or damaged by the liquid flow. The arrangement of the at least one orifice (3) at the distal end (d) allows liquids to enter or exit the container or dish as designed during dispensing and / or aspiration / removal, reducing turbulence. This reduces potential damage to samples (such as cells, cell clusters, or organoids) and their displacement, thereby shortening the time spent waiting for them to settle between successive steps. The design of this application enables the automation of pipetting soft and solid samples in liquid solutions with minimal exposure of the contained sample to the medium flow.

[0029] Therefore, it is preferred that the suction tip (1) according to this application has at least one hole (3) at its distal end forming an angle with the longitudinal axis (I), the angle being selected in such a way that an object substantially aligned with the longitudinal axis is not directly affected and / or damaged by the liquid flow when the suction tip is used to process liquid. The angle may be at least between about 20 and about 180 degrees relative to the longitudinal axis (I), at least between about 45 and about 180 degrees, preferably between about 90 and about 180 degrees, and preferably arranged at a substantially right angle. More preferably, as described above, the at least one hole (3) at its distal end is configured such that the liquid flow injected or drawn into the container or vessel via the suction tip moves along the wall of the container or vessel.

[0030] In an embodiment of the suction head (1) of this application, 1, 2, 3, 4, 5 or 6 holes (3) are provided at the distal end (d), for example, opposite each other (2 or 4 holes), at one-third of the circumference (3 holes), or in the shape of a pentagon (5 holes) or a star (6 or more holes).

[0031] In an embodiment of the suction head (1) of this application, a crossbar or protrusion (5) is formed at the distal end (d) of the suction head, which is configured substantially perpendicular to the longitudinal axis (I), preferably substantially circular, and / or more preferably disposed at the center or off-center of the longitudinal axis (I). Preferably, the crossbar or protrusion is substantially circular, elliptical, or semi-circular, and may also be disposed at the center or off-center of the longitudinal axis (I), i.e., extending further to one side than to the other. The design of the crossbar will take into account how to avoid turbulence in a given vessel or container shape when applying the medium.

[0032] Adding a physical barrier between the distal well and the sample further reduces the interaction between the medium and the sample (e.g., organoids). This barrier acts as a physical support for the upward passage of the distal well. The barrier also functions as a sample trap. The size and shape of the crossbar or protrusion (5) can be further designed to prevent the pipette tip from falling below a specific height after insertion into the sample container or dish. Thus, a sample trapping area is formed by the crossbar (5) and the wall of the container or dish. Furthermore, this allows for limiting the amount of liquid that can remain in the container during the aspiration step. These trapping areas also allow for understanding the position of the sample within the container without the need for additional tools such as computer vision or a microscope. All of these avoid the need for visual inspection, thereby reducing the requirements for implementing this method in an automated system.

[0033] In another embodiment of the suction head (1) of this application, the crossbar or protrusion (5) includes at least one channel (6) for the flow of the liquid being processed. In this embodiment, the flow of the medium is preferably at an angle of about 90 to about 180 degrees relative to the longitudinal axis (I), i.e., "upward" in the direction of the proximal end (p).

[0034] In the particularly preferred embodiment of this application, “Design 1” (see…) Figure 1 In B), the pipette tip (1) is sealed at the bottom, and the distal end hole (3) faces the side of the pipette tip, so that the medium flow will not directly impact or aspirate the sample / specimen. The design can preferably be used as a ready-to-use preparation pipette tip with the new design, or an existing pipette tip can be converted to the current new design by using benchtop equipment that can be used in the laboratory (similar to a glass needle pulling device, such as a Sutter instrument), closing the bottom hole and drilling the distal end hole on the side.

[0035] A particularly preferred embodiment of the suction head (1) of this application, "Design 2" (see [reference]). Figure 1 In C), similar to the previous design, the distal orifice (3) is located on the side of the pipette tip / towards the side of the pipette tip. This implementation is derived from conventional pipette tips by using a "gate" or plug (4) to close the bottom orifice. The lower part of the plug is a circular structure, the diameter of which can be designed to be equal to the diameter of the container at a selected height, forming a crossbar. With this feature, the pipette tip cannot reach below a selected height from the bottom of the container, thereby isolating the volume affected by the medium flow from the sample and preventing them from being unnecessarily squeezed.

[0036] In a particularly preferred embodiment of the suction head (1) of this application, “Design 3” (see Figure 1 D), the distal hole (3) faces the side of the suction head. Here, the bottom circular structure (bar) is integrated into the design. Figure 2 B shows an example of a 3D printed prototype mounted as an accessory on a conventional suction head.

[0037] Another particularly preferred embodiment of the suction head (1) in this application, "Design 4" (see [reference]). Figure 1 In E), the U-shaped channel spans or is integrated into the circular structure of the bottom crossbar, forming a distal orifice facing upwards towards the top of the container. Therefore, aspiration and dispensing are performed relative to the sample position in an opposite ("upward") direction, while the crossbar / bottom structure still provides additional protection for the sample.

[0038] Although this application is described herein as a standalone pipette tip, the concept of this application can also be applied to other pipettes, such as Pasteur pipettes with a rubber bulb attached to the proximal (p) end.

[0039] In a particularly preferred embodiment of the suction tip (1) of this application, the suction tip includes at least one lateral baffle (8) on the crossbar or protrusion (5) to guide the liquid flow, preferably in the direction of the hole (2) and / or substantially in the direction of the longest distance between the hole and the edge of the crossbar or protrusion (5). This further reduces any turbulence.

[0040] Generally, the material of the pipette tip (1) of this application can be any suitable material for pipette tips used for liquid handling (e.g., suitable for cell culture). The pipette tip can be a disposable (“single-use”) pipette tip or a reusable pipette tip, such as a washable stainless steel tip, glass or plastic, such as autoclaved PVC or PE.

[0041] All designs, especially designs 3 and 4, can be made into ready-to-use disposable tips, attachments for regular tips, or washable stainless steel or glass tips. The choice of implementation will depend on the application requirements (e.g., the need for sterility).

[0042] The pipette tip (1) according to this application is generally compatible with and can be used in existing liquid handling equipment, such as syringes or pipettes, such as air-displacement pipettes, especially micropipettes. Therefore, the overall shape is basically conical. Preferably, the pipette tip (1) of this application is configured to be ejected from a pipette or syringe (e.g., a handheld pipette or syringe, or a liquid handling workstation) by means of an ejection mechanism.

[0043] In another important embodiment of the suction head (1) of this application, the suction head further includes a crossbar or protrusion located on the upper part of the suction head and configured to be substantially perpendicular to the longitudinal axis (I), preferably substantially circular, which preferably creates a height positioning element or a lateral positioning element. Preferably, the suction head (1) of this application includes a height positioning element (9) and a lateral positioning element (10), wherein the lateral positioning element (10) is configured to be offset below the height positioning element (9).

[0044] The height locator (9) can be designed in size and shape to prevent the pipette tip from being inserted below a specific height inside the sample container or dish. In this way, a sample capture zone is created by the baffle (9) against the wall of the container or dish. Furthermore, this allows for limiting the amount of liquid that can remain in the container during the aspiration step. The lateral locator (10) ensures that the pipette tip (1) is correctly positioned inside the container or dish. All of these eliminate the need for visual inspection, reduce the requirements for implementing this method in automated systems, and ensure the repeatability of operations in different procedures.

[0045] Furthermore, the probe (1) according to this application is preferred, wherein the probe (1) further includes at least one gas exchange channel disposed in the height positioner (9) and / or the lateral positioner (10). This avoids the generation of excessively high or low pressure within the container.

[0046] As described above, the probe (1) of this application is used to process liquids. The type of liquid is not limited, depending on the intended purpose; preferably, the liquid is selected from: buffer solutions or cell culture media for culturing and / or maintaining and / or storing one or more samples, particularly for 3D organoid culture; reagents for processing samples, preferably for performing cell lysis reactions, staining reactions, binding reactions, or for removing embedding media; and transport solutions for collecting particles for further analysis. More preferably, cell culture media are used for culturing and / or maintaining and / or storing one or more biological samples (particularly for cell culture or 3D organoid culture purposes).

[0047] With the key implementation of the pipette tip (1) of this application, the above-mentioned objective of this application is achieved by a pipette tip that is directly or via an adapter (e.g., a standard pipette tip (7)) connected to a micropipette or syringe. The implementation of the pipette tip (1) of this application makes the pipette tip more compatible with the prior art and can be used directly by rubbing the pipette tip onto a standard standard pipette tip (7). Other adapters can also be used, but they seem less convenient.

[0048] Another aspect of this application relates to a component comprising a pipette tip (1) according to this application, which is connected directly or via an adapter (e.g., a conventional pipette tip) to a syringe or micropipette, such as a handheld pipette or a pipette in a liquid handling workstation, particularly an air-displacement pipette. This is another example of the compatibility of the pipette tip (1) of this application.

[0049] Another aspect of this application relates to a method for processing a sample with liquid, the method comprising the steps of: i) providing a pipette tip (1) or assembly according to the application; ii) attaching the pipette tip (1) or assembly directly or via an adapter (e.g., a commonly used pipette tip) to a syringe or micropipette; iii) a) drawing liquid into the pipette tip (1) or assembly, and iv) a) discharging liquid from the pipette tip (1) or assembly into a suitable culture container containing the sample, or iii) b) drawing liquid from the culture container containing the sample into the pipette tip (1) or assembly, and iv) b) discharging liquid from the pipette tip (1) or assembly into a suitable dish or container, in such a manner that an object substantially aligned with the longitudinal axis is not directly affected and / or damaged by the liquid flow when processing the liquid with the pipette tip (1) or assembly.

[0050] Preferably, the method according to this application includes an aspiration step performed as the pipette tip moves toward the bottom of the vessel or container (“descending”) and / or an expulsion step performed as the pipette tip moves toward the top of the vessel or container (“ascending”). This further reduces any influence of liquid on the sample.

[0051] As described above, in the method of this application, the arrangement of at least one orifice (3) at the distal end (d) allows the liquid to be injected into or aspirated from the container or vessel as designed during liquid discharge, reducing turbulence. This reduces potential damage to samples (such as cells, cell clusters, or organoids) and their displacement, thus shortening the time spent waiting for them to settle between successive steps. The design of this application enables the automation of pipetting of soft samples in liquid solutions with minimal exposure of the contained sample to the medium flow.

[0052] Referring to the above description, the preferred method of this application is wherein the culture container is a cell culture container (e.g., a 3D cell culture container), and the object is selected from cells (e.g., embryonic cells), patient-derived tissues, organs or parts thereof, embryos or organoids.

[0053] The pipette tip (1) and / or components according to this application can be used with any test tube, centrifuge tube, round-bottom tube or vial, centrifuge tube, tray or box container, microplate or container where a sample can be collected, for example, at its bottom and where liquid needs to be added or removed.

[0054] Another aspect of this application relates to the use of the pipette tip (1) or assembly of this application in handling liquids and / or treating samples with liquids, in such a way that an object substantially in a straight line with the longitudinal axis is not directly affected and / or damaged by the liquid flow when using the pipette tip (1) or assembly to handle liquids.

[0055] Another aspect of this application relates to a kit for processing liquids and / or processing samples with liquids, comprising materials for carrying out the methods of this application, such as buffers or cell culture media selected for culturing and / or maintaining and / or storing one or more samples (particularly 3D organoid cultures, tissue cultures, preserved organs or portions thereof), reagents for processing samples (preferably for performing cell lysis reactions, staining reactions, binding reactions, or for removing embedding media), and / or liquids in a transport fluid for collecting particles for further analysis; and also comprising pipette tips (1) and / or components according to this application. Another aspect of this application relates to the use of the above-described kit in processing liquids (e.g., cell culture media), or in processing samples with liquids according to the methods of this application.

[0056] Another aspect of this application relates to the use of the pipette tip (1) or components of this application in processing liquids (e.g., cell culture media) or in processing samples with liquids according to the methods of this application.

[0057] In the context of this application, the term "about" means a deviation of ±10% from a given value, unless otherwise stated.

[0058] The working principle of pipette tips in the prior art is as follows: Figure 3 As shown.

[0059] exist Figure 3 Image A shows the Akura™ 96 / 384 Spheroid Microplate (InSphero AG). This product comprises a multi-well plate with a specific geometry. The walls of the wells are angled, and the medium is aspirated from them using an angled pipette tip, which is claimed to ensure that organoids are not aspirated. Advantageously, this is a simple solution that can be integrated into existing culture protocols. Unfortunately, this technology focuses on the culture step, and each well contains only one spheroid. It is expensive (50 to 100 euros per plate). No alternative operating strategy is provided in cases where the spheroid needs to be removed from the plate.

[0060] exist Figure 3 In section B, the product Gri3D® (SUN bioscience SA) is shown. This product comprises a multi-well plate in which hydrogel forms micropores within individual wells. Advantageously, this is a high-throughput device. Media replacement is performed via side wells, thus avoiding sample contact. This product is expensive (€500 per plate). No alternative handling strategy is provided in cases where it is necessary to remove spheroids from the plate. Furthermore, the size of the spheroids depends on the plate used.

[0061] exist Figure 3 C shows an automated media changing module (Agilent Technologies, Inc.). Advantageously, this provides an automated module with a washable vertical pipette tip for changing media on a 96-well U-bottom multi-well plate, automating lengthy operations. The pipette tip is simply positioned to the side of the well, and slow media changing avoids disturbing the spheroids. However, only one spheroid can be processed per well. This device is dedicated to culture. No alternative operating strategy is provided for situations where spheroids need to be removed from the plate.

[0062] The embodiments of this application eliminate the problem of directional flow of the pipette tip. This provides a universal solution for manipulating cells, tissues, and / or organoids, regardless of their stage, size, and number of wells, and can be easily adapted to both manual and automated operation schemes.

[0063] The embodiments of this application can be implemented using the same manufacturing process as conventional pipette tips or pipette tips, resulting in low-cost devices that are readily available to any laboratory and compatible with existing micropipette standards and virtually any laboratory setup.

[0064] All disclosed inventive concepts and designs can be applied to both manual and automated processes. Furthermore, they can be applied to living 3D samples (e.g., embryos, organoids) as well as fixed 3D samples (patient-derived tissue sections, organoids, embryos, etc.).

[0065] By borrowing technology from existing conventional suction heads, additional functions can be added: • Positive displacement, for example, used to improve the precision of viscous and volatile media. • Conductive suction tips, for example, for liquid level detection • Impedance measurement, for example, used to detect the passage of particles • Use low-retention materials, such as those designed to minimize the amount of liquid remaining inside the pipette tip.

[0066] The design of this application can also be applied to the following scenarios: • The sample container contains only a single sample (typically used for handling live samples). • Multiple samples are placed in the same sample container (typically used for fixed sample operations).

[0067] This application relates to the following:

[0068] Item 1. A pipette tip (1) for handling liquids, particularly a pipette tip (1). It comprises a proximal end (p) and a distal end (d) and a longitudinal axis (I) extending therebetween. The orifice (2) at the proximal end, which is configured to be substantially concentric with the longitudinal axis (I), is used to connect the pipette tip to a liquid handling device, such as a syringe or pipette. Wherein, the distal end (d) of the suction tip is closed; and At least one hole (3) near the distal end. Its features are, At least one hole (3) at the distal end (d) is configured in such a way that an object substantially in line with the longitudinal axis is not directly affected and / or damaged by the liquid flow when the liquid is processed using the suction tip.

[0069] Item 2. The suction head (1) according to Item 1, wherein at least one hole (3) at the distal end forms an angle of 20 to 180 degrees, preferably 90 to 180 degrees, with the longitudinal axis (I), and is preferably arranged in a substantially right-angled manner.

[0070] Item 3. The suction head (1) according to item 1 or 2, wherein the distal end (d) of the suction head forms a closed distal end (3), or the distal end is closed by a separate plug (4).

[0071] Item 4. A suction tip (1) according to any one of items 1 to 3, wherein the distal end (d) of the suction tip forms a crossbar or protrusion (5) substantially perpendicular to the longitudinal axis (I), preferably in a substantially circular shape, which preferably creates a capture area for the sample and / or is located at or off-center from the longitudinal axis (I).

[0072] Item 5. The suction head (1) according to Item 4, wherein the crossbar or protrusion (5) contains a channel (6) for the flow of the liquid being processed.

[0073] Item 6. The suction head (1) according to item 4 or 5, wherein the suction head includes at least one lateral baffle (8) on the crossbar or protrusion (5) for guiding liquid flow, preferably substantially in the direction of the hole (2) and / or substantially in the direction of the longest distance between the hole and the edge of the crossbar or protrusion (5).

[0074] Item 7. A pipette tip (1) according to any one of items 1 to 6, wherein the pipette tip is configured to be ejected from, for example, a handheld pipette or syringe, or a pipette or syringe of a liquid handling workstation via an ejection mechanism.

[0075] Item 8. A pipette tip (1) according to any one of items 1 to 7, wherein the pipette tip is configured to be compatible with existing micropipettes or syringes, and preferably is generally conical in shape.

[0076] Item 9. A pipette tip (1) according to any one of items 1 to 8, wherein the pipette tip is connected directly or via an adapter such as a common pipette tip (7) to a micropipette or syringe.

[0077] Item 10. A suction head (1) according to any one of items 1 to 9, wherein the suction head further includes a crossbar or protrusion at the upper part of the suction head, which is configured to be substantially perpendicular to the longitudinal axis (I), preferably substantially circular, and preferably provides a height positioning element or a lateral positioning element.

[0078] Item 11. The suction head (1) according to Item 10, wherein the suction head (1) includes a height locator (9) and a lateral locator (10), wherein the lateral locator (10) is offset below the height locator (9).

[0079] Item 12. The suction head (1) according to Item 10 or 11, wherein the probe (1) further comprises at least one gas exchange channel disposed in the height positioner (9) and / or the lateral positioner (10).

[0080] Item 13. A pipette tip (1) according to any one of items 1 to 12, wherein the liquid is selected from buffer or cell culture media for culturing and / or maintaining and / or storing one or more samples (particularly for 3D organoid culture, tissue culture, preservation of organs or parts thereof), reagents for processing samples (preferably for cell lysis, staining, binding, or removal of embedding media), and / or transport fluids for collecting particles for further analysis, and / or viscous or volatile liquids or media.

[0081] Item 14. An assembly comprising at least two or more suction heads (1) of any one of items 1 to 13, integrated in a support frame, bracket, plate or cover, preferably arranged in a straight line and / or in parallel along an axis, wherein the assembly is preferably configured as a support frame, bracket, plate or cover, and two or more suction heads (1) are integrated into the support frame, bracket, plate or cover.

[0082] Item 15. An assembly comprising a pipette tip (1) of any one of items 1 to 13, which is attached directly or via an adapter (e.g., a conventional pipette tip) to a device for handling liquids, such as a syringe or a micropipette, such as a handheld pipette, or to a pipette attached to a liquid handling workstation, such as a positive displacement pipette, particularly an air displacement pipette.

[0083] Item 16. The nozzle (1) according to any one of items 1 to 13, or the component according to item 14 or 15, which is a disposable nozzle or component, or a reusable nozzle or component, such as a washable stainless steel nozzle or component.

[0084] Item 17. A method for processing a sample with liquid, the method comprising the steps of: i) providing a pipette tip (1) or assembly of any one of items 1 to 16, ii) attaching the pipette tip (1) or assembly directly or via an adapter (e.g., a conventional pipette tip) to a syringe or micropipette, iii) a) drawing liquid into the pipette tip (1) or assembly, and iv) a) discharging liquid from the pipette tip (1) or assembly into a suitable culture container containing the sample, or iii) b) drawing liquid from the culture container containing the sample into the pipette tip (1) or assembly, and iv) b) discharging liquid from the pipette tip (1) or assembly into a suitable dish or container, in such a manner that an object substantially in a straight line with the longitudinal axis is not directly affected and / or damaged by the liquid flow when processing the liquid with the pipette tip (1) or assembly.

[0085] Item 18. The method of Item 17, wherein the culture vessel is a cell culture vessel, such as a 3D cell culture vessel, and the object is selected from cells, patient-derived tissues, organs or parts thereof, embryonic cells, embryos and organoids.

[0086] Item 19. The use of any of the pipette tips (1) or components in the treatment of liquids and / or the treatment of samples with liquids in such a manner that an object substantially in line with the longitudinal axis is not directly affected and / or damaged by the liquid flow when the pipette tips (1) or components are used to treat liquids.

[0087] Item 20. A kit for processing liquids and / or processing samples with liquids, comprising a buffer or cell culture medium selected for culturing and / or maintaining and / or storing one or more samples (particularly for 3D organoid culture), and further comprising a pipette tip (1) or component of any one of items 1 to 16. Attached Figure Description

[0088] This application will be further described in the following embodiments with reference to the accompanying drawings; however, this application is not limited thereto. For the purposes of this application, all referenced documents are incorporated herein by reference in their entirety.

[0089] Figure 1 An overview of the conventional (A) and proposed (BE) pipette tip designs is shown, both in cross-sectional views. The pipette tip walls are dark gray, while the channels through which the medium flows are light gray. “p” and “d” indicate the proximal and distal orifices, respectively, while arrows indicate the direction of aspiration and dissipation. Yellow spheres represent samples in experiments, while blue borders indicate containers. Notably, in the conventional tip (A), the sample is directly displaced / affected by the liquid flow, while in all proposed designs (BE), the sample is not impacted by the medium flow.

[0090] Figure 2 Design 1 is shown in (A) (see Figure 1 (A) The prototype of Design 3 was made by sealing the distal end hole at the bottom of a conventional pipette tip and drilling a new side hole. (B) The prototype of Design 3 was made by 3D printing as an accessory for a conventional pipette tip. This accessory changed the conventional liquid flow direction to the side and added a circular structure to protect the sample.

[0091] Figure 3 The working principle of commonly used (A) Akura™ 96-well spherical microplate, (B) Gri3D® and (C) automatic media replacement module is shown.

[0092] Figure 4 The gastrula used in the experiment on the first sample container (Experiment 1) is shown before (A) and after (B) 15 manual media changes using the modified pipette tip of this application.

[0093] Figure 5 The gastrula used in the current experiment (Experiment 2) is shown before (A) and after (B) 20 automated media changes using the modified pipette tip of this application.

[0094] Figure 6 Design 3.1 is shown. Figure 1An improved implementation of one of the designs shown (Design 3). The distal orifice can be of different shapes (rectangular or circular), depending on which is more convenient for the manufacturing process (see front view). Lateral baffles are added on both sides of the distal orifice to limit the longest direction in which the fluid tends to flow towards the bottom baffle, thus minimizing the interaction with the sample placed below the pipette tip (see section CC diagram). When aspirating the medium, the pipette tip body should not contact the wall of the sample container. If they do, the medium between the pipette tip body and the sample container wall will not be aspirated due to surface tension, leading to inaccuracy. For this reason: i. An angle α ≠ 0 is set between the pipette tip body and the vertical direction (see side view); ii. A distance D ≠ 0 is set between the pipette tip body and the boundary of the bottom baffle (see detail view B).

[0095] Figure 7 A and B show the use of Figure 2 The results of Experiment 1 and Experiment 2 of Design B (Design 3) are shown in Figure 3.

[0096] Figure 8 Showing the use of Figure 6 The results of Experiment 3 of the design shown (Design 3.1).

[0097] Figure 9 The results of staining zebrafish embryos using a standard pipette tip (top row) vs. Design 3.1 (bottom row) are shown (Experiment 4). The staining exhibits the same characteristics in both cases, indicating that using a modified pipette tip does not affect the staining quality. Three different stains (DAPI, TBXTA, and SOX17) were used.

[0098] Figure 10 Show Figure 1 Other implementations of the designs shown (designs 3 and 4) may be considered because in some applications, a smaller and non-circular bottom baffle may be required.

[0099] Figure 11 Show Figure 1 Other embodiments of design 3 shown.

[0100] Figure 12 The suction tip of this application is shown as an accessory to a syringe. This application reduces the amount of air between the piston and the distal orifice. This, in turn, improves the accuracy of aspiration and dispensing operations, especially when using viscous or volatile media.

[0101] Figure 13 A suction head suitable for a perforated plate is shown as... Figure 1An alternative design for the pipette tip. This design is suitable for specific sample containers, currently the widely used 96-well plate in organoid experimental protocols, but can also be adapted to other containers (e.g., centrifuge tubes). The plate features protrusions that insert into the wells to ensure a predetermined lateral position (x and y coordinates). The protrusions also ensure a predetermined vertical position (height, z coordinate), preventing the pipette tip from being inserted into the wells, thus avoiding sample compression and maintaining a predetermined dead volume. Channels ensure gas exchange during media aspiration / dispensing. These features allow the pipette tip to maintain the same positioning conditions for all samples, thereby ensuring and improving the reproducibility of media replacement procedures.

[0102] Figure 14 Another embodiment of a multi-well plate adapter is shown. Multichannel pipette accessories can be manufactured to have the same features as the previous ones. Figure 13 The same features ensure repeatability and correct positioning of the pipette tips, improve experimental throughput, and decouple operation from operator experience in other human factors. In this implementation, the eight units of Design 3.1 are combined together to add positioning features (in the lateral and longitudinal directions). These eight units are designed to fit a standard 96-well multiwell plate, a design that can be adapted to any container potentially with multiple sample compartments.

[0103] Figure 15 A multiwell plate cap for standardized media changing is shown. By further multiplication, for example targeting 96 repetitions, the invention of the current embodiment provides a newly designed cap for multiwell plates as an alternative to multichannel pipette tip accessories. With this cap, all tips are positioned in the same manner, thereby enhancing standardization and reducing human error in media exchange operations. The cap can be used only at the point where media change is needed, or, if made of autoclavable material, can remain inserted throughout the entire organoid culture experimental protocol. In this case, an additional standard cap can be placed on top to prevent media evaporation. The cap provides gas exchange channels and liquid change channels. In the latter, the operator can insert a conventional aspiration / dispensing device. This could be a conventional pipette (for serial media changing) or a set of tubing connected to a pump or valve, allowing media to be changed simultaneously in all wells in parallel. The purpose of this cap is to ensure that media changing is performed in the same manner and under the same conditions in all different wells. Similarly, the cap can be adapted to other sample containers and can accommodate different tip designs. Detailed Implementation Example The modified pipette tip used in this application

[0104] For the following concept-validation experiments, the same equipment was used. The sample containers were 1.5 ml centrifuge tubes. The modified pipette tips were 3D printed as an accessory to standard 1 ml pipette tips. The modified tips orthogonally separated the proximal and distal wells, creating a baffle at the bottom (see [link to documentation]). Figure 1 The same procedure was performed using a standard 1 ml pipette tip for comparison. Experiment 1: Manual replacement of culture medium

[0105] Experimental procedures using modified pipette tips: 1. Ninety-five 3D in vitro models (fixed embryonic organoids, such as gastrula) were divided into two groups (one group with 70 gastrula and the other with 25), imaged, and counted. Figure 4 A). 2. Transfer each group to a different sample container containing 0.5 ml of culture medium (5×SSCT). 3. For each container, use the modified pipette tip: a. Insert the pipette tip vertically, centering it in the container. As the liquid volume decreases, perform aspiration by lowering the pipette tip. b. Aspirate the culture medium, leaving the sample in a small amount of the medium. c. Discard the aspirated culture medium. d. Add 0.5 ml of fresh culture medium (5×SSCT). 4. Repeat steps 3a-c 15 times to complete 15 culture medium replacements. 5. Re-image and count the contents of each sample container. Figure 4 B).

[0106] Experimental procedures using standard pipette tips: 1. The 93 3D in vitro models (fixed gastrulations) were divided into two groups (one group of 71 gastrulations and the other group of 22), and were imaged and counted. 2. Use only a standard 1 ml pipette tip and perform the same procedure as described above. 3. After 15 culture medium changes, the contents of each sample container were imaged and counted again.

[0107] result: 1. Modified pipette tip: After 15 culture medium changes, the final total number of gastrula was 94, which is 98.95% of the original gastrula (one gastrula was transferred from the first tube to the second tube). 2. Standard pipette tips: After 15 culture medium changes, the final total number of gastruloids was 49, which is 52.69% of the original gastruloids. 3. See Table 1 (below) and Figure 7 A's summary.

[0108] Comparison of standard and modified suction heads: 1. Standard pipette tips: a. The results always depend on the researchers' experience and their attention in each step of the operation. b. In our laboratory, these procedures are routinely performed, and for researchers with average experience, sample loss after multiple culture medium changes is between 10-40%. c. Researchers need to introduce an angled tilt between the pipette tip and the container to reduce the resulting turbulence, visually inspect the movement of the samples, and ultimately use a microscope to actively prevent them from being sucked in. d. After dispensing new culture medium, additional time is required for the sample to settle to the bottom of the container due to the turbulent movement of the sample with the liquid. 2. The pipette tip according to this application: a. During culture medium aspiration, the pipette tip is inserted vertically into the sample container without introducing an angle of tilt. b. During the aspiration process, no special attention was paid to the movement of the gastrula, nor was their aspiration actively avoided to simulate a quasi-blind operation. c. During culture medium dispensing, the liquid is injected into the container along the container wall as designed, reducing turbulence. This reduces potential damage to the samples and movement, thus shortening the time spent waiting for them to settle between consecutive steps. d. This operation was performed by an untrained person who had performed similar operations (multiple culture medium changes for floating samples) fewer than 10 times prior to the current experiment. Therefore, this new design reduces the dependence of the results on the operator's experience and attention. Experiment 2: Automated Culture Medium Replacement

[0109] Experimental procedures using modified pipette tips: 1. Imaging and counting were performed on 93 3D in vitro models (fixed gastrulae). Figure 5 A), and transfer it to a sample container containing 0.5 ml of culture medium (5×SSCT). 2. On this container, use modified pipette tips and automated equipment: a. Insert the pipette tip vertically and center it in the container. As the liquid volume decreases, perform aspiration by lowering the pipette tip. b. Aspirate the culture medium, leaving the sample in a small amount of the medium. c. Discard the aspirated culture medium. d. Add 0.5 ml of fresh culture medium (5×SSCT). 3. Repeat steps 2a-c 20 times to complete 20 culture medium replacements. 4. Re-image and count the contents of the sample container. Figure 5 B).

[0110] Experimental procedures using standard pipette tips: 1. Image and count 92 3D in vitro models (fixed gastrulations). 2. Using standard 1 ml pipette tips, perform the same procedures as described above on an automated machine. 3. After 20 culture medium changes, the contents of the sample container were imaged and counted again.

[0111] result: 1. Modified pipette tip: After 20 culture medium changes, the final total number of gastrulations was 93, that is, 100% original gastrulations. 2. Standard pipette tips: After 20 culture medium changes, the final total number of gastruloids was 58, which is 63.01% of the original gastruloids. 3. For a summary of the comparisons, see Table 1 (below) and Figure 7 B. Table 1: Summary of experimental results, comparing the use of modified and conventional pipette tips in manual and automated culture medium exchange processes.

[0112] Comparison of conventional pipette tips with the tips of this application: 1. Standard pipette tips: a. The machine is a simple liquid processor, meaning it cannot process 3D samples. b. The sample is aspirated. 2. The suction head according to this application: a. Enables automated 3D sample processing. b. Transfer all the advantages of automation to this field: i. The results do not depend on human factors (attention, experience, etc.). ii. High throughput can be achieved. iii. Researchers can spend more time on experimental design and results interpretation, and less time on lengthy tasks. c. Avoid over-modification (no need for complex imaging systems to guide pipette tips). d. The liquid processor can be transformed into a "3D sample processor". Experiment 3: Using according to Figure 6 (Design 3.1) of this application

[0113] The purpose of this experiment was to simulate routine culture procedures on a 96-well multi-well plate, with one sample per well, by performing culture medium replacement on a single sample. The design changes shown (e.g., angles and distances for improved aspiration accuracy) can also be adapted accordingly. Figure 1 Design 4(E) is shown.

[0114] Experimental Protocol: Sample containers (96-well multi-well plates with a U-shaped bottom) were provided, with one single organoid per well. 150 µL of culture medium was removed from each well, and another 150 µL was dispensed (refilled). The culture medium replacement was repeated 10 times across the entire plate. In an automated environment, the results were recorded and compared with those from Design 3.1. Figure 8 Using a standard pipette tip, the average loss per repetition is ~14% of organoids. Using the Design 3.1 pipette tip, the average loss per repetition is 1% of organoids. Experiment 4: Using according to Figure 6 (Design 3.1) of this application

[0115] The purpose of this experiment is to demonstrate that the modified pipette tip does not affect the staining results when manually staining zebrafish embryos. Figure 9 This illustrates a comparison between a standard pipette tip and Design 3.1. Staining results show the same characteristics in both cases, indicating that the use of a modified pipette tip does not affect staining quality. This application's embodiment of the perforated plate cover ( Figure 15 Application in standardized culture medium replacement

[0116] Possible experimental protocols for changing the culture medium for developing organoids that require a controlled incubator environment: The modified cap is always placed over the multiwell plate: Cells are seeded into the multiwell plate, and the modified cap is applied. The original multiwell plate cap is placed on top, and the multiwell plate is placed in an incubator. For culture medium replacement, the multiwell plate is moved to a sterile hood. The original multiwell plate cap is removed, and using a regular pipette tip, the tip is inserted into the fluid exchange channel of the modified cap. The culture medium is aspirated and discarded. Then, a fresh culture medium is aspirated from the culture medium source using a regular pipette tip. The pipette tip is inserted into the fluid exchange channel of the modified multiwell plate cap, and the fresh culture medium is dispensed into the well. The aspiration and dispensing steps can be repeated for all wells. Afterward, the original multiwell plate cap is placed on top of the modified cap, and the multiwell plate is placed in an incubator until the next culture medium replacement operation.

[0117] Possible experimental protocols for changing the culture medium for developing organoids that need to be kept in a controlled incubator environment are shown below: Apply the modified cap to the multiwell plate only during culture medium changes: Seed cells in the multiwell plate and apply the original multiwell plate cap on top. Place the multiwell plate in an incubator. For culture medium changes, remove the multiwell plate to a sterile hood. Remove the original multiwell plate cap and apply the modified cap. Using a regular pipette, insert the tip into the fluid exchange channel of the modified cap. Aspirate the culture medium and discard it. Then, replace the pipette tip with a regular pipette tip and obtain fresh culture medium from the culture medium source. Insert the pipette tip into the fluid exchange channel of the modified multiwell plate cap and dispense the fresh culture medium into the wells. The aspiration and dispensing steps can be repeated for all wells. Afterward, remove the modified cap and place the original multiwell plate cap back on the original multiwell plate, then place the multiwell plate in an incubator until the next culture medium change.

[0118] List of reference numerals (p) Proximal end (d) Remote (I) Longitudinal axis (1) Pipette tip (2) Proximal hole (3) Distal hole (4) Stopper (5) Horizontal bar or protrusion (6) Channel (7) Common / routine pipette tips (8) Side baffle

Claims

1. A pipette tip (1) for liquid handling, particularly a pipette tip (1), comprising: The proximal end (p) and the distal end (d), and the longitudinal axis (I) extending between them. A hole (2) located at the proximal end, substantially concentric with the longitudinal axis (I), is used to connect the pipette tip to a liquid handling device, such as a syringe or pipette, wherein the distal end (d) of the pipette tip is closed; and At least one hole (3) near the distal end. Its features are, The at least one hole (3) located at the distal end (d) is configured such that when the liquid is processed using the suction tip, the object substantially in line with the longitudinal axis is not directly affected and / or damaged by the liquid flow.

2. The suction head (1) according to claim 1, wherein the at least one hole (3) located at the distal end forms an angle of 20 to 180 degrees, preferably 90 to 180 degrees, with the longitudinal axis (I), and is preferably arranged in a substantially right-angled manner.

3. The suction head (1) according to claim 1 or 2, wherein the distal end (d) of the suction head is formed with a closed distal end (3), or the distal end is closed by a separate plug (4).

4. The suction tip (1) according to any one of claims 1 to 3, wherein the distal end (d) of the suction tip is formed with a crossbar or protrusion (5) configured substantially perpendicular to the longitudinal axis (I), preferably substantially circular, preferably creating a sample capture area and / or disposed at or off-center from the longitudinal axis (I), wherein preferably the crossbar or protrusion (5) includes a channel (6) for the flow of the liquid being processed, and / or wherein the suction tip preferably includes at least one lateral baffle (8) on the crossbar or protrusion (5) to guide the liquid flow, preferably substantially in the direction of the orifice (2) and / or substantially tending toward the direction of the furthest distance between the orifice and the edge of the crossbar or protrusion (5).

5. The pipette tip (1) according to any one of claims 1 to 4, wherein the pipette tip is configured to be ejected from a pipette or syringe by, for example, a handheld pipette or syringe or a liquid handling workstation ejection mechanism, and / or wherein the pipette tip is configured to maintain compatibility with existing micropipettes or syringes, and preferably is generally conical in shape.

6. The suction head (1) according to any one of claims 1 to 5, wherein the suction head further includes a crossbar or protrusion at the upper part of the suction head, which is configured to be substantially perpendicular to the longitudinal axis (I), preferably substantially circular, and preferably creates a height positioning element or a lateral positioning element, wherein the suction head (1) preferably includes a height positioning element (9) and a lateral positioning element (10), wherein the lateral positioning element (10) is staggered below the height positioning element (9).

7. The suction head (1) according to claim 6, wherein the suction head (1) further comprises at least one gas exchange channel disposed in the height positioning element (9) and / or the lateral positioning element (10).

8. The pipette tip (1) according to any one of claims 1 to 7, wherein the liquid is selected from buffers or cell culture media for culturing and / or maintaining and / or storing one or more samples, particularly for 3D organoid culture, tissue culture, preservation of organs or parts thereof; reagents for processing samples, preferably for performing cell lysis reactions, staining reactions, binding reactions, or for removing embedding media; and / or transport fluids for collecting particles for further analysis; and / or viscous or volatile liquids or media.

9. An assembly comprising at least two or more suction heads (1) of any one of claims 1 to 8 integrated into a support frame, bracket, plate or cover, preferably arranged along an axis and / or in parallel, wherein the assembly is preferably configured as a support frame, bracket, plate or cover, and two or more suction heads (1) are integrated into the support frame, bracket, plate or cover.

10. An assembly comprising a pipette tip (1) according to any one of claims 1 to 8, the pipette tip (1) being attached directly or by means of an adapter such as a conventional pipette tip to a device for liquid handling, such as a syringe or a micropipette, such as a handheld pipette, or to a pipette attached to a liquid handling workstation, such as a positive displacement pipette, particularly an air displacement pipette.

11. The suction head (1) of any one of claims 1 to 8 or the component of claim 9 or 10, wherein it is a disposable suction head or component, or a reusable suction head or component, such as a washable stainless steel suction head or component.

12. A method for treating a sample with a liquid, the method comprising the following steps: i) providing the pipette tip (1) or assembly according to any one of claims 1 to 11, ii) attaching the pipette tip (1) or assembly directly or via an adapter such as a conventional pipette tip to a syringe or micropipette, iii) a) drawing liquid into the pipette tip (1) or assembly, and iv) a) dispensing the liquid from the pipette tip (1) or assembly into a suitable culture container containing a sample, or iii) b) drawing liquid from a culture container containing a sample into the pipette tip (1) or assembly, and iv) b) dispensing the liquid from the pipette tip (1) or assembly into a suitable container or dish, in such a manner that an object substantially in a straight line with the longitudinal axis is not directly affected and / or damaged by the liquid flow when the liquid is processed using the pipette tip (1) or assembly.

13. The method of claim 12, wherein the culture container is a cell culture container, such as a 3D cell culture container, the object being selected from cells, patient-derived tissues, organs or parts thereof, embryonic cells, embryos and organoids.

14. Use of the pipette tip (1) or assembly according to any one of claims 1 to 11 in liquid handling and / or sample handling with liquid, in such a manner that an object substantially in a straight line with the longitudinal axis is not directly affected and / or damaged by the liquid flow when the liquid is handled using the pipette tip (1) or assembly.

15. A kit for processing liquids and / or processing samples with liquids, comprising a liquid selected from buffers or cell culture media for culturing and / or maintaining and / or storing one or more samples, particularly for 3D organoid culture, and further comprising a pipette tip (1) or component according to any one of claims 1 to 11.

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