A flexible slide-based digital microscopy analysis method

Abstract

The application relates to a flexible-slit-based digital microscopic analysis method; the flexible-slit-based digital microscopic analysis method is provided, and the steps include the following: S1, providing a flexible slit, the flexible slit comprises a slit main body, a connecting part and a supporting part; the slit main body, the connecting part and the supporting part surround a cavity; S2, adding a to-be-detected object containing a biological sample to the cavity to realize enrichment of the biological sample; S3, separating the slit main body from the connecting part, and pasting a side of the slit main body, which is specifically adsorbed with the biological sample, to a photosensitive surface of an image sensing chip, so that the digital microscopic analysis is completed; in the application, the slit main body of the flexible slit can be pasted to a rigid material plane; high light transmittance and smoothness make the microscopic image of the observation target have no distortion phenomenon; the flexible slit as a whole has chemical inertia and does not participate in related reactions related to surface enrichment substances; the slit main body can be separated from the connecting part, and the operation is simple.

Description

Technical Field

[0001] This invention relates to the field of bioanalysis, and more particularly to a digital microscopy method based on flexible glass slides. Background Technology

[0002] Recording full-field microscopic images of the material under study on a glass slide using image sensor chips, and then transmitting these images to a computer for storage, display, analysis, and diagnosis, constitutes the technical framework for digital microscopy clinical diagnosis and is an important component of digital healthcare. Image sensor chip-based microscopy is a lens-free digital microscopy technique. High-resolution, wide-field-of-view image sensor chips can directly record full-field microscopic images of the material under study on a glass slide. However, this requires the distance between the recorded material and the surface of the image sensor chip to be within tens of micrometers. Existing glass slides, due to their rigidity, cannot meet this distance requirement.

[0003] A glass slide is a transparent, planar device that carries the substances being observed. In the life sciences and clinical medicine, slides are used to hold cells, pathogens, and tissue sections. Various visible light / fluorescence staining and antigen-antibody reactions are then performed on the slides, followed by microscopic observation under a microscope for life science research and clinical diagnosis. Slides are generally made of glass or quartz, possessing high light transmittance and a smooth surface to ensure clear microscopic observation. They are typically rectangular pieces with a thickness in the millimeter range, exhibiting high rigidity, non-deformation, and ease of handling. The center of the slide is the sample area, which holds the observed sample; its area constitutes the entire field of view required for microscope observation. The process of securely placing the enriched and purified substances from the sample into the sample area of ​​the slide is called slide preparation. After preparation, there is a need for staining and on-slide biochemical reactions. The entire slide preparation, staining, and on-slide biochemical process is complex, involving a significant amount of manual operation and low levels of automation, resulting in inconsistent slide quality. Summary of the Invention

[0004] The purpose of this invention is to address the shortcomings of existing technologies by providing a digital microscopy analysis method based on flexible glass slides.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] A digital microscopy analysis method based on flexible glass slides is provided, comprising the following steps:

[0007] S1. A flexible glass slide is provided, the flexible glass slide comprising:

[0008] The glass slide body is used for the specific adsorption of biological samples;

[0009] A connecting portion is fixedly disposed on the outer periphery of the glass slide body;

[0010] A support portion, the first end of which is fixedly disposed on the outer periphery of the connecting portion, and the second end of which is an open end, the slide body, the connecting portion, and the support portion forming a cavity;

[0011] S2. The analyte containing the biological sample is added to the chamber, so that the slide body specifically adsorbs the biological sample, thereby enriching the biological sample;

[0012] S3. After removing the analyte that was not specifically adsorbed by the slide body, separate the slide body from the connecting part, and attach the side of the slide body that specifically adsorbs the biological sample to the photosensitive surface of the image sensor chip to complete the digital microscopic analysis.

[0013] Preferably, the slide body has several modified portions on one side used for specifically adsorbing the biological sample.

[0014] More preferably, the plurality of said modifications include one or more of a plurality of chemical modifications or a plurality of physical modifications.

[0015] More preferably, the plurality of chemical modification portions include one or more of the following: a plurality of electrostatic substance modification portions, a plurality of specific antigen antibody modification portions, or a plurality of gene probe modification portions.

[0016] More preferably, the plurality of said physical modification portions include one or more of a plurality of pit array modification portions or a plurality of liquid flow groove modification portions.

[0017] Preferably, the connecting portion is provided with a plurality of filter structures for removing the analytes that are not specifically adsorbed by the glass slide body.

[0018] Preferably, the flexible glass slide is made of an inert chemical material.

[0019] Preferably, between steps S2 and S3, the method further includes one or more of the following steps: purification of the biological sample, staining of the biological sample, biological testing of the biological sample, or biochemical labeling of the biological sample.

[0020] The present invention adopts the above technical solution and has the following technical effects compared with the prior art:

[0021] The flexible glass slide of this invention has a suitable ratio of flexibility to rigidity, allowing it to conform to a rigid material plane; its high light transmittance and smoothness ensure distortion-free microscopic images of the observed target; the flexible glass slide is chemically inert and does not participate in the reactions involved in the accumulation of substances on the surface; it can be used alone or in conjunction with other equipment as a medical instrument, depending on the needs of different life science research, clinical testing, and clinical pathology diagnosis; the slide body can be detached from the connecting part, making operation simple. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of the basic structure of the flexible glass slide in Embodiment 1 of the present invention;

[0023] Figure 2 This is a schematic diagram of the bottom surface of the flexible glass sheet in Embodiment 1 of the present invention;

[0024] Figure 3 This is a schematic diagram of the bonding between the flexible glass sheet and the image sensing chip in Embodiment 1 of the present invention;

[0025] Figure 4 This is a full-view micrograph of the slide body in Embodiment 1 of the present invention;

[0026] Figure 5 for Figure 4 A magnified view of a portion of the image;

[0027] Figure 6 This is a schematic diagram of the basic structure of the flexible glass slide in Embodiment 2 of the present invention;

[0028] Figure 7 This is a schematic diagram of the bottom surface of the flexible glass sheet in Embodiment 2 of the present invention;

[0029] Figure 8 This is a schematic diagram of the bonding between the flexible glass sheet and the image sensing chip in Embodiment 2 of the present invention;

[0030] Figure 9 This is a full-view micrograph of the slide body in Embodiment 2 of the present invention;

[0031] Figure 10 for Figure 9 A magnified view of a portion of the image;

[0032] The reference numerals in the figure include:

[0033] 1. Glass slide body; 2. Connecting part; 3. Supporting part. Detailed Implementation

[0034] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0035] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0036] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but this is not intended to limit the scope of the invention.

[0037] Example 1

[0038] like Figure 1-5 As shown, this embodiment provides a digital microscopy analysis method based on flexible glass slides, the steps of which include:

[0039] S1. A flexible glass slide is provided, the flexible glass slide comprising:

[0040] The slide body 1 is used to specifically adsorb cervical exfoliated cells; the side of the slide body 1 used to specifically adsorb the cervical exfoliated cells is modified with L-poly-L-lysine.

[0041] A connecting part 2 is fixedly disposed on the outer periphery of the glass slide body 1; the connecting part 2 is provided with a plurality of filter structures;

[0042] The support part 3 has a first end fixedly disposed on the outer periphery of the connecting part 2, and a second end of the support part 3 is an open end. The glass slide body 1, the connecting part 2 and the support part 3 surround a cavity.

[0043] S2. Add the cervical swab washing and preservation solution containing the cervical exfoliated cells to the chamber, so that the slide body 1 specifically adsorbs the cervical exfoliated cells, thereby enriching the cervical exfoliated cells.

[0044] S3. After removing the liquid that was not specifically adsorbed by the slide body 1, separate the slide body 1 from the connecting part 2, and attach the side of the slide body 1 that specifically adsorbed the cervical exfoliated cells to the photosensitive surface of the image sensor chip to complete the digital microscopic analysis.

[0045] Optionally, between steps S2 and S3, the method further includes: staining the cervical exfoliated cells; specifically, the staining solution is dripped into the cavity.

[0046] In a preferred embodiment, the glass slide body 1 is made of flexible glass, and the glass slide body 1 is in the shape of a disc, with a thickness of 100 μm, a diameter of 14 mm, and a light transmittance of more than 90%.

[0047] In a preferred embodiment, the connecting part 2 is made of transparent polycarbonate, and the connecting part 2 is annular, with an outer ring diameter of 18 mm, a ring width of 2 mm, and a thickness of 300 μm;

[0048] In a preferred embodiment, the support portion 3 is made of transparent polypropylene, and the support portion 3 has a height of 60 mm and a diameter of 22 mm;

[0049] In a preferred embodiment, the molecular weight of the poly-L-lysine used for the L-type poly-L-lysine modification is 150,000-300,000. The specific steps include: preparing a 0.1% poly-L-lysine aqueous solution and storing it at -20°C; slowly adding an appropriate amount of poly-L-lysine aqueous solution to the flexible glass slide area until it completely covers the flexible glass slide; and then air-drying it.

[0050] Example 2

[0051] like Figure 6-10 As shown, this embodiment provides another digital microscopy analysis method based on flexible glass slides, the steps of which include:

[0052] S1. A flexible glass slide is provided, the flexible glass slide comprising:

[0053] The slide body 1 is used to specifically adsorb and capture positive magnetic nanospheres of human blood cardiac troponin I (cTn I); one side of the slide body 1 used to specifically adsorb the positive magnetic nanospheres is coated with cTn I secondary antibody.

[0054] Connecting part 2, the connecting part 2 is fixedly disposed on the outer periphery of the glass slide body 1;

[0055] The support part 3 has a first end fixedly disposed on the outer periphery of the connecting part 2, and a second end of the support part 3 is an open end. The glass slide body 1, the connecting part 2 and the support part 3 surround a cavity.

[0056] S2. Add the mixture containing the positive magnetic nanospheres and the negative magnetic nanospheres that have not captured cTnI to the chamber and incubate at 37°C for 15 min to allow the glass slide body 1 to specifically adsorb the positive magnetic nanospheres.

[0057] S3. After removing the analyte that was not specifically adsorbed by the slide body 1, separate the slide body 1 from the connecting part 2, and attach the side of the slide body 1 with the positive magnetic nanospheres specifically adsorbed to the photosensitive surface of the image sensor chip to complete the digital microscopic analysis.

[0058] In a preferred embodiment, the glass slide body 1 is made of colorless and transparent polystyrene material. The glass slide body 1 is in the shape of a disc, with a thickness of 1.5 mm, a diameter of 14 mm, and a light transmittance of more than 90%.

[0059] In a preferred embodiment, the connecting part 2 is made of transparent polycarbonate, and the connecting part 2 is annular, with an outer ring diameter of 18 mm, a ring width of 2 mm, and a thickness of 300 μm;

[0060] In a preferred embodiment, the support portion 3 is made of transparent polypropylene, and the support portion 3 has a height of 10 mm and a diameter of 22 mm;

[0061] In a preferred embodiment, step S1, the step of coating the slide body 1 with cTn I secondary antibody, includes: preparing an antibody solution of 0.526 mg / mL in a 2-(N-morpholino)ethanesulfonic acid (MES) solution with a pH of 6.2; adding sufficient antibody solution to the chamber of the flexible slide so that the antibody solution completely covers the slide body 1, and incubating it in a light-proof environment at 2℃-8℃ for 30 min; placing a solution containing 1 mg / mL EDC in a cold MES solution with a pH of 6.0, and adding this solution to the chamber of the flexible slide, and continuing to incubate in the same environment for 30 min; after incubation, pouring out the solution and washing twice with a mixture of PBS and 0.1% Tween buffer; adding 1% bovine serum albumin PBS solution and incubating it in a light-proof environment at 2℃-8℃ for 60 min; then washing twice with a mixture of PBS and 0.1% Tween buffer, and adding buffer for storage.

[0062] In summary, the flexible glass slide of this invention possesses a suitable ratio of flexibility to rigidity, allowing it to conform to a rigid material plane; its high light transmittance and smoothness ensure distortion-free microscopic images of the observed target; the flexible glass slide as a whole is chemically inert and does not participate in the reactions involved in the accumulation of substances on the surface; it can be used alone or in conjunction with other equipment as a medical instrument, depending on the needs of different life science research, clinical testing, and clinical pathology diagnosis; the slide body can be detached from the connecting part, simplifying operation.

[0063] The above description is merely a preferred embodiment of the present invention and does not limit the implementation and protection scope of the present invention. Those skilled in the art should realize that any equivalent substitutions and obvious changes made based on the description and illustrations of the present invention should be included within the protection scope of the present invention.

Claims

1. A method of digital microscopic analysis based on a flexible glass slide, characterized by the steps of include: S1. A flexible glass slide is provided, the flexible glass slide comprising: The glass slide body (1) is used for the specific adsorption of biological samples; A connecting part (2) is fixedly disposed on the outer periphery of the glass slide body (1); The support part (3) has a first end fixedly disposed on the outer periphery of the connecting part (2) and a second end of the support part (3) being an open end. The glass slide body (1), the connecting part (2) and the support part (3) surround a cavity. S2. The analyte containing the biological sample is added to the chamber, so that the slide body (1) specifically adsorbs the biological sample, thereby enriching the biological sample; S3. After removing the analyte that was not specifically adsorbed by the slide body (1), separate the slide body (1) from the connecting part (2), and attach the side of the slide body (1) that specifically adsorbs the biological sample to the photosensitive surface of the image sensor chip to complete the digital microscopic analysis. The connecting part (2) is provided with a plurality of filter structures for removing the analytes that are not specifically adsorbed by the glass slide body (1).

2. The digital microscopic analysis method of claim 1, wherein, The slide body (1) has several modified parts on one side used for specifically adsorbing the biological sample.

3. The digital microscopic analysis method of claim 2, wherein, The aforementioned modifications include one or more of the following: chemical modifications or physical modifications.

4. The digital microscopic analysis method of claim 3, wherein, The chemical modification portions include one or more of the following: electrostatic substance modification portions, specific antigen-antibody modification portions, or gene probe modification portions.

5. The digital microscopic analysis method of claim 3, wherein, The physical modification portions include one or more of the following: a plurality of pit array modification portions or a plurality of liquid flow groove modification portions.

6. The digital microscopic analysis method of claim 1, wherein, The flexible glass slide is made of inert chemical materials.

7. The digital microscopy analysis method according to claim 1, characterized in that, Between steps S2 and S3, the method further includes one or more of the following steps: purification of the biological sample, staining of the biological sample, biological testing of the biological sample, or biochemical labeling of the biological sample.

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