New multi-functional fluidic device for clamping biopsies

Abstract

To provide improved devices for use as tissue sample carrier in biopsy procedures in general.SOLUTION: A fluidic device 1 comprises a flow chamber 2 for accommodating a biological specimen on a carrier portion 3, and at least one flow channel 4a, 4b, 4c, 4d fluidly connected to the flow chamber 2. The fluidic device 1 has a layered structure comprising a bottom plate 5, a cover plate 6 and an insert 7 therebetween. The insert 7 comprises the carrier portion 3, and a frame portion surrounding the carrier portion 3, and is elastomeric to enable a biological specimen to be clamped inside an incision in the carrier portion 3.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a device for holding a biological specimen in a fixed orientation and / or for supplying a fluid to a biological specimen. The present invention further relates to an insert for such a device, and to a kit comprising such an insert and at least one cover plate which together with the insert may provide a device of the present invention. The present invention further relates to a system comprising an insert or device of the present invention. The present invention further relates to the use of the insert, kit or device for holding a biological specimen in a fixed orientation and / or for supplying a fluid to a biological specimen. [Background technology]

[0002] A prostate biopsy is a procedure in which a small tissue sample (biopsy) is taken from a patient's prostate to test for the presence or absence of prostate cancer. In most cases, a hollow needle is used to isolate a cylindrical sample with a diameter of about 1 mm and a length of, for example, 5-20 mm. The sample is usually H&E stained for histological examination under a microscope to determine the presence or absence of cancer cells and to evaluate the microscopic features and / or Gleason score of any cancer found. From the results, clinical risk can be determined, which can then be used to indicate appropriate treatment.

[0003] To prepare tissue samples for histological examination, decay due to autolysis or putrefaction must be prevented. This is done by fixation, which stops ongoing biochemical reactions and can increase the mechanical strength or stability of the tissue sample. The tissue sample is then embedded in paraffin or frozen to preserve tissue morphology and provide sufficient support for the tissue sample during sectioning. The usual fixative for tissues embedded in paraffin is neutral buffered formalin. These processes require the tissue sample to be in contact with many different fluids, so it is desirable to simplify the delivery of fluids to the tissue sample.

[0004] Although some fluidic devices can be anticipated in the above desires, the use of such devices for the preparation of tissue samples is fraught with difficulties. For example, if a tissue sample is placed on the surface of the flow chamber of such devices, such surfaces generally impair the perfusion of fluids and reduce the transparency required for microscopic examination. On the other hand, if the biopsy is allowed to move freely in the flow chamber, its orientation will be lost after a certain time, which may hinder accurate investigation of the most diagnostically valuable areas or the 3D structure of the tissue sample. Furthermore, an essential part of the biopsy collection process requires that the biopsy is first placed on the tissue, then picked up by a sponge and then placed in a vial. In order to be able to recognize how the biopsy is oriented after collection, ink needs to be used. Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, there is a need for improved devices for use as tissue sample carriers throughout biopsy procedures. [Means for solving the problem]

[0006] The present invention aims to at least partially satisfy the above-mentioned needs.

[0007] According to a first aspect of the present invention, there is provided a fluidic device as claimed in claim 1, in order to address the aforementioned objectives.

[0008] The fluidic device comprises a flow chamber for accommodating a biological specimen on a carrier part and at least one flow channel fluidly connected to the flow chamber. The fluidic device has a layered structure including a bottom plate, a cover plate and an insert therebetween. This means that the insert is sandwiched between the bottom plate and the cover plate. Furthermore, the insert comprises a carrier part and a frame part surrounding this part. The carrier part is preferably, but not necessarily, characterized as being elastomeric. Thus, the carrier part comprises or consists of an elastomeric material. Optionally, the frame part or the entire insert may also comprise or consist of a further elastomeric material. Preferably, the elastomeric material and the further elastomeric material are the same.

[0009] The carrier portion may have the shape of a membrane having a thickness that is much smaller than its width and length, and thus the membrane may be stretchable, i.e. elastic.

[0010] The term "elastomeric" as used herein means to exhibit mechanical properties of an object that allow it to be stretched so that it returns to its original shape and / or size when the stretching force is released. The mechanical properties are in particular such that the object exhibits rubber-like elasticity according to the IUPAC definition of "elastomer" meaning an elastomeric material. Thus, the phrases "elastomeric insert", "insert that is elastomeric" and synonymous expressions characterize mechanical properties that allow the insert to be stretched (i.e., reversibly stretchable) so that it returns to its original shape and / or size when the stretching force is released. Furthermore, an insert should be understood to be elastomeric if the insert is reversibly stretchable in at least one region of the insert, thereby allowing it to clamp a biological specimen, as further described below. This region may be present in or constituted by the carrier portion, and / or may be constituted by a region adjacent to the carrier portion that extends in the opposite direction from the peripheral boundary of the carrier portion through the frame portion. However, it is desirable to be able to have both uniform perfusion so that the delivered fluid reaches the entire surface of the biopsy and the orientation of the biopsy is maintained. Fluidic channels can therefore be used. The biopsy generally has the area of ​​greatest diagnostic value. Ideally, the specimen is oriented so that all diagnostically valuable layers are on the microtome slice for the pathologist's evaluation. Moreover, it is even more desirable to be able to analyze the sample directly in the fluidic device to facilitate handling and the number of manual steps.

[0011] As mentioned above, the inserts, as well as the fluidic devices, kits and systems including such inserts, are intended to clamp the biological specimen through the presence of an incision for receiving the biological specimen. Thus, the carrier part may have such an incision or may have an area intended (or adapted for) making such an incision before use. For example, this area may have a relief or color or other indicator that guides the user in making the incision (e.g. with a cutting device). The advantage of having a pre-prepared incision on the carrier part is that it eliminates the need for the user to make such an incision, thus facilitating the use of the insert, while the advantage of not having a pre-formed incision is that the incision may be adjusted to fit the size of the biological specimen to be received.

[0012] The insert may be a biopsy containing insert for use in a biopsy containing fluidic device.

[0013] As used herein, the term "biological specimen" is intended to mean one or more tissues, organisms or parts thereof (e.g., biopsy specimens), where the biological specimen is obtained from any of a variety of organisms. Exemplary organisms include, but are not limited to, mammals such as rodents, mice, rats, rabbits, guinea pigs, ungulates, horses, sheep, pigs, goats, cows, cats, dogs, primates (i.e., humans or non-human primates), and the like. Preferably, the biological specimen is a biopsy. To clamp the biological specimen, the insert is elastomeric so that it can be reversibly stretched (pushed, pulled, and / or bent). Additionally, the insert is modifiable so that it can be advantageously used to pick up, clamp, and / or fix in place the biological specimen. This can be done by a variety of approaches, some of which are illustratively described below.

[0014] Common to these approaches, an incision is provided in the part, preferably as a full cut-through through the entire thickness of the layer of the microporous carrier part, and the biological specimen is placed between the incisions by applying an external force to the insert. Incision as used herein means a slit-shaped opening that leaves a gap between the opposing cut ends of less than 1000 μm, preferably less than 500 μm, more preferably less than 250 μm, most preferably less than 100 μm, when no external force is applied to the insert. The incision preferably has a straight or cross shape. By applying an external force to the insert, the gap is widened so that it can accept the biological specimen. The external force may be applied as a bending force, a pulling force or a pushing force. Due to the elastomeric properties, the incised insert can clamp and / or fix the biological specimen in place when no external force is applied to the insert.

[0015] According to one approach, the biological specimen is placed between the incisions while a pressing force is applied to the carrier part, directly above or close to the incision in the direction of the layer thickness (i.e. from the top or bottom, relative to the orientation of the insert in use). The opening can be widened, for example with the help of tweezers, which can at the same time serve to place the biological specimen between the incisions. When the pressing force stops, the insert takes its original shape and the opening closes. This approach has the advantage that the insertion of the biological specimen can be automated and parallelized using existing fluidic device holders as disclosed herein.

[0016] According to another approach, the biological specimen is placed between the incisions while the opposing edges of the insert are pulled away from each other such that the opening of the carrier part is forced to widen. By ceasing the pulling force, the biological specimen is clamped between the incisions. According to yet another approach, the insert is bent along an axis extending through the carrier part such that the opposing edges of the insert approach each other along a circular trajectory, and the cut ends spread apart, leaving a widened wedge-shaped opening. When the bending force is ceased, the insert assumes its original shape and the opening closes. Thereby, a forceps-like clamping mechanism is created, which can be used to clamp and / or fix the biological specimen in place in a corresponding manner as described above, as well as serve to pick up the biological specimen.

[0017] When the biological specimen is securely clamped, the correct orientation of the biological specimen can be maintained, chemicals and nutrients can have free access to the biological specimen from above and below, and free optical observation for microscopic examination can be provided.

[0018] Preferably, the carrier portion contains holes or pores to allow fluid to pass through the carrier portion.

[0019] The fluidic device disclosed herein is particularly useful for performing experiments on biopsies, such as prostate biopsies. As mentioned in the introduction, the biopsy has the most diagnostically valuable area, and ideally the specimen is oriented so that all diagnostically valuable layers are on the microtome slice for evaluation. Although orientation is not achievable when a biological specimen such as a prostate needle biopsy is contained in a pre-filled formalin container, orientation can be maintained by immobilization (i.e. clamping) with an insert forming part of the fluidic device as disclosed herein, while perfusion is allowed. This can also be improved by providing holes, preferably micropores, in the carrier part. In this way, perfusion can also occur through the pores or micropores. Thus, the biopsy can be treated with multiple different fluids, including, for example, nutrients, chemotherapeutic agents, drugs, stains, and agents for preparing the biopsy, such as formalin and paraffin, in the same fluidic device. This fluidic device can further be used as a holder for a microtome for making slices for digital pathology and / or for preparing 3D images of the biopsy. As a further advantage, the fluidic devices disclosed herein have a simple design, are easy to manufacture, and are inexpensive.

[0020] The fluidic devices disclosed herein are preferably, but not necessarily, microfluidic devices, which are understood to deal with the manipulation of small, geometrically confined fluids on the sub-centimeter or sub-millimeter scale, where capillary permeation may dominate mass transport.

[0021] The carrier portion includes or consists of an elastomeric material, and / or the frame portion includes or consists of an additional elastomeric material. An elastomer as understood herein is a polymer that exhibits rubber-like elasticity, as defined by IUPAC. The elastomer provides elasticity to reversibly stretch (i.e., pull, push or bend) the insert, as disclosed herein. The elastomer may also be based on two or more polymers, each of which may be independently selected from homopolymers, copolymers and polymer blends.

[0022] Preferred elastomers are based on at least one polymer selected from the group consisting of thermoplastic elastomers (TPEs) and elastomers such as silicone and polybutadiene rubber, where particularly preferred elastomers are described in more detail in PCT application WO 2019 / 015988, which is incorporated herein by reference in its entirety, in particular for the purpose of disclosing elastomers for forming inserts (called membranes in WO 2019 / 015988) of fluidic devices. As used herein, the phrase "based on at least one polymer" describes an elastomer in which the at least one polymer is included as a major component. In some embodiments, the at least one polymer accounts for at least 60% by weight, at least 70% by weight, at least 80% by weight, at least 85% by weight, at least 90% by weight, at least 95% by weight, or at least 98% by weight, based on the total weight of all components included in the microporous carrier portion. In some embodiments, the frame portion and / or the microporous caylar portion consist substantially or consist entirely of the at least one polymer.

[0023] Preferred thermoplastic elastomers are selected from the group consisting of styrene block copolymers (TPS), preferably styrene ethylene butylene styrene block copolymers (SEBS), and thermoplastic polyurethanes (TPU). Particularly preferred elastomers are characterized by elasticity and / or Shore A hardness as disclosed herein. In a preferred embodiment, the frame portion and / or the microporous carrier portion are made from an elastomer as disclosed herein. To simplify the design and manufacture of the fluidic device of the present invention, the entire insert of a particularly preferred fluidic device is made from an elastomer as disclosed herein.

[0024] Similarly, the microporous carrier portion is preferably made from the same material as the frame portion, and more preferably the entire insert is made from the same material. In particularly preferred fluidic devices, the insert is a unitary piece. This means, among other things, that the microporous carrier portion and the frame portion are integral parts of the insert and are not, for example, glued together.

[0025] In a further preferred embodiment, the microporous carrier part and / or the frame part are characterized by a Shore A hardness of 80 or less and / or at least 2 (corresponding to a Shore OO hardness of 20). The Shore A hardness as disclosed herein defines the hardness of a particular part of the insert. Generally, the Shore A hardness can be determined on a standard specimen of the same material as the respective part to be evaluated. Different standard methods are available, such as DIN ISO 7619, DIN EN ISO 868, ASTM.D 2240 or JIS K 6253-3. For the purposes of the present invention, the Shore A hardness is preferably 80 or less and / or at least 2, measured according to DIN EN ISO 868. If the elastomer comprises further polymers and / or other components in addition to one polymer, the Shore A hardness as disclosed herein is meant to refer to the respective hardness resulting from the combination of all components present in the respective part.

[0026] According to this embodiment, the frame portion of the insert may have a hardness adapted to seal the structured area defining the flow path as disclosed herein against leakage of fluid. By using a frame portion having a hardness as disclosed herein, no additional sealing components such as gaskets are required. Instead, sealing can be obtained by compressing the layered structure, for example by placing the layered structure in a fluidic device holder as disclosed herein. With regard to the microporous carrier portion, a material having a hardness as disclosed herein is easily stretchable and provides sufficient elasticity to clamp the biological specimen securely. From the standpoint of simplicity of design and ease of manufacture, it is further preferred that the entire insert is characterized by a Shore A hardness of 80 or less and / or at least 2.

[0027] In a further preferred embodiment, the microporous carrier part is provided with an incision. As mentioned above, the incision is intended for clamping the biological specimen. The incision may be preformed or may be formed before use, for example by cutting with a knife, scissors, razor blade, etc. The microporous carrier part may also have a designated area for making the incision, which is optionally provided with a marking and / or a preformed break line (e.g., a notch), along which the microporous carrier part should be cut or break upon stretching. The incision or preformed break line disclosed herein may have a length of 0.3 to 30 mm, preferably 0.4 to 20 mm, more preferably 0.5 to 15 mm. It preferably does not extend into or through the frame part.

[0028] In a further preferred embodiment, the flow chamber and the at least one flow channel are formed by structuring the lower and / or upper surface of the insert, in which case the bottom plate and the cover plate can have a substantially flat shape and can be easily manufactured.

[0029] In a further preferred embodiment, the microporous carrier portion divides the flow chamber into a lower flow chamber region and an upper flow chamber region. Preferably, at least one lower flow channel is fluidly connected to the lower flow chamber region and at least one upper flow channel is fluidly connected to the upper flow chamber region. Such a configuration allows different fluids to be simultaneously supplied to the lower flow chamber region and the upper flow chamber region. For example, a first fluid containing nutrients can be supplied to the lower flow chamber region and a second fluid containing a staining agent can be supplied to the upper flow chamber region, or vice versa.

[0030] In a further preferred embodiment, the bottom plate and / or the cover plate comprises at least one port fluidly connected to at least one flow path. The supply of fluid through the ports contained in the cover plate is easily implemented and can be easily used with existing equipment (e.g., fluidic device holders).

[0031] To facilitate optical analysis, at least one of the bottom plate and the cover plate is preferably UV-VIS transparent. To this end, the bottom plate and / or the cover plate are preferably made of glass, such as borosilicate glass. In addition to being optically transparent, glass has the advantage of being hydrophilic, chemically inert, stable, and non-porous. However, it will be appreciated that it is also possible to utilize UV-VIS transparent plastics, such as PMMA or PS, depending on the wavelengths intended to be used for the analysis.

[0032] According to a second aspect of the present invention, the above problem is solved by an insert as disclosed herein. The insert includes, inter alia, a microporous carrier portion and a frame portion surrounding the microporous carrier portion, the insert being elastomeric as disclosed herein. Preferably, the frame portion and preferably also the microporous carrier portion are characterized by a Shore A hardness as disclosed herein. As mentioned above, the insert can include a plurality of microporous carrier portions, each microporous carrier portion being surrounded by a frame portion. The plurality of microporous carrier portions may be arranged in a rectangular pattern, for example including 4-20×6-40 carrier portions. The microporous carrier portions are preferably formed by recesses in the upper and / or lower surface of the insert. It is further preferred that at least one groove extends from each microporous carrier portion to provide at least one channel fluidly connected to each flow chamber as disclosed herein.

[0033] According to a third aspect of the present invention, the above problem is solved by a kit comprising an insert as disclosed herein, a cover plate as disclosed herein, and a bottom plate as disclosed herein. It should be understood that the bottom plate, the insert and the cover plate are such that when stacked in that order, they form a fluidic device as disclosed herein.

[0034] According to a fourth aspect of the present invention, the above problem is solved by a system comprising a fluidic device holder and a fluidic device as disclosed herein, an insert as disclosed herein, or a kit as disclosed herein. The fluidic device holder is understood to be adapted to engage with a fluidic device. To this end, the fluidic device holder comprises a base plate on which a first outer surface of the fluidic device (in particular the lower surface of the bottom plate as disclosed herein) is to be located, and a compression plate adapted to exert a compressive force on a second outer surface (in particular the upper surface of the top plate as disclosed herein) located opposite the first outer surface of the fluidic device. The compression plate is intended to provide a compressive force such that the fluidic device is clamped between the base plate and the compression plate of the fluidic device holder. Suitable device holders are commercially available, for example, from Micronit BV, Enschede, or Fluigent, Le Kremlin Bicetre, and are disclosed, for example, in PCT applications WO 2017 / 096296 A1 or WO 2009 / 002152 A1.

[0035] According to a fifth aspect of the present invention, the above problem is solved by a method for clamping a biological specimen, preferably a biopsy such as a prostate biopsy, the method comprising the steps of providing an insert as disclosed herein, wherein an incision is provided in a microporous carrier portion of the insert, and clamping the biological specimen between adjacent cut ends defined by the incision. As explained with respect to the function of the fluidic device disclosed herein, clamping the biological specimen between adjacent cut ends defined by the incision may be performed by exerting a stretching force (i.e. a pushing force, a pulling force or a bending force) such that an opening is formed between adjacent cut ends defined by the incision, the biological specimen is placed within the opening, the force is stopped, and thereby clamping the biological specimen between the adjacent cut ends. Preferably, the biological specimen is clamped such that a precise orientation of the biological specimen can be held, chemicals and nutrients can freely access the biological specimen from above and below, and / or a free optical view for microscopic examination can be provided.

[0036] Additionally, the methods disclosed herein preferably further comprise sandwiching an insert between a top plate and a cover plate to form a fluidic device disclosed herein.

[0037] Also preferred is a method as disclosed herein further comprising supplying at least one fluid to the flow chamber of the fluidic device, the fluid comprising at least one agent selected from the group consisting of nutrients, chemotherapeutic agents, staining agents, and agents for preparing a biopsy, in particular embedding and fixing agents such as paraffin and formalin. In one embodiment, the invention provides the use of a fluidic device as defined herein, or an insert as defined herein, or a system or kit as defined herein, in a procedure for containing a biological specimen, such specimen preferably being a biopsy specimen.

[0038] In one aspect, the invention provides a method of manufacturing an insert (7) for a microfluidic device (1), comprising forming an incision in an area of ​​a carrier portion of the insert (7) comprising a carrier portion (3) and a frame portion (8) surrounding the carrier portion (3), the insert (7) comprising or consisting of an elastomeric material, and the carrier portion comprising an area for forming the incision.

[0039] In some embodiments, methods of manufacturing a microfluidic device or a kit for a microfluidic device are provided, including methods of manufacturing an insert by assembling various parts of the kit into a kit, or by assembling parts of the kit into a device.

[0040] According to certain preferred methods disclosed herein, the biological specimen is examined pathologically and / or histologically, e.g., visually or microscopically. Prior to examination, the cover plate is removed in further preferred methods and the biological specimen, while still clamped within the insert, is cut into slices, e.g., using a microtome. In such methods, the insert is used as a holder for the microtome.

[0041] It is to be understood that the fluidic devices, inserts, kits, systems and methods disclosed in this specification, and in particular as disclosed in the accompanying independent claims, have similar and / or identical preferred embodiments, in particular as defined in the dependent claims.

[0042] It is to be understood that a preferred embodiment of the invention can also be any combination of the dependent claims or the various embodiments described herein with the respective independent claims.

[0043] These and other aspects of the invention will be apparent from and elucidated with reference to the embodiments described hereinafter. [Brief description of the drawings]

[0044] [Figure 1] FIG. 1 illustrates an example of a fluidic device disclosed herein. [Diagram 2] 13A-13C illustrate further examples of fluidic devices disclosed herein. [Diagram 3] FIG. 2 is an exploded bottom view of the fluidic device of FIG. 1 . [Figure 4] FIG. 2 is an exploded top view of the fluidic device of FIG. 1. [Diagram 5] FIG. 2 illustrates another example of a fluidic device disclosed herein. [Figure 6] FIG. 2 illustrates another example of a fluidic device disclosed herein. [Figure 7] FIG. 2 is a cross-sectional view of the fluidic device shown in FIG. [Figure 8A] FIG. 2 is a top view of a portion of a flow chamber having a microporous carrier portion. [Figure 8B] FIG. 2 is a top view of a portion of a flow chamber having a microporous carrier portion. [Figure 9] FIG. 13 illustrates an insert with multiple carrier portions. [Figure 10] Force-strain measurements obtained by pressing a round-tipped pen with a diameter of 1.2 mm against the opening of a membrane incision previously cut into the insert. [Figure 10A] FIG. 1 illustrates the measurement by showing the position of the pen at different times. [Figure 10B] Graph of force (F) versus strain (Δl). [Figure 11] 4 shows force strain measurements when an insert having cutouts as disclosed herein is stretched by pulling both ends of the insert in opposite directions. [Figure 11A] FIG. 1 illustrates the measurement by showing the apertures obtained at different times during the measurement. [Figure 11B] Graph showing force (F) versus strain (Δl). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0045] With reference to FIG. 1, the fluidic device 1 disclosed herein has a flow chamber 2 for accommodating a biological specimen on a microporous carrier portion 3 and at least one flow channel 4, 4a, 4b, 4c, 4d fluidly connected to the flow chamber 2. The fluidic device 1 has a layered structure comprising a bottom plate 5, a cover plate 6 and an insert 7 therebetween, as best seen in the exploded views of FIGS. 3 and 4. Furthermore, the insert 7 has a microporous carrier portion 3 and a frame portion 8 surrounding the microporous carrier portion 3, and is modified or modifiable with an incision 9 provided in the microporous carrier portion 3, as seen in FIG. 2. According to the present invention, the insert 7 is elastomeric so as to provide a clamping mechanism for the biological specimen as described herein.

[0046] To prevent damage to the biological specimen, the insert 7 is adapted so that the opening defined by the incision 9 can be widened using a small force to an extent sufficient to place the biological specimen therebetween. This is particularly useful when dealing with biopsies, which are generally very weak and sensitive to mechanical stress. The extent of widening and the force required depend on how the incision 9 has to be forced open.

[0047] When the opposing ends of the insert 7 are pulled apart in opposite directions, this force is preferably equal to or less than 5 N, more preferably equal to or less than 2.5 N, most preferably equal to or less than 1 N and / or at least 0.4 N, preferably at least 0.6 N. These values ​​apply in particular when the width of the opening defined by the incision 9 is widened to 0.2-3 mm, preferably 0.4-2 mm, more preferably 0.5-1.5 mm, most preferably 1 mm ±0.2 mm.

[0048] When a pushing force is applied in the thickness direction of the insert 7, for example using tweezers, directly or immediately adjacent to the incision 9, the force is preferably 50 mN or less, more preferably 10 mN or less, most preferably 5 mN or less and / or at least 0.5 mN, preferably at least 1 mN.

[0049] When a bending force is applied along an axis extending through the microporous carrier portion 3 such that the insert 7 is bent at least 60°, preferably at least 90°, more preferably at least 120°, even more preferably at least 150°, and most preferably about 180°, the force with which the insert 7 is gripped by the thumb and index finger is preferably 1000 mN or less, more preferably 500 mN or less, most preferably 200 mN or less, and / or at least 2 mN, preferably at least 5 mN. The flow chamber 2 disclosed herein represents a (micro)structured region within the fluidic device 1, having a volume sufficient to accommodate a biological specimen 10, preferably a biopsy such as a prostate biopsy, and at least one inlet and / or at least one outlet that connects to at least one channel 4a, 4b, 4c, 4d. A preferred flow chamber 2 is 50-1000 mm 3 Volume, for example, 100-500mm 3 . In certain embodiments, the flow chamber 2 further provides a microenvironment that allows the biological specimen 10 to be cultured and / or retain functional activity. In this case, the microporous carrier portion 3 can be provided with a correspondingly functionalized surface. For example, the microporous carrier portion 3 may be functionalized with collagen gel or fibronectin. In this way, cells or tissues can be accommodated in an environment similar to the one in which they naturally occur, which is particularly useful when the fluidic device is used to culture biological specimens (cells or tissues) for organ replacement / transplantation (organs on a chip), to perform ex vivo chemotherapy on biological specimens, or to test pharmaceuticals on biological specimens. Suitable functionalizing agents and methods are disclosed, for example, in PCT application WO2019 / 015988, entitled "Cell Culture Materials," the disclosure of which is incorporated herein by reference in its entirety, in particular for the purpose of disclosing agents and methods for functionalizing inserts for fluidic devices.

[0050] According to the illustrated embodiment, the flow chamber 2 has an essentially circular shape when viewed from above. More precisely, the flow chamber 2 preferably has the shape of a flattened cylinder. The flow chamber 2 can have, for example, a diameter of 2 to 30 mm, preferably 3 to 20 mm, more preferably 4 to 15 mm. Other shapes, such as oval or rectangular, are also conceivable.

[0051] The microporous carrier portion 3 and frame portion 8 as disclosed herein are best seen in Figure 2. Here it can also be seen that at least one flow path 4a, 4b (inlet and outlet paths in the embodiment of Figure 2) branches off from the flow chamber 2 and extends into the frame portion 8. Preferably, the bottom plate 5 and cover plate 6 are relatively rigid, while the insert 7 is relatively soft, as will be further described herein. With continued reference to Figure 2, it can be seen that an incision 9 is preferably provided in the microporous carrier portion 3.

[0052] The term "one channel" as used herein refers to configurations providing a single channel for fluid inflow and outflow, and configurations providing a pair of channels, one for fluid inflow (inflow channel) and the other for fluid outflow (outflow channel). Such a configuration consisting of a pair of inflow and outflow channels 4a, 4b is seen in FIG. 3. A configuration including two pairs of channels, each pair including an inflow channel 4a, 4c and a separate outflow channel 4b, 4d, is best shown in FIG. 1. The fluidic device 1 disclosed herein may include one flow path, two flow paths, three flow paths, four flow paths, five flow paths, six flow paths, or more than six flow paths, as disclosed above.

[0053] Referring again to the exploded view of the fluidic device 1 shown in Figure 3, it can be seen that the frame portion 8 of the insert preferably defines the side walls of the chamber and is intended to space the bottom plate 5 and the cover plate 6. This means that the flow chamber 2 is bounded by the upper surface of the bottom plate 5, the lower surface of the cover plate 6 and the inner wall of the frame portion 8. In the illustrated embodiment, the frame portion 8 is constituted by an unstructured area of ​​the insert 7. However, it will be readily understood that the frame portion 8 can alternatively be formed by ridges that surround the structured areas that define the flow chamber 2 and at least one flow path 4 (collectively referred to herein as flow paths 2, 4).

[0054] As best shown in Figures 3 and 4, the flow chamber 2 and at least one flow channel 4a, 4b, 4c, 4d of the fluidic device 1 disclosed herein are preferably formed by structuring the insert 7. However, it is also envisaged that at least one of the bottom plate 5 and the cover plate 6 is structured. In a preferred fluidic device 1, the lower surface of the insert 7 (i.e., the surface facing the bottom plate 5) and the upper surface of the insert 7 (i.e., the surface facing the cover plate 6) are provided with suitable structures to form the flow channels 2, 4 as disclosed herein. More specifically, the lower surface of the insert 7 is preferably provided with at least one groove in the area of ​​the frame portion 8 to form at least one channel 4a, 4b, as shown in Figure 3. Additionally or alternatively, the upper surface of the insert 7 may be provided with at least one groove in the area of ​​the frame portion 8 to form at least one channel 4c, 4d, as shown in Figure 4. The cross-sectional shape of the at least one groove corresponding to the at least one channel 4 is not limited, examples of possible shapes include a concave shape, a U-shape, and a V-shape. The depth and / or width of the groove is preferably 10 to 2000 μm, more preferably 50 to 1500 μm, and most preferably 100 to 1000 μm.

[0055] It is further preferred to provide the insert 7 with a recess on its underside (see FIG. 3) and / or on its top side (see FIG. 4) in the region of the carrier part 3 in order to form the flow chamber 2. According to this configuration, the flow chamber 2 is formed by a microporous carrier part 3, the layer thickness of which is smaller than that of the surrounding frame part 8. Such a preferred insert 7 can be produced by any suitable technique for producing surface-structured, preferably elastomeric, moulded articles. Preferred techniques include, but are not limited to, moulding, 3D printing, casting and embossing.

[0056] With continued reference to Figures 3 and 4, it can be seen that the fluid is preferably supplied through the cover plate 6. To this end, the cover plate 6 comprises at least one port 11 providing fluid access to at least one channel 4a, 4b, 4c, 4d. Furthermore, it can be seen that the insert 7 can include at least one opening 12 aligned with the at least one port 11, the at least one opening 12 being fluidly connected to the flow chamber 2 via the at least one channel 4. In the embodiment shown in Figures 3 and 4, the at least one port 11 and the at least one opening 12 have a circular shape with coincident radial axes. As mentioned above, the supply of fluid through the cover plate 6 can be easily implemented and easily used with existing equipment (e.g. fluidic device holders), but it is also envisaged that the fluid can be supplied through the bottom plate 5 or through the side of the insert 7, in which case a corresponding port 11 is present in the bottom plate 5 or in the insert 7.

[0057] As shown in Figures 5 and 6, the ports 11 may be connected to fittings 13 that can be connected to tubes carrying fluids. The embodiment shown in Figure 5 has a configuration with four fittings 13 that provide fluid access to the lower flow channel (not visible in Figure 5) and the upper flow channels 4c, 4d as disclosed herein. It can further be seen that the cover plate 6 is provided with a number of ports 11, while the number of functional ports (i.e. ports that provide fluid access to the flow chamber 2 via at least one of the flow channels 4a, 4b, 4c, 4d) is governed by the structured surface of the insert 7. In this example, the fluidic device 1 comprises four functional ports and eight dead-end ports 11a that do not align with corresponding openings 12 in the insert 7 and are therefore non-functional. In the configuration shown in Figure 6, all twelve ports are provided with fittings 13 and are functional. The structure of the insert 7 is not shown. This embodiment has the advantage that an equally configured cover plate 6 can be used regardless of the desired number of functional ports.

[0058] The insert 7 is preferably a flat rectangular parallelepiped with upper and lower surfaces facing the lower surface of the cover plate 6 and the upper surface of the bottom plate 5, respectively. The upper surface of the flat rectangular parallelepiped and / or the lower surface of the structured area of ​​the flat rectangular parallelepiped are provided with the flow channels 2, 4 as disclosed herein. More preferably, the upper and lower surfaces of the insert 7 are rectangular with long and short sides, and the cutouts are provided substantially parallel to the length of the short side (at ±45°, preferably ±30°, more preferably ±20°). Other shapes such as a flat cuboid shape with rounded edges or a flat cylinder with corresponding structures as previously described are equally conceivable. More preferably, the shape of the insert 7 is such that its upper and lower surfaces correspond to the lower surface of the cover plate 6 and the upper surface of the bottom plate 5, respectively.

[0059] The layer thickness of the insert 7, which is preferably a flat rectangular parallelepiped, measured at its thickest point may be in the range of 0.5 to 20 mm, preferably 1 to 15 mm, more preferably 2 to 10 mm, and most preferably 3 to 8 mm. The layer thickness of the insert 7, measured at its thinnest point, may be in the range of 0.1 to 5 mm, preferably 0.1 to 3 mm, more preferably 0.2 to 2 mm, and most preferably 0.3 to 1 mm. In a preferred fluidic device 1, the thickest point is in the frame part 8 and the thinnest point is in the carrier part 3. In this case, the thickness of the carrier part 3 may be, for example, 5 to 50% of the thickness of the frame part 8.

[0060] A preferred fluidic device 1 disclosed herein comprises a flat bottom plate 5 and / or a flat cover plate 6 having a flat cuboid shape, optionally with rounded edges or a flat cylindrical shape, as described above with respect to the insert 7. Preferably, the thickness of the flat cuboid bottom plate 5 and / or cover plate 6 may be in the range of 0.5-20 mm, preferably 0.8-15 mm, more preferably 1-10 mm, and most preferably 2-6 mm.

[0061] Referring now to FIG. 7, a cross-sectional view of the fluidic device 1 shown in FIGS. 3 and 4 is shown. As can be seen, in the preferred fluidic device 1, a carrier portion 3 separates the flow chamber 2 in a lower flow chamber region 2a and an upper flow chamber region 2b. In this case, at least one lower flow channel 4a, 4b is provided in fluid communication with the lower flow chamber region 2a, and more preferably at least one upper flow channel 4c, 4d is provided in fluid communication with the upper flow chamber region 2b. With such a configuration, the lower flow chamber region 2a and the upper flow chamber region 2b can be simultaneously supplied with different fluids, as described herein. Also, as described herein, the at least one channel 4 is preferably a pair of channels, the pair providing inlet channels 4a, 4c and outlet channels 4b, 4d.

[0062] The structure of the microporous carrier part 3 of the preferred fluidic device 1 will now be described with reference to FIG. 8. This structure is in particular such that a cross-sectional fluid flow through the insert occurs, thereby allowing constant and reproducible perfusion, and that the insert 7 can be reversibly bent along an axis extending through the microporous carrier part 3. The number and size of the pores as well as the pore density can be selected as required. Furthermore, the pores can be randomly distributed or arranged in a geometrically regular pattern. In the microporous carrier part 3 shown in FIG. 8(a), the pores are arranged in a geometrically regular and symmetrical pattern. Such a pattern can be polygonal, for example octagonal as shown, and can have any other form including rectangular, circular or elliptical. According to exemplary embodiments, the microporous carrier part 3 as disclosed herein can include a large number of pores, for example 8-200 pores, 12-150 pores, 16-100 pores or 20-80 pores. In this case, the average diameter of the pores is preferably relatively large, for example, 100 μm to 1000 μm, 200 μm to 750 μm, or 250 μm to 500 μm. The pore density is 8 to 200 pores / cm. 2 , 12-150 pores / cm 2 , 16-100 pores / cm 2 , 20-80 pores / cm 2 As shown in FIG. 8B, the microporous carrier part 3 according to another exemplary embodiment includes 20-1500 pores, 50-1000 pores, or 100-750 pores. In this case, the average diameter of the pores is relatively small, for example, preferably 5 μm-100 μm, 8 μm-75 μm, or 10 μm-50 μm. The pore density is preferably 20-1500 pores / cm. 2 , 50-1000 pores / cm 2 , or 100 to 750 pores / cm 2 The pores can be conveniently produced by molding with a suitable matrix or by forming a mesh using conventional weaving techniques.

[0063] With reference to FIG. 9, it can be seen that the fluidic device 1 disclosed herein is not limited to having only one flow chamber 2. Rather, the fluidic device 1 disclosed herein may have two or more flow chambers 2. For example, there may be multiple flow chambers and multiple corresponding carrier portions 3, 3', 3" may be formed in an insert 7 sandwiched between a bottom plate 5 and a cover plate 6 as disclosed herein. In the embodiment shown in FIG. 9, the flow chambers 2 are arranged in a pattern including 6 columns by 4 rows. A single bottom plate 5 and a single cover plate 6 extend on opposing surfaces of the insert 7, thus sealing the multiple flow chambers 2a, 2b, 2c and the flow channels 4. The cover plate 6 further comprises multiple ports 11 that provide fluid access to the multiple flow chambers 2a, 2b, 2c via the flow channels 4.

[0064] Referring to Figures 10 and 11, the results of a stress strain measurement on an insert 7 as disclosed herein are shown. Figure 10 is a measurement simulating the force required to push tweezers with a biological sample between the incisions 9, leaving the biological sample behind. The force required to manipulate the opening and closing of the insert 7 was simulated on a Zwick Z010 stress bench by pushing a pen with a blunt tip and a thickness-wise diameter of 1.2 mm over and through the incisions 9 pre-cut in the insert 7. After three individual measurements, this force was determined to be on the order of 3 mN.

[0065] FIG. 11 is a simulated measurement of the force required to widen, by pulling, the opening defined by the incision 9 to such an extent that the widened opening can receive a biological specimen, for example to a width of 1 mm. The force required to manipulate the opening of the insert 7 was simulated on a Zwick Z010 stress bench by pulling both ends of the insert 7 in opposite directions. The insert 7 was made from silicone (Wacker Elastosil 3040 / 40) with a Shore A hardness of 40. The microporous carrier part 3 of the insert 7 had a thickness of 35 μm and a pore size of 200 μm. According to an exemplary measurement, the results of which are shown in FIG. 11, it was determined that the force to widen the opening to a width of 1 mm was of the order of 800 mN.

[0066] Other variations to the disclosed embodiments can be understood and effected by those skilled in the art in practicing the claimed invention, from a study of the disclosure and the appended claims.

[0067] In the claims, the word "comprise" does not exclude other elements or steps and the indefinite article "a" or "an" does not exclude a plurality.

[0068] A single unit or device may fulfill the functions of several items recited in the claims. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage.

[0069] Any reference signs in the claims should not be construed as limiting the scope.

Claims

1. 1. A kit including parts for manufacturing a fluidic device having a flow chamber for containing a biological specimen in a carrier portion and at least one flow path fluidly connected to the flow chamber, The components include a bottom plate, a cover plate, and an insert; the insert has a carrier portion and a frame portion surrounding the carrier portion, the carrier portion having or consisting of an elastomeric material, the carrier portion having an incision for clamping the biological specimen or a region having an indicator for guiding a user in forming the incision for clamping the biological specimen; when the components are assembled into the fluidic device, the insert is sandwiched between the bottom plate and the cover plate such that the flow chamber is defined by an upper surface of the bottom plate, a lower surface of the cover plate and an inner wall of the frame portion of the insert. kit.

2. The kit of claim 1 , wherein the frame portion comprises or consists of an additional elastomeric material.

3. 3. The kit according to claim 1 or 2, wherein the carrier part and / or the frame part are characterized by a Shore A hardness ranging from 2 to 80.

4. The kit of claim 1 , wherein the carrier portion has an incision that defines an opening in the carrier portion when the carrier portion is in an undeformed state.

5. 5. The kit according to claim 1, wherein the elastomeric material and / or the further elastomeric material is selected from the group consisting of thermoplastic elastomers and elastomers such as silicone and polybutadiene rubber, the thermoplastic elastomer being selected from the group consisting of styrene block copolymers (TPS), styrene ethylene butylene styrene block copolymers (SEBS) and thermoplastic polyurethanes (TPU).

6. The kit of claim 1 , wherein the carrier portion is porous or microporous.

7. The kit of claim 1 , wherein the lower and / or upper surface of the insert has a groove for forming the at least one flow channel.

8. 8. The kit of claim 1, wherein the carrier portion divides the flow chamber into a lower flow chamber region and an upper flow chamber region, at least one lower flow channel being fluidly connected to the lower flow chamber region and at least one upper flow channel being fluidly connected to the upper flow chamber region.

9. 9. The kit of claim 1, wherein the bottom plate and / or the cover plate have at least one port that is fluidly connected to the at least one flow path when the parts are assembled into the fluidic device.

10. 10. The kit of claim 1, wherein the bottom plate and / or the cover plate are UV-Vis transparent.

11. the bottom plate and / or the cover plate have a layer thickness of 0.5 to 20 mm, and / or the insert has a layer thickness, measured at its thickest point, of 0.5 to 20 mm; and / or 11. The kit according to any one of claims 1 to 10, wherein the insert has a layer thickness, measured at its thinnest point, of 0.1 to 5 mm.

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