Holding device, in particular tissue holding device and method for receiving a tissue sample

Abstract

A holding device (10), in particular a tissue holding device, for holding and positioning a tissue sample, in particular a tissue section, is described, with a tissue receiving area (3), wherein the tissue receiving area (3) has a receiving platform (3a) with a plurality of tissue-fixing structures (5) which are designed to at least partially penetrate the tissue sample and to stretch it over a surface.

Description

[0001] The present invention relates to a holding device, in particular a tissue holding device for holding and positioning a tissue sample over a flat area, to a method for manufacturing the same, and to a method for receiving a tissue sample by means of the same according to the preamble of the independent claims. State of the art

[0002] In modern cancer medicine, the goal is to tailor therapy as closely as possible to the needs of each individual patient. Efforts are made to address individual circumstances (personal characteristics and genetic predispositions). Based on molecular genetic analyses of, for example, tumor tissue, the aim is to target the pathologically altered cells as effectively as possible and, if possible, to avoid non-specific chemotherapy, which attacks all rapidly growing cells and with all its associated side effects. This approach seeks to protect healthy body cells as much as possible and to prevent the growth and spread of diseased tumor cells in a targeted manner.

[0003] For this purpose, it is necessary to cultivate tissue, especially tumor tissue, in order to test the effects of drugs on it ex vivo. This is possible, for example, using fluorescence microscopes with a cultivation unit, in which samples are cultured, stained, and treated with active substances to test their efficacy.

[0004] To obtain good optical access to biopsied tissue, the punched or otherwise obtained samples are cut into thin slices and placed in the culture medium. These tissue sections have, for example, dimensions of 5 mm laterally and a thickness of 200 µm. When placed in the receiving containers, they curl up undefinedly, clump together, or move uncontrollably when the culture vessels are moved, thus severely impairing the introduction of light and optical accessibility.

[0005] US patent 201070208049 A1 discloses a device for imaging a tissue sample. The tissue sample is held in a culture chamber. To fix the tissue sample in the culture chamber, it is clamped between a window and a platform. The platform includes microneedles, which, however, do not serve a holding function but rather inject fluid into the tissue sample.

[0006] DE 102019101035 A1 discloses a sample holder for a tissue sample with two plates that are held together by means of magnetic elements and between which a tissue sample can be clamped. Disclosure of the invention

[0007] Against this background, a holding device, in particular a tissue holding device, a method for manufacturing the same, and a method for holding and positioning a tissue sample over a surface with the characterizing features of the independent claims are provided according to the invention.

[0008] The holding device according to the invention, in particular a tissue holding device, is designed for holding and positioning a tissue sample, in particular a tissue section, over a flat surface.

[0009] A tissue sample can refer, for example, to a biopsied tissue sample from plants, animals or humans, with a thickness between 10µm and 10mm.

[0010] In the context of the present invention, "flat holding" means that the tissue sample, for example the tissue section, is held stretched across a flat surface and cannot curl up or clump together.

[0011] The holding device includes a tissue receiving area. This area comprises a receiving platform, which can be round or square, ring-shaped or forked. A number of tissue-fixing structures are arranged on the receiving platform, by means of which a tissue sample can be at least partially penetrated, thereby held, and stretched across its surface.

[0012] The advantage here is that the tissue sample retains its flat shape when held by the holding device, thus ensuring optimal accessibility. This is particularly beneficial for microscopic, and especially fluorescence microscopy, examination of the tissue sample, as optical accessibility and the introduction of light for examination are crucial factors. Furthermore, the device prevents a tissue sample held by the holding device from moving uncontrollably in the sample liquid when the sample or culture vessel into which the holding device can be inserted is moved, which would otherwise impair optical examination.

[0013] Furthermore, the holding device prevents a tissue sample in a sample or culture vessel from moving uncontrollably in the sample fluid when the vessel is moved, which could, for example, impair optical examination. For this purpose, the holding device can be inserted into the sample or culture vessel.

[0014] The holding device immobilizes the tissue sample in such a way as to ensure optimal conditions for its examination, particularly optical examination. This also makes the tissue sample easily accessible for culture media, test substances, drugs, intercalating dyes, antibodies, and the like.

[0015] Furthermore, the holding device can be advantageously used for transporting the tissue sample. For this purpose, the tissue sample is first picked up by the holding device and then immersed in liquid nitrogen, so that the tissue sample can be transported frozen in a flat shape, either clamped in the device and together with it, or alternatively without it.

[0016] Further advantageous embodiments of the present invention are the subject of the dependent claims.

[0017] It is advantageous if the holding device is made of a metal, in particular a corrosion-resistant steel and / or a corrosion-resistant stainless steel and / or a corrosion-resistant spring steel, and / or if the holding device includes titanium.

[0018] Advantages of using corrosion-free spring steel include its corrosion resistance, long service life, easy formability, and high strength. Furthermore, the strength properties can be specifically adjusted and thus adapted to the application.

[0019] The advantages of using titanium include its high strength, toughness, and hardness combined with low density, making it both stable and lightweight. Titanium is also corrosion- and temperature-resistant and biocompatible. Alternatively or additionally, it is advantageous if the holding device comprises a thermoplastic material, in particular polypropylene (PP), polyethylene (PE), polyethylene terephthalate (PET), polyetheretherketone (PEK), polytetrafluoroethylene (PTFE), Teflon, and / or polytetrafluoroethylene (TPFE).

[0020] Compared to metallic materials, their lighter weight is an advantage.

[0021] Furthermore, a holding device made of a thermoplastic material is cost-effective.

[0022] A further advantage of all the aforementioned materials is their biocompatibility and the fact that they are non-cytotoxic, meaning the tissue sample is not affected by the holding device. These materials can also be sterilized or autoclaved, ensuring the holding device is sterile for use and reusable, thus allowing for multiple uses.

[0023] In an advantageous embodiment, the tissue-fixing structures of the tissue receiving area are pointed, in particular pin-like, needle-shaped, pyramidal, and / or conical structures. These are arranged, for example, at an angle γ to the main axis of the receiving platform. The angle γ is, for example, 70°–110° and advantageously 80°–100°. The tissue-fixing structures can be bent upwards or downwards with respect to the receiving platform, or located above or below it, so that the tissue can be received and held above or below the receiving platform.

[0024] The tissue-fixing structures, for example, have a length of less than 1 mm. Pin-like, needle-shaped, pyramid-shaped and / or cone-shaped tissue-fixing structures can be manufactured, for example, by injection molding or other molding processes.

[0025] The advantage of such tissue-fixing structures is that they can penetrate the tissue sample easily and without much resistance, without causing significant damage.

[0026] If the tissue-fixing structures are arranged at an angle γ on the receiving platform, so that they point outwards, i.e. away from the center of the receiving platform, the optically accessible area of ​​the tissue sample is larger.

[0027] In an advantageous embodiment, the tissue-fixing structures are arranged orthogonally on the receiving platform and are slightly bent when the platform is pressed into the tissue against a stable surface or by a snap-action mechanism, so that the tissue-fixing structures point away from the center of the receiving platform and the tissue sample is advantageously tautened when the tissue-fixing structures penetrate and / or when the tissue sample is lifted.

[0028] Alternatively or additionally, the tissue-fixing structures of the tissue receiving area have barbs and are, for example, arrow- or hook-shaped. The advantage here is that a tissue sample can be easily, at least partially, penetrated, and the tissue sample adheres or hooks onto the barbs, thus achieving a good holding effect and preventing the tissue sample from slipping off the tissue-fixing structures.

[0029] The tissue-fixing structures with barbs are advantageously arranged at an angle to the main axis of the receiving platform and, for example, have a length of less than 1 mm. If the tissue-fixing structures with barbs are arranged at an angle γ on the receiving platform, so that they point outwards, i.e., away from the center of the receiving platform, the advantages mentioned above result.

[0030] Alternatively or additionally, the fabric-fixing structures are formed by hook-shaped geometries similar to the hook strips of Velcro fasteners. Advantageously, the receiving platform has at least one initial recess, which is round, oval, and / or angular in shape. The fabric-fixing structures are arranged along the edge of this initial recess. The receiving platform can also have several such recesses.

[0031] An advantage of having one or more recesses in the receiving platform is that the tissue sample is held firmly and securely and cannot fold, slip out or curl up on one side, thus ensuring a flat holding and positioning of the tissue sample held by the receiving platform.

[0032] In a further advantageous embodiment, the receiving platform is designed in a fork shape and the tissue-fixing structures are arranged at least along the tines of the fork, or the receiving platform is designed in a grid shape and the tissue-fixing structures are arranged along the grid.

[0033] A benefit of gripping jaws with a grid-like design is that even small tissue samples can be held very securely.

[0034] An advantage of a fork-shaped receiving platform is that it allows good access to the tissue sample and thus good optical analysis of the tissue sample.

[0035] A grid-shaped receiving platform offers the advantage that even small tissue samples can be held very securely.

[0036] In a particularly advantageous embodiment, the holding device has a retaining leg. The advantage here is that this allows for good and simple handling of the holding device. Furthermore, it is particularly advantageous if the tissue receiving area is arranged at an angle α of 45°–175°, and preferably 125°, on the retaining leg.

[0037] Depending on the circumstances and conditions, this angle can easily be changed.

[0038] Furthermore, a holding device made of hybrid materials is also conceivable. For example, the holding arm is made of a metal, particularly corrosion-resistant stainless steel, and the tissue receiving area and / or the tissue-fixing structures comprise a thermoplastic material, or vice versa. The tissue receiving area can be a disposable component. This can, for example, be attached to the holding arm and / or snapped into place. Additionally, metallic tissue-fixing structures can have a coating, for example, of silicone, heparin, and / or hyaluronic acid. This can, for example, prevent rejection reactions of the tissue sample.

[0039] In a further advantageous embodiment, the holding device is bent in such a way that it has at least three predefined contact areas which, when the holding device is inserted into a sample container, in particular a cultivation vessel, bear against its inner walls. The shape of the holding device is, for example, adapted to the shape of the sample container, in particular the cultivation vessel.

[0040] The advantage here is that the holding device is statically pre-positioned in the sample container in this way and cannot, for example, slip, so that a particularly good optical examination of the tissue sample in the sample container can be carried out.

[0041] Furthermore, it is advantageous if the holding device has second recesses at the bending points of the holding leg.

[0042] The advantage here is that the holding device can be bent easily and precisely at the bending points.

[0043] Another object of the invention relates to a method for manufacturing the holding device according to the invention, in particular a tissue holding device, for holding and positioning a tissue sample over a flat surface, comprising the following steps: a) Provision of a metal sheet, in particular made of corrosion-resistant steel and / or corrosion-resistant stainless steel and / or corrosion-resistant spring steel and / or titanium b) Cutting out, in particular by laser cutting, milling, punching and / or etching, the metal sheet so that a one-piece, two-dimensionally contoured metal sheet is obtained. Optionally, an initial recess for the receiving platform can be created, for example by means of a hole. c) Multiple bending of the metal sheet. Here, the retaining leg is bent in a defined manner at the bending points, and the pointed, in particular arrow- or hook-shaped, fabric-fixing structures are bent at an angle γ to the main axis of the receiving platform. The fabric-fixing structures can be bent upwards or downwards relative to the receiving platform, so that the fabric can be picked up above or below the receiving platform.

[0044] Optionally, in a further step d) a heat treatment, in particular a stress-relieving annealing, can be carried out to improve the material structure and to reduce or eliminate any residual stress unintentionally introduced by bending (cold forming) and to prevent unwanted distortion.

[0045] Alternatively, the holding device according to the invention is manufactured using an injection molding process with the following steps: a') Providing a thermoplastic material, in particular PP, PE, PET, PEK, PTFE and / or TPFE for an injection molding process b') Injection molding of the holding device

[0046] The tissue-fixing structures are pointed, in particular pin-like, needle-shaped, pyramid-shaped and / or cone-shaped structures.

[0047] Alternatively, the holding device according to the invention is manufactured using an additive process with the following steps: a'') Providing a metal and / or a plastic b'') Additive manufacturing of the holding device

[0048] Each of the aforementioned methods for manufacturing the holding device yields one such device comprising a holding leg and a tissue receiving area with a receiving platform having a plurality of tissue-fixing structures.

[0049] The invention further relates to a method for taking a tissue sample, in particular a tissue section, using the holding device comprising the following steps: e) Providing a holding device according to the invention, in particular a tissue holding device f) Positioning the holding device over the tissue sample to be received g) Applying a compressive force to the holding leg and / or the back of the tissue receiving area, so that the tissue-fixing structures of the receiving platform at least partially penetrate the tissue sample and hold it stretched flat. h) optional positioning of the holding device with the collected tissue sample in a sample container, in particular in a culture vessel.

[0050] The advantages resulting from the method for taking a tissue sample correspond to those mentioned for the device.

[0051] The holding device according to the invention is advantageously used, for example, for clamping a tissue sample, in particular a tissue section, over a flat surface, and for positioning it, in particular, in a sample container, for example, a cultivation vessel of a microscope, such as a fluorescence microscope. In the cultivation vessel, the tissue sample can, for example, be cultured, stained, and treated with active substances to test their efficacy.

[0052] Furthermore, the holding device can also be used to transport a tissue sample clamped into it, so that the tissue sample is already available in a flat, stretched form at the place of use and can be inserted directly into a sample container, for example a cultivation vessel, using the holding device. Brief description of the drawing

[0053] The drawing illustrates advantageous embodiments of the present invention, which are explained in more detail in the following description. It shows: Fig. 1: A schematic 3D representation of a holding device according to a first embodiment of the present invention, Fig. 2a: an enlarged schematic 3D representation of the tissue receiving area of ​​the holding device according to the invention. Fig. 1 in a first embodiment, Fig. 2b: a schematic 3D representation of the tissue holding receiving area of ​​the holding device according to the invention in a second embodiment, Fig. 2c: a schematic 3D representation of the tissue holding receiving area of ​​the holding device according to the invention in a third embodiment, Fig. 3a: a schematic 3D frontal view of the holding device according to the invention Fig. 1 with marked first investment areas, Fig. 3b: a schematic side view of the holding device according to the invention Fig. 1 and Fig. 3a with marked second planting areas, Fig. 4: The schematic representation of a flowchart of an embodiment of the inventive method for holding a tissue sample.

[0054] In Fig. Figure 1 shows a first embodiment of a holding device 10 according to the invention. The holding device 10 comprises a holding leg 1 and a tissue receiving area 3. The holding leg 1 is in Fig. The retaining leg 1 is divided into a first retaining leg region 1a, a second retaining leg region 1b, and a third retaining leg region 1c, between each of which lies a bending point 8 at which the retaining leg 1 is bent. The tissue receiving area 3 adjoins the third retaining leg region 1c via a further bending point 8. This area is arranged at an angle α of 45°–175°, and preferably 125°, on the retaining leg 1.

[0055] Alternatively and not in Fig. As shown in Figure 1, the retaining leg 1 has more or fewer bending points 8. Fig. 1. The retaining leg 1 has a second recess 9 at each of the bending points 8. Further alternative and not in Fig. As shown in Figure 1, the retaining leg 1 has no second recesses 9. Alternatively, the retaining leg 1 has a perforation or a local reduction in material thickness without penetration at the bending points 8. The aforementioned measures, which facilitate the bending process, can also be combined.

[0056] The tissue receiving area 3 comprises a receiving platform 3a, on which a plurality of tissue-fixing structures 5 are arranged. By means of the tissue-fixing structures 5, a tissue sample can be at least partially penetrated, so that it is held stretched flat.

[0057] The holding device 10 is made, for example, of corrosion-free steel and / or corrosion-free stainless steel and / or titanium and / or a thermoplastic material, such as PP, PE, PET, PEK, PTFE and / or TPFE.

[0058] In Fig. 2a is the tissue receiving area 3 of the holding device 10 according to Fig. Figure 1 shows an enlarged view rotated 180°. The receiving platform 3a has a square shape with rounded corners. In the center, the receiving platform 3a has a circular first recess 7. Along the edge of the first recess 7, tissue-fixing structures 5 are arranged at an angle to it on the receiving platform 3a. The tissue-fixing structures 5 are arrow-shaped. A tissue sample can be at least partially penetrated by means of the arrowhead, while the lateral corners of the arrow structures act as barbs in the tissue and ensure a firm hold. The length of the tissue-fixing structures 5 is, for example, less than 1 mm.

[0059] In Fig. Figure 2b shows the tissue receiving area 3 of the holding device 10 according to the invention in a second embodiment. In contrast to Fig. 2a The tissue-fixing structures 5 are designed in a hook shape. A tissue sample can be at least partially penetrated by means of the highest point of the hook-shaped structures 5, while the laterally arranged, pointed, protruding hook of each tissue-fixing structure 5 serves as a barb and hooks into the tissue.

[0060] In Fig. Figure 2c shows the tissue receiving area 3 of the holding device 10 according to the invention in a third embodiment. The receiving platform 3a has a square shape with unrounded corners. In the center, the receiving platform 3a has a first recess 7, also square. Tissue-fixing structures 5 are arranged on the receiving platform 3a at a 90° angle to the edge of the first recess 7. The tissue-fixing structures 5 are pyramid-shaped. A tissue sample can be at least partially penetrated by means of the apex of the pyramid, and the tissue is ultimately held by the pyramid shape of the tissue-fixing structures 5. The length of the tissue-fixing structures 5 is, for example, less than 1 mm.

[0061] Alternatively, and not shown in the figures, the first recess 7 can also have an oval or differently angular shape. Likewise, the receiving platform 3a can have a round, polygonal, or any other shape with rounded or pointed corners. The tissue-fixing structures 5 can be bent upwards or downwards with respect to the main axis of the receiving platform 3a, or attached, for example, by injection molding.

[0062] Alternatively, and not shown in the figures, the receiving platform 3a is, for example, designed in a fork shape and the tissue-fixing structures 5 are arranged at least along the tines of the fork, or the receiving platform 3a is designed in a grid shape and the tissue-fixing structures 5 are arranged along the grid structure.

[0063] Furthermore, the tissue-fixing structures can alternatively or additionally be pin-like, needle-shaped, pyramid-shaped and / or cone-shaped structures.

[0064] In Fig. 3a is a frontal view of the holding device 10 according to Fig. Figure 1 shows the first contact areas 11a on the lateral right and left edges of the second retaining leg 1b, as well as first contact areas 11a on the lateral right and left edges of the receiving platform 3a. In this embodiment, the holding device 10 is specifically designed with respect to a defined sample container, in particular a cultivation vessel, such that when the holding device 10 is inserted into the corresponding sample container, the first contact areas 11a at least touch or abut its side walls. The holding device 10 is then not movable to the left or right.

[0065] In Fig. 3b is a side view of the holding device 10 according to the Fig. 1 and Fig. 3a with additionally marked second contact areas 11b shown. The front main surface of the first retaining leg area 1a forms a second contact area 11b, the rear main surface of the third retaining leg area 1c forms a second contact area 11b and the lower edge of the receiving platform 3a also forms a second contact area 11b (in Fig. 3b (shown only as a dot). Here too, the holding device 10 is specifically designed with regard to a defined sample container, in particular a cultivation vessel, so that when the holding device 10 is inserted into the corresponding sample container, the second contact areas 11b at least touch or rest against its side walls. The holding device 10 is then not movable forwards or backwards. The arrow in the illustration of the second contact areas 11b is intended to provide a clear overview of the second contact areas 11b.

[0066] The first 11a and the second installation areas 11b can be combined so that the holding device 10 is defined and can be firmly inserted into the sample container and cannot move through the first and second installation areas.

[0067] Fig.Figure 4 shows an embodiment of a method 100 for taking up a tissue sample, in particular a tissue section, using the holding device 10 according to the invention.

[0068] After the holding device 10 is provided in step k), in step I) it is positioned over a tissue sample to be received, in particular a tissue section. Subsequently, in step m), a pressure force is applied to the holding arm 1, so that this force is transferred to the tissue receiving area 3 and the tissue-fixing structures 5 of the receiving platform 3a at least partially penetrate the tissue sample and hold it firmly in place. If the tissue-fixing structures 5 have barbs, these engage in the tissue and hold it securely.

[0069] In an optional step n), the clamped tissue sample is positioned in a sample container, in particular in a culture vessel 17. QUOTES INCLUDED IN THE DESCRIPTION

[0000] This list of documents cited by the applicant was automatically generated and is included solely for the reader's convenience. The list is not part of the German patent or utility model application. The DPMA accepts no liability for any errors or omissions. Cited patent literature

[0000] US 201070208049 A1

[0005] DE 102019101035 A1

[0006]

Claims

[1] Holding device (10), in particular tissue holding device, for holding and positioning a tissue sample, in particular a tissue section, with a tissue receiving area (3), wherein the tissue receiving area (3) has a receiving platform (3a) with a plurality of tissue-fixing structures (5) which are designed to at least partially penetrate the tissue sample and to stretch it over a surface. [2] Holding device (10) according to claim 1, wherein the holding device (10) comprises a metal, in particular a corrosion-resistant steel and / or a corrosion-resistant stainless steel and / or titanium and / or a thermoplastic polymer, in particular polypropylene (PP), polyethylene (PE), polyethylene terephthalate (PET), polyetheretherketone (PEK), polytetrafluoroethylene (PTFE), Teflon and / or polytetrafluoroethylene (TPFE). [3] Holding device (10) according to one of the preceding claims, wherein the tissue-fixing structures (5) of the receiving platform (3a) have pointed, in particular pin-like, needle-shaped, pyramid-shaped and / or conical structures, in particular with a length of less than 1 mm, which are in particular arranged at an angle to the main axis of the receiving platform (3a). [4] Holding device (10) according to one of the preceding claims, wherein the tissue-fixing structures (5) of the receiving platform (3a) are structures with barbs, in particular arrow- or hook-shaped structures, in particular with a length of less than 1 mm, which are in particular arranged at an angle to the main axis of the receiving platform (3a). [5] Holding device (10) according to one of the preceding claims, wherein the receiving platform (3a) has at least one first recess (7) which has a round, oval and / or angular shape and wherein the tissue-fixing structures (5) are arranged along the edge of the first recess (7). [6] Holding device (10) according to one of the preceding claims, wherein the receiving platform (3a) is designed in a fork shape and the tissue-fixing structures (5) are arranged at least along the tines of the fork or wherein the receiving platform (3a) is designed in a grid shape and the tissue-fixing structures (5) are arranged along the grid. [7] Holding device (10) according to one of the preceding claims, wherein the holding device (10) has a holding leg (1), and wherein the tissue receiving area (3) is arranged at an angle α of 45°- 175°, and preferably of 125° on the holding leg (1). [8] Holding device (10) according to one of the preceding claims, wherein the holding device (10) is bent such that it has at least three predefined contact areas (11a, 11b) which, when the holding device (10) is inserted into a sample container, in particular a cultivation vessel, bear against its inner walls, and in particular that the holding device (10) has second recesses (9) at the bending points (8). [9] Method for manufacturing a holding device (10), in particular a tissue holding device, according to one of claims 1-8, for holding and positioning a tissue sample over a surface, comprising the steps: a) Provision of a metal sheet, in particular made of corrosion-resistant steel and / or corrosion-resistant stainless steel and / or titanium b) Cutting, punching and / or etching the metal sheet to obtain a one-piece, two-dimensionally contoured metal sheet c) Multiple bending of the metal sheet. [10] Method for taking up a tissue sample, in particular a tissue section, using the holding device (10), in particular tissue holding device, according to one of claims 1-8 comprising the steps: k) Providing a holding device (10) I) Positioning the holding device (10) over the tissue sample to be received m) Applying a pressure force, in particular to the retaining leg (1) and / or the back of the tissue receiving area (3), so that the tissue-fixing structures (5) of the receiving platform (3a) at least partially penetrate the tissue sample and hold it stretched across its surface n) optional positioning of the holding device (10) with the received tissue sample in a sample container, in particular in a culture vessel (17).

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