An apparatus for preparing a multi-sample single cell suspension slide microarray

Abstract

The application relates to the technical field of cytological detection, and relates to a device for preparing a multi-sample single-cell suspension slide microarray.The device comprises a metal slide clamp, the metal slide clamp comprising a base plate connected in rotation and a metal elastic clamping piece, the base plate being fixed with a fixed hook for limiting the metal elastic clamping piece; the base plate and the metal elastic clamping piece clamping and fixing a glass slide, a water-absorbing filter paper and a cell funnel; the cell funnel comprising an inner and outer sample adding area and a mounting area, the thickness of the sample adding area being greater than that of the mounting area, the sample adding area extending outward and protruding away from the water-absorbing filter paper, and the mounting area being used for abutting against the metal elastic clamping piece; the sample adding area being provided with at least six sample adding holes arranged in a matrix; the water-absorbing filter paper being provided with through holes corresponding to the sample adding holes one by one, and the adjacent sample adding holes being separated by the water-absorbing filter paper; and the through holes being opposite to the sample adding holes when the metal slide clamp clamps and fixes the glass slide, the water-absorbing filter paper and the cell funnel. The device can absorb the excess liquid in the centrifugal process through the water-absorbing filter paper. The combination of the water-absorbing filter paper and the cell funnel can simultaneously realize the pre-enrichment treatment of a small amount of cells of multi samples, and can efficiently and accurately prepare the multi-sample single-cell suspension slide microarray, and is suitable for various single-cell sample analysis and research applications.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cytological detection, and in particular to a device for preparing a liquid sample. BACKGROUND

[0002] Existing devices for preparing cytological samples include disposable cell funnels, which are mainly used for large liquid samples to pre-enrich cells onto a slide with an area of up to 15mm x 20mm. For example, see the device disclosed in CN216082239U, a centrifugal liquid-based cell preparation machine. This device can only pre-enrich one liquid sample on one slide at a time, and requires a high amount of cells in the sample, which is not suitable for clinical samples with small volumes or low cell counts. Since there is no filter card between the device and the slide, excess liquid cannot be removed during centrifugation. During the process of removing the slide after centrifugation, the residual liquid in the cell funnel can easily flow back and wash the sample area on the slide, making it impossible to obtain a high-quality single-layer cell area with random distribution, dryness, and clear boundaries. This method is not conducive to the rapid and efficient processing of a large number of cell suspension samples, and batch errors are inevitable if multiple cell suspension samples are detected in batches. With the increasing application of second-generation sequencing technology, this method combined with second-generation sequencing also has the disadvantages of high sequencing cost, long time consumption, high reagent consumption, and large batch effect.

[0003] Tissue Microarray (TMA) is a current slide-based multi-sample cell in situ detection method that can achieve a multi-well microarray arrangement on tissue samples. The manufacturing method of tissue chips can be divided into two types: paraffin-embedded tissue chips and frozen chips. The commonly used type in tumors is paraffin-embedded tissue chips, which are made by a tissue chip manufacturing machine using a fine needle punching method to collect dozens to hundreds of cylindrical small tissues (tissue cores) from numerous tissue wax blocks (called donor wax blocks), and then arrange them in another blank wax block (called the receptor wax block) to form a tissue chip wax block. Then, the tissue chip wax block is sliced, and the slices are transferred to a glass slide to make a tissue chip for in situ detection and analysis.

[0004] Based on the existing TMA technology, which is only applicable to solid tumors or tissue samples, it has not been applied to blood / bone marrow liquid samples and other body fluid samples such as cerebrospinal fluid, urine (urine eosinophil, urine hemoferrin), etc. for clinical microscopic examination and subsequent second-generation sequencing-related omics research. SUMMARY

[0005] To solve the above-mentioned at least one technical problem, the present application provides a device for preparing a multi-sample single-cell suspension slide microarray.

[0006] The technical scheme of the present application is: a device for preparing a multi-sample single cell suspension slide microarray, comprising a metal slide clamp, characterized in that the metal slide clamp comprises a base plate connected in rotation and a metal elastic clamping piece, the base plate is fixed with a fixed hook for limiting the metal elastic clamping piece;

[0007] The base plate and the metal elastic clamping piece clamp and fix the glass slide, the water absorption filter paper and the cell funnel;

[0008] The cell funnel comprises an inner and outer sample adding area and a mounting area, the thickness of the sample adding area is greater than that of the mounting area, the sample adding area extends outwardly and protrudes away from the water absorption filter paper side, and the mounting area is used for abutting against the metal elastic clamping piece; the sample adding area is provided with at least six sample adding holes arranged in a matrix;

[0009] The water absorption filter paper is provided with through holes corresponding to the sample adding holes, and the adjacent sample adding holes are separated by the water absorption filter paper;

[0010] When the metal slide clamp clamps and fixes the glass slide, the water absorption filter paper and the cell funnel, the through holes are opposite to the sample adding holes.

[0011] The water absorption filter paper can absorb excess liquid during centrifugation. The metal slide clamp forms a locking mechanism through the fixed hook of the metal elastic clamping piece, so that the glass slide, the water absorption filter paper and the cell funnel are accurately aligned with the holes during centrifugation, and the single cell suspension can smoothly pass through the sample adding holes and the water absorption filter paper in turn, and finally be adsorbed on the glass slide.

[0012] The combination of the water absorption filter paper and the cell funnel can simultaneously realize the pre-enrichment treatment of a small amount of cells of multiple samples, and can efficiently and accurately prepare a multi-sample single cell suspension slide microarray, which is suitable for various single cell sample analysis and research applications.

[0013] The present application realizes the enrichment of multiple relatively independent sample regions on one glass slide, facilitates the comparative detection of samples on the same glass slide under the same experimental conditions, greatly improves the data interference caused by the experimental condition difference of a single glass slide single sample, and maximally reduces the batch effect between samples. At the same time, the present application greatly improves the sample processing efficiency of the glass slide through the multi-sample enrichment form.

[0014] Further preferably, the cell funnel is a colorless transparent resin material. The volume of the single cell suspension in all sample adding holes before centrifugation can be directly observed by naked eye and recycled multiple times.

[0015] Further preferably, the cell funnel is made of SLA photosensitive resin 3D printing. The resin material has low viscosity, which ensures that the cell suspension is not easily adsorbed on the inner wall, and is convenient for cleaning and recycling. The volume of single cell suspension in all sample wells before centrifugation can be directly observed by naked eye, and the cell funnel can tolerate the pressure of metal slide clamp without deformation.

[0016] Further preferably, the sample well includes a cylindrical hole and a hemispherical hole bottom connected in a butt joint;

[0017] The cylindrical hole of the cell funnel is in one-to-one butt joint with the through hole of the water-absorbing filter paper;

[0018] The inner diameter of the cylindrical hole of the cell funnel matches the inner diameter of the through hole of the water-absorbing filter paper.

[0019] Further preferably, the sample well is provided with six or twelve sample wells.

[0020] Further preferably, when the sample well is six, the inner diameter of the sample well is 4mm, and the inner diameter of the through hole is 4mm;

[0021] When the sample well is twelve, the inner diameter of the sample well is 5mm, and the inner diameter of the through hole is 5mm.

[0022] Further preferably, the outer contour of the mounting area matches the outer contour of the water-absorbing filter paper.

[0023] It is convenient for alignment and coincidence.

[0024] Further preferably, the glass slide is an adhesive type glass slide;

[0025] The glass slide includes a marking area and a sample area, and the two sides of the marking area in the thickness direction are a smooth surface and a frosted surface, respectively, the smooth surface is close to the substrate, and the frosted surface faces the water-absorbing filter paper;

[0026] The sample area is close to the water-absorbing filter paper.

[0027] Further preferably, the size of the water-absorbing filter paper is 25mm×57mm;

[0028] The size of the cell funnel close to the water-absorbing filter paper is 25mm×57mm.

[0029] Further preferably, the mounting area is provided with sample identification, and the sample identification is provided one-to-one with the sample well. The sample identification can be a digital identification or an English identification. The sample identification is a carved sample identification. Through the orientation of the sample identification, the orientation of the cell funnel is facilitated. The mounting area is provided with an orientation mark for installation. The length of the mounting area in the upper region of the sample area is greater than the length of the mounting area in the lower region of the sample area.

[0030] Further preferably, the sample wells are arranged in two rows in a matrix.

[0031] Further preferably, the water-absorbing filter paper is cotton, with a smooth surface and uniform texture.

[0032] Further preferably, the water-absorbing filter paper has a thickness of 2 mm.

[0033] Further preferably, the water-absorbing filter paper is pre-punched by a puncher to form the through holes. The through holes have a diameter and a position corresponding to the sample wells. The through holes have no burrs and clear edges, which do not affect the distribution of cells on the slide when passing through the filter and play a buffering and supporting role.

[0034] Further preferably, the base plate is provided with outwardly protruding positioning shafts on both sides in the width direction.

[0035] A device for preparing a multi-sample single-cell suspension slide microarray, comprising a metal slide clamp, characterized in that the metal slide clamp comprises a rotationally connected base plate and a metal elastic clamping piece, adjacent four sides of the base plate are respectively fixed with slide side limiting plates and slide end limiting plates, and the slide side limiting plates are fixed with fixed hooks for limiting the metal elastic clamping piece.

[0036] The slide side limiting plates and the slide end limiting plates are respectively used for sliding and limiting the slide along the length direction.

[0037] The base plate and the metal elastic clamping piece clamp and fix the slide, the water-absorbing filter paper, and the cell funnel.

[0038] The cell funnel comprises an inner and outer sample adding area and a mounting area, the thickness of the sample adding area is greater than that of the mounting area, the sample adding area extends outwardly and protrudes away from the water-absorbing filter paper, and the mounting area is used for abutting against the metal elastic clamping piece; the sample adding area is provided with at least six matrix-arranged sample wells.

[0039] The water-absorbing filter paper is provided with through holes corresponding to the sample wells one by one, and the water-absorbing filter paper separates adjacent sample wells; the outer contour of the water-absorbing filter paper matches the outer contour of the mounting area.

[0040] The slide is an adherent slide.

[0041] The slide comprises a marking area and a sample area, two sides of the marking area in the thickness direction are respectively a smooth surface and a frosted surface, the smooth surface is used for closely abutting against the base plate, and the frosted surface faces the water-absorbing filter paper.

[0042] The sample area is used to tightly adhere to the water-absorbing filter paper, and the outer contour of the sample area matches the outer contour of the water-absorbing filter paper.

[0043] When the metal slide clamp clamps and fixes the slide, the water-absorbing filter paper and the cell funnel, the through hole is opposite to the sample hole; and the base plate is provided with an open area for being opposite to the sample hole.

[0044] Further preferably, in use, after a certain amount of single-cell suspension is added to the cell funnel, the opening of the sample hole is placed upward, and the liquid is caused to flow into the bottom of the sample hole by gently shaking;

[0045] After the water-absorbing filter paper and the slide are sequentially covered on the cell funnel from bottom to top, the metal slide clamp clamps and fixes the slide, the water-absorbing filter paper and the cell funnel; and the opening of the sample hole is placed upward into a centrifuge for centrifugation.

[0046] During the centrifugation, under the action of centrifugal force, the single-cell suspension passes through the water-absorbing filter paper, the excess PBS solution except for the cells can be buffered and adsorbed by the water-absorbing filter paper, the cells reach the slide and are adsorbed, and a circularly distributed single-cell layer consistent with the shape of the through hole of the water-absorbing filter paper is formed on the slide; as the buffer is consumed, the cells in the spinning area are further air-dried by continuing the centrifugation, and the dry slide microarray with uniform distribution of multiple samples can be obtained by taking out the slide after the centrifugation.

[0047] Advantages:

[0048] 1) In combination with various second-generation sequencing technologies, a small amount of single-cell suspension can be pre-enriched, and multi-dimensional detection of genome, transcriptome, proteome or even subcellular level of multiple samples can be realized on the same slide, the experimental conditions of the samples on the same slide are completely consistent, good quality control between samples can be realized, the repeatability is high, and the sequencing cost, time and reagents are saved.

[0049] 2) The Cytospin Microarray (CMA) obtained by the device can realize detection of various liquid samples such as blood, bone marrow fluid, urine and tissue cell suspension by using a small volume / cell amount of clinical sample, and improve the clinical diagnosis efficiency of the slide in situ detection technology for various diseases, especially blood system tumors.

[0050] 3) The device has the advantages of large amount of single processing sample, simple operation, small amount of sample required and low cost.

[0051] 4) In a specific glass slide area, a small amount of cell suspension pre-enrichment of multiple samples, which can be combined with RNA high-throughput fluorescent hybridization technology (such as Nanostring CosMx), spatial panoramic atlas staining (MACSima) for application, with obvious advantages such as large scale, high throughput, standardization, etc.

[0052] 5) Can be combined with other conventional technologies such as Wright-Giemsa staining, immunohistochemistry (IHC), in situ hybridization (ISH), fluorescence in situ hybridization (FISH), in situ PCR, and various second-generation sequencing technologies such as RNA high-throughput fluorescent hybridization technology (such as Nanostring CosMx), spatial panoramic atlas staining (MACSima) for application, so as to realize the multi-dimensional characterization information of thousands of cells in the genome, transcriptome, proteome or even subcellular level in a single cell suspension specimen, which has the advantages of large scale, high throughput, standardization compared to traditional tissue sections, and at the same time, due to the same experimental conditions of the microarray samples on the same glass slide, excellent experimental quality control can be achieved, and time and reagents can be saved. BRIEF DESCRIPTION OF DRAWINGS

[0053] Figure 1 It is a decomposition diagram for embodiment 1 of the present application;

[0054] Figure 2 It is a structural schematic diagram of the cell funnel of embodiment 1 of the present application;

[0055] Figure 3 It is a perspective view of the cell funnel of embodiment 1 of the present application;

[0056] Figure 4 It is a perspective view of the cell funnel of embodiment 1 of the present application;

[0057] Figure 5 It is a sectional view of the cell funnel of embodiment 1 of the present application;

[0058] Figure 6 It is a microscope schematic diagram of the adjacent sample well corresponding area of the centrifuged glass slide of embodiment 1 of the present application.

[0059] In the figure, 1 is a metal slide clamp, 11 is a base plate, 12 is a fixing hook, 13 is a metal elastic clamping piece, 2 is a glass slide, 3 is an absorbent filter paper, 4 is a cell funnel, 41 is a mounting area, 42 is a sample adding area, 43 is a sample well, 431 is a cylindrical hole, and 432 is a semispherical hole bottom. DETAILED DESCRIPTION

[0060] Reference Figures 1 to 6, specific embodiment 1, a device for preparing a multi-sample single cell suspension slide microarray, comprising a metal slide clamp 1, the metal slide clamp 1 comprising a base plate 11 connected in rotation and a metal elastic clamping piece 13, the base plate 11 being fixed with a fixed hook 12 for limiting the metal elastic clamping piece 13; the base plate 11 and the metal elastic clamping piece 13 clamping and fixing a glass slide 2, a water absorption filter paper 3 and a cell funnel 4; the cell funnel 4 comprising an inner and outer sample adding area 42 and a mounting area 41, the thickness of the sample adding area 42 being greater than the thickness of the mounting area 41, the sample adding area 42 extending outwardly and protruding away from the water absorption filter paper 3, and the mounting area 41 being used for abutting against the metal elastic clamping piece 13; the sample adding area 42 being provided with at least six matrix arranged sample adding holes 43; the water absorption filter paper 3 being provided with through holes corresponding to the sample adding holes 43, and the adjacent sample adding holes 43 being separated by the water absorption filter paper 3; when the metal slide clamp 1 clamps and fixes the glass slide 2, the water absorption filter paper 3 and the cell funnel 4, the through holes are opposite to the sample adding holes 43. The water absorption filter paper 3 can absorb the excess liquid during centrifugation. The metal slide clamp 1 forms a locking mechanism through the fixed hook 12 of the metal elastic clamping piece 13, so as to ensure that the glass slide 2, the water absorption filter paper 3 and the cell funnel 4 are accurately opposite to the holes during centrifugation, and the single cell suspension can smoothly pass through the sample adding holes 43 and the water absorption filter paper 3 in sequence and finally be adsorbed on the glass slide 2. The combination of the water absorption filter paper 3 and the cell funnel 4 can realize the pre-enrichment treatment of a small amount of cells of multiple samples at the same time, and can efficiently and accurately prepare a multi-sample single cell suspension slide microarray, and is suitable for various single cell sample analysis and research applications.

[0061] The cell funnel 4 is a colorless and transparent resin material. The volume of the single cell suspension in all sample adding holes 43 before centrifugation can be directly observed by naked eye and recycled multiple times.

[0062] The cell funnel 4 is made of SLA photosensitive resin 3D printing. The resin material has low viscosity, so that the cell suspension is not easily adsorbed on the inner wall, facilitating cleaning and recycling. The volume of the single cell suspension in all sample adding holes 43 before centrifugation can be directly observed by naked eye, and the cell funnel 4 can resist the pressure of the metal slide clamp 1 without deformation.

[0063] The sample adding hole 43 comprises a cylindrical hole 431 and a hemispherical hole bottom 432 connected in direct communication; the cylindrical hole 431 of the cell funnel 4 is in direct communication with the through hole of the water absorption filter paper 3; the inner diameter of the cylindrical hole 431 of the cell funnel 4 matches the inner diameter of the through hole of the water absorption filter paper 3.

[0064] The sample adding hole 43 is provided with six or twelve sample adding holes.

[0065] When the sample adding hole 43 is six, the inner diameter of the sample adding hole 43 is 4mm, and the inner diameter of the through hole is 4mm; when the sample adding hole 43 is twelve, the inner diameter of the sample adding hole 43 is 5mm, and the inner diameter of the through hole is 5mm.

[0066] The outer contour of the installation area 41 matches the outer contour of the water-absorbing filter paper 3. It is convenient to align and coincide.

[0067] The glass slide 2 is an adherent type glass slide; the glass slide 2 includes a marking area and a sample area, the two sides of the marking area in the thickness direction are a smooth surface and a frosted surface respectively, the smooth surface is close to the base plate 11, and the frosted surface faces the water-absorbing filter paper 3; the sample area is close to the water-absorbing filter paper 3.

[0068] The size of the water-absorbing filter paper 3 is 25mm×57mm; the size of the cell funnel 4 close to the water-absorbing filter paper 3 is 25mm×57mm.

[0069] Referring to Figure 5 , the installation area 41 is provided with sample marks, and the sample marks are arranged one by one with the sample wells 43. The sample marks can be numerical marks or English marks. The sample marks are engraved sample marks. Through the orientation of the sample marks, the orientation of the cell funnel is convenient to position. The length of the installation area in the upper area of the sample area is greater than the length of the installation area in the lower area of the sample area. Here, up and down are defined based on Figure 1 the perspective.

[0070] The sample wells 43 are arranged in a two-row matrix. Alternatively, they are arranged in a three-row matrix.

[0071] The water-absorbing filter paper 3 is cotton. The surface is flat and the texture is uniform. The thickness of the water-absorbing filter paper 3 is 2mm. The water-absorbing filter paper 3 is pre-punched to form a through hole by a punch. The hole diameter and position of the through hole are arranged corresponding to the sample wells 43. There is no burr in the through hole, and the edge is clear, which does not affect the distribution of cells passing through to the glass slide 2, and plays a buffering and supporting filtering role.

[0072] The base plate is provided with a hollow area opposite to the sample wells. It is convenient to visually observe the cell adhesion on the glass slide. Further determine whether the centrifugation process needs to continue. Once the outline of the multiple circular areas on the glass slide can be seen through the hollow area, the centrifugation process can be stopped. Otherwise, continue centrifugation.

[0073] The metal elastic clamping piece includes a bent metal wire, the metal wire includes a crimping part arranged side by side, the crimping part abuts against the installation area, the metal wire includes an expanding part with increasing distance from the crimping part, and the expanding part is used for clamping into the fixing hook. The expanding part is bent away from the base plate relative to the crimping part.

[0074] The base plate 11 is provided with an outwardly protruding positioning shaft on both sides in the width direction. The positioning shaft is convenient to cooperate with the turntable of the centrifuge.

[0075] In use, after a certain amount of single-cell suspension is added to the cell funnel 4, the opening of the sample well 43 is placed upward, and the liquid is shaken to flow into the bottom of the sample well 43;

[0076] After sequentially covering the water-absorbing filter paper 3 and the glass slide 2 on the cell funnel 4 from bottom to top, the metal glass slide clamp 1 clamps and fixes the glass slide 2, the water-absorbing filter paper 3 and the cell funnel 4; the opening of the sample adding hole 43 is upwardly placed into the centrifuge for centrifugation. At this time, the glass slide is obliquely arranged.

[0077] During the centrifugation, under the action of centrifugal force, the single cell suspension passes through the water-absorbing filter paper 3, and the excess PBS solution outside the cells can be buffered and adsorbed by the water-absorbing filter paper 3, and the cells reach the glass slide 2 and are adsorbed, forming a circularly distributed single cell layer on the glass slide 2 consistent with the shape of the through hole of the water-absorbing filter paper 3; as the buffer is consumed, continuing to centrifuge will further dry the cells in the spinning area, and the dry multi-sample uniformly arranged glass slide microarray can be obtained by taking out the glass slide after centrifugation. See Figure 6 , there is no mutual interference between the adjacent samples on the centrifuged glass slide. The separation is obvious.

[0078] Specific embodiment 2, a device for preparing a multi-sample single cell suspension glass slide microarray, comprising a metal glass slide clamp 1, characterized in that the metal glass slide clamp 1 comprises a base plate and a metal elastic clamping piece connected by rotation, the adjacent four sides of the base plate are respectively fixed with glass slide side limiting plates and glass slide end limiting plates, the glass slide side limiting plates are fixed with fixed hooks for limiting the metal elastic clamping piece; the glass slide side limiting plates and the glass slide end limiting plates are respectively used for sliding and limiting the glass slide along the length direction; the base plate and the metal elastic clamping piece clamp and fix the glass slide 2, the water-absorbing filter paper 3 and the cell funnel 4; the cell funnel 4 comprises an inner and outer sample adding area and a mounting area, the thickness of the sample adding area is greater than that of the mounting area, the sample adding area extends outwardly and protrudes away from the water-absorbing filter paper 3, and the mounting area is used for abutting against the metal elastic clamping piece; the sample adding area is provided with at least six sample adding holes arranged in a matrix; the water-absorbing filter paper 3 is provided with through holes corresponding to the sample adding holes one by one, and the adjacent sample adding holes are separated by the water-absorbing filter paper 3; the outer contour of the water-absorbing filter paper 3 matches the outer contour of the mounting area; the glass slide 2 is an adhesive type glass slide; the glass slide 2 comprises a marking area and a sample area, the two sides of the marking area in the thickness direction are respectively a smooth surface and a frosted surface, the smooth surface is used for closely adhering to the base plate, and the frosted surface faces the water-absorbing filter paper 3; the sample area is used for closely adhering to the water-absorbing filter paper 3, and the outer contour of the sample area matches the outer contour of the water-absorbing filter paper 3; when the metal glass slide clamp 1 clamps and fixes the glass slide 2, the water-absorbing filter paper 3 and the cell funnel 4, the through holes are opposite to the sample adding holes; the base plate is provided with a hollow area, and the hollow area is used for being opposite to the sample adding holes. The hollow area is opposite to all the sample adding holes. Alternatively, the hollow area is opposite to the sample adding holes one by one.

[0079] See Figure 4 When the sample adding hole 43 is six, the inner diameter of the sample adding hole 43 is 4mm, and the inner diameter of the through hole is 4mm.

[0080] The size of the absorbent filter paper 33 is 25 mm×57 mm; the size of the cell funnel 44 close to the absorbent filter paper 33 is 25 mm×57 mm.

[0081] The length (up and down) of the sample loading area is 20mm, and the width (front and back) is 15mm. The length from the bottom of the sample loading area to the bottom of the installation area is 17mm. The distance from the front and back sides of the sample loading area to the front and back sides of the installation area is 5mm. The up and down positions here are based on Figure 1 The mark area and the sample area are defined by the viewing angle.

[0082] The depth of the sample well is 30 mm, and the thickness of the sample area along the axial direction of the sample well is 31 mm.

[0083] Specific embodiment 3, a device for preparing a multi-sample single-cell suspension glass slide microarray, comprising a metal glass slide clamp 1, characterized in that the metal glass slide clamp 1 comprises a rotatably connected base plate and a metal elastic clamping member, the four adjacent sides of the base plate are respectively fixed with a slide side limiting plate and a slide end limiting plate, the slide side limiting plate is fixed with a fixing hook for limiting the metal elastic clamping member; the slide side limiting plate and the slide end limiting plate are respectively used for the slide to slide in and limit along the length direction; the base plate and the metal elastic clamping member clamp and fix the slide 2, the absorbent filter paper 3 and the cell funnel 4; the cell funnel 4 comprises a sample adding area and an installation area arranged inside and outside, the thickness of the sample adding area is greater than the thickness of the installation area, the sample adding area extends outwardly away from the absorbent filter paper 3, and the installation area is used The sample loading area is provided with at least six sample loading holes arranged in a matrix; the absorbent filter paper 3 is provided with through holes arranged in a one-to-one correspondence with the sample loading holes, and the adjacent sample loading holes are separated by the absorbent filter paper 3; the outer contour of the absorbent filter paper 3 matches the outer contour of the mounting area; the glass slide 2 is an adhesive glass slide; the glass slide 2 includes a marking area and a sample area, and the two sides of the marking area in the thickness direction are respectively a smooth surface and a frosted surface, the smooth surface is used to be close to the substrate, and the frosted surface faces the absorbent filter paper 3; the sample area is used to be close to the absorbent filter paper 3, and the outer contour of the sample area matches the outer contour of the absorbent filter paper 3; when the metal slide clamp 1 clamps and fixes the glass slide 2, the absorbent filter paper 3 and the cell funnel 4, the through holes are opposite to the sample loading holes; a hollow area is provided on the substrate, and the hollow area is used to face the sample loading holes.

[0084] See also Figure 3 When there are twelve sample addition holes 43, the inner diameter of the sample addition holes 43 is 5 mm, and the inner diameter of the through hole is 5 mm.

[0085] The size of the absorbent filter paper 3 is 25 mm×57 mm; the size of the cell funnel 4 close to the absorbent filter paper 3 is 25 mm×57 mm.

[0086] The length (in the up-down direction) of the sample application area is 20 mm, and the width (in the front-back direction) is 15 mm. The length from the lower side of the sample application area to the bottom of the mounting area is 17 mm. The distance from the front and back sides of the sample application area to the front and back sides of the mounting area is 5 mm. The up-down direction here is defined based on the angle of view. The mark area and the sample area are arranged up and down. Figure 1 The mark area and the sample area are arranged up and down.

[0087] The depth of the sample application hole is 30 mm. The thickness of the sample application area along the axial direction of the sample application hole is 31 mm.

[0088] The above is only a preferred embodiment of the present application, and it should be noted that those of ordinary skill in the art can make several improvements and refinements without departing from the principles of the present application, and these improvements and refinements should also be considered within the scope of protection of the present application.

Claims

1. A device for preparing a multi-sample single cell suspension slide microarray, comprising a metal slide holder, characterized in that: The metal slide clamp comprises a base plate and a metal elastic clamping member which are rotatably connected, and a fixing hook for limiting the metal elastic clamping member is fixed on the base plate; The glass slide, absorbent filter paper and cell funnel are clamped and fixed between the substrate and the metal elastic clamping member; The cell funnel includes a sample loading area and a mounting area provided inside and outside. The sample loading area is thicker than the mounting area. The sample loading area extends outward away from the absorbent filter paper, and the mounting area is used to abut against the metal elastic clamping member. The sample loading area is provided with at least six sample loading wells arranged in a matrix. The sample loading wells are blind holes with their openings facing the slide. The absorbent filter paper is provided with through holes corresponding to the sample addition wells one by one, and adjacent sample addition wells are separated by the absorbent filter paper; When the metal slide clamp clamps and fixes the glass slide, the absorbent filter paper and the cell funnel, the through hole faces the sample addition hole; The length of the installation area above the sample area is greater than the length of the installation area below the sample area; The absorbent filter paper is made of cotton; The thickness of the absorbent filter paper is 2 mm; The absorbent filter paper is pre-punched by a puncher to form the through holes; The length of the sample loading area is 20mm and the width is 15mm; the length from the bottom of the sample loading area to the bottom of the installation area is 17mm; the distance from the front and back sides of the sample loading area to the front and back sides of the installation area is 5mm; When in use, after adding a certain amount of single cell suspension to the cell funnel, place the opening of the sample well upwards and gently shake it to allow the liquid to flow into the bottom of the sample well. After covering the cell funnel with absorbent filter paper and a glass slide from bottom to top, a metal slide clamp fixes the glass slide, the absorbent filter paper and the cell funnel; the cell funnel is placed in a centrifuge with the opening of the sample well facing upward for centrifugation, and the glass slide is set at an angle.

2. The device for preparing a multi-sample single cell suspension slide microarray according to claim 1, characterized in that: The cell funnel is a colorless and transparent resin material; The cell funnel is made by SLA photosensitive resin 3D printing.

3. The device for preparing a multi-sample single cell suspension slide microarray according to claim 1, characterized in that: The sample addition hole comprises a cylindrical hole connected to each other and a hemispherical hole bottom; The cylindrical holes of the cell funnel are connected one by one with the through holes of the absorbent filter paper; The inner diameter of the cylindrical hole of the cell funnel matches the inner diameter of the through hole of the absorbent filter paper.

4. The device for preparing a multi-sample single cell suspension slide microarray according to claim 1, characterized in that: The number of the sample addition wells is six or twelve.

5. The device for preparing a multi-sample single cell suspension slide microarray according to claim 1, characterized in that: When there are six sample addition holes, the inner diameter of the sample addition holes is 4 mm, and the inner diameter of the through hole is 4 mm; When there are twelve sample addition holes, the inner diameter of the sample addition holes is 5 mm, and the inner diameter of the through hole is 5 mm.

6. The device for preparing a multi-sample single cell suspension slide microarray according to claim 1, characterized in that: The outer contour of the installation area matches the outer contour of the water-absorbing filter paper; The installation area is provided with sample identifiers, and the sample identifiers are provided in a one-to-one correspondence with the sample addition holes.

7. The device for preparing a multi-sample single cell suspension slide microarray according to claim 1, characterized in that: The slide glass is an adhesive slide; The glass slide comprises a marking area and a sample area, wherein two sides of the marking area in the thickness direction are respectively a smooth surface and a frosted surface, the smooth surface is in close contact with the substrate, and the frosted surface faces the absorbent filter paper; The sample area is in close contact with the absorbent filter paper.

8. A device for preparing a multi-sample single cell suspension slide microarray, comprising a metal slide holder, characterized in that: The metal slide clamp comprises a rotatably connected base plate and a metal elastic clamping member, wherein four adjacent sides of the base plate are respectively fixed with a slide side limiting plate and a slide end limiting plate, and the slide side limiting plate is fixed with a fixing hook for limiting the metal elastic clamping member; The side limiting plates of the glass slide and the end limiting plates of the glass slide are used for sliding the glass slide in and limiting the position along the length direction respectively; The glass slide, absorbent filter paper and cell funnel are clamped and fixed between the substrate and the metal elastic clamping member; The cell funnel includes a sample loading area and a mounting area provided inside and outside. The sample loading area is thicker than the mounting area. The sample loading area extends outward away from the absorbent filter paper, and the mounting area is used to abut against the metal elastic clamping member. The sample loading area is provided with at least six sample loading wells arranged in a matrix. The sample loading wells are blind holes with their openings facing the slide. The absorbent filter paper is provided with through holes corresponding to the sample addition holes one by one, and adjacent sample addition holes are separated by the absorbent filter paper; the outer contour of the absorbent filter paper matches the outer contour of the installation area; The slide glass is an adhesive slide; The glass slide comprises a marking area and a sample area, wherein two sides of the marking area in the thickness direction are respectively a smooth surface and a frosted surface, wherein the smooth surface is used to be in close contact with the substrate, and the frosted surface faces the absorbent filter paper; The sample area is used to be in close contact with the water-absorbing filter paper, and the outer contour of the sample area matches the outer contour of the water-absorbing filter paper; When the metal slide clamp clamps the glass slide, the absorbent filter paper and the cell funnel, the through hole faces the sample loading hole; a hollow area is provided on the substrate, and the hollow area is used to face the sample loading hole; The length of the installation area above the sample area is greater than the length of the installation area below the sample area; The absorbent filter paper is made of cotton; The thickness of the absorbent filter paper is 2 mm; The absorbent filter paper is pre-punched by a puncher to form the through holes; The length of the sample loading area is 20mm and the width is 15mm; the length from the bottom of the sample loading area to the bottom of the installation area is 17mm; the distance from the front and back sides of the sample loading area to the front and back sides of the installation area is 5mm; When in use, after adding a certain amount of single cell suspension to the cell funnel, place the opening of the sample well upwards and gently shake it to allow the liquid to flow into the bottom of the sample well. After covering the cell funnel with absorbent filter paper and a glass slide from bottom to top, a metal slide clamp fixes the glass slide, the absorbent filter paper and the cell funnel; the cell funnel is placed in a centrifuge with the opening of the sample well facing upward for centrifugation; and the glass slide is tilted.

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