Microfluidic pathological glass slide chip and application thereof

Abstract

The invention provides a microfluidic pathological glass slide chip and application thereof.The microfluidic pathological glass slide chip comprises a substrate and a cover plate pressed on the substrate, a microchannel is defined by the substrate and the cover plate, a detection area is arranged in the middle of the microchannel, check blocks are arranged in the detection area, the check blocks are distributed on the upper surface of the substrate and the lower surface of the cover plate in a staggered mode, and the check blocks are arranged on the upper surface of the substrate and the lower surface of the cover plate in a staggered mode. One end of the micro-channel is connected with the sample adding hole, the other end of the micro-channel is connected with the driving port, and a water-absorbing material adsorbs liquid in the micro-channel through the driving port. According to the microchannel detection device, visible particles such as crystals or cells in a sample can be enriched to a detection area of the microchannel, the sample does not need to be centrifuged, operation steps are reduced, and inaccurate results caused by reduction of the sample size are also avoided.

Description

Technical Field

[0001] The invention belongs to the technical field of in vitro diagnosis and immune detection, and particularly relates to a microfluidic pathology glass slide chip. Background Art

[0002] Urine sediment testing is a method used to analyze cells, bacteria, crystals, and other tiny particles in urine and is important in the diagnosis and management of urinary tract diseases.

[0003] One of the existing detection methods is to detect through glass slides. During the detection process, the sample needs to be centrifuged, smeared, dried, fixed, and stained. Finally, the glass slide is made and tested under a microscope. This traditional technical method requires the use of a variety of different experimental equipment and multiple steps. The operation is complicated and time-consuming. The slide preparation results have large errors, the samples are easily contaminated, and the requirements for operators are high, which greatly limits its application in grassroots units.

[0004] Microfluidic chips are the main platform for the realization of microfluidic technology. They can integrate basic operation units such as sample preparation, reaction, separation, and detection in biological, chemical, and medical analysis processes into a very small closed chip. Microfluidic chips have the advantages of being lightweight, using less sample and reagent, fast reaction speed, large-scale parallel processing, and disposable. They have great potential in the fields of biology, chemistry, and medicine. In recent years, they have developed into a new research field that intersects biology, chemistry, medicine, fluids, electronics, materials, and mechanics. Summary of the invention

[0005] In view of the shortcomings of existing urine slide detection, the present invention proposes a microfluidic pathology slide chip, which can simplify the urine detection process. Different particulate matter such as cells, bacteria, crystals and other tangible substances can be intercepted, enriched and stained on the same slide chip, which is simple to operate, reduces pollution and saves detection time.

[0006] To achieve the above-mentioned purpose, the technical solution of the present invention is implemented as follows: a microfluidic pathology slide chip includes a substrate and a cover sheet pressed onto the substrate, the substrate and the cover sheet enclose a microchannel, a detection area is provided in the middle of the microchannel, a block is provided in the detection area, the blocks are alternately distributed on the upper surface of the substrate and the lower surface of the cover sheet, one end of the microchannel is connected to the sample loading hole, and the other end is connected to the driving port, and the water-absorbing material absorbs the liquid in the microchannel through the driving port.

[0007] In some embodiments of the present invention, the lower surface of the cover sheet is provided with grooves along its length direction, and the cover sheet and the upper surface of the substrate are enclosed by the grooves to form a microchannel.

[0008] In some embodiments of the present invention, there are multiple blocks, and the interval between two adjacent blocks is 1um-20um. The block is one or both of "cylindrical" and "plate-shaped".

[0009] In some embodiments of the present invention, the height of the block is smaller than the height of the microchannel, and the height of the microchannel is 20um-30um.

[0010] In some embodiments of the present invention, the sample loading hole is disposed on the cover sheet, and the sample loading hole is communicated with the microchannel.

[0011] On the other hand, the present invention also provides an application of using the glass slide chip in any of the above technical solutions for detection, comprising at least the following steps:

[0012] 1) Adding samples: Add the sample to be tested to the sample well, allowing the sample to flow into the detection area, and then contact the water-absorbing material with the driving port to absorb the liquid in the microchannel;

[0013] 2) Elution: Add cleaning solution drops into the sample loading hole to allow the cleaning solution to flow into the detection area, and then bring the absorbent material into contact with the drive port to absorb the liquid in the microchannel.

[0014] On the other hand, the present invention also provides an application of using the glass slide chip in any of the above technical solutions for detection, comprising at least the following steps:

[0015] 1) Adding samples: Add the sample to be tested to the sample well, allowing the sample to flow into the detection area, and then contact the water-absorbing material with the driving port to absorb the liquid in the microchannel;

[0016] 2) Elution: Add the cleaning solution dropwise into the sample loading well to allow the cleaning solution to flow into the detection area, and then contact the absorbent material with the drive port to absorb the liquid in the microchannel;

[0017] 3) Staining: Add the dye solution to the sample well, let the dye solution flow into the detection area, and then contact the water-absorbent material with the driving port to absorb the liquid in the microchannel;

[0018] 4) Elution again: Add the cleaning solution dropwise into the sample loading hole, allow the cleaning solution to flow into the detection area, and then contact the water-absorbent material with the driving port to absorb the liquid in the microchannel;

[0019] 5) Drying: Dry quickly with 37°C dry air or bake in a 37°C oven for 30-60 minutes. The microfluidic pathology glass slide chip obtained by the above technical solution has the following beneficial effects:

[0020] 1. The tangible particles in the sample, such as crystals or cells, are enriched in the detection area of ​​the microchannel, without the need for centrifugation and smearing of the sample, which reduces the number of operation steps and avoids inaccurate results due to reduced sample volume.

[0021] 2. The cleaning, staining and fixation of samples are all integrated into the glass slide. Only the steps of dripping and aspiration need to be repeated, which is simple and quick to operate. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a schematic diagram of the structure of the microfluidic pathology glass slide chip of the present invention;

[0023] Figure 2 1 is a schematic diagram of the side view structure of the microfluidic pathology glass slide chip of the present invention;

[0024] Figure 3 is a schematic structural diagram of the substrate of the present invention;

[0025] Figure 4 is a schematic structural diagram of the cover sheet of the present invention;

[0026] Figure 5 Schematic diagram of the cross-sectional structure of the microfluidic pathology glass slide chip of the present invention;

[0027] Figure 6 It is a schematic structural diagram of another shape of the microfluidic pathology glass slide chip of the present invention.

[0028] In the figure, 1, substrate; 2, cover plate; 3, microchannel; 4, sample loading hole; 5, driving port; 6, water-absorbing material; 31, detection area; 31a, stopper. DETAILED DESCRIPTION

[0029] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0030] Unless otherwise defined, the technical terms used in the following examples have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods, unless otherwise specified, are all conventional methods.

[0031] The present invention generally relates to the field of immunoassay technology, and relates to a microfluidic pathology glass slide chip. The core of the present technical invention is based on microfluidic technology, and a microfluidic pathology glass slide chip is designed to make urine detection more convenient and quick.

[0032] The present invention is further explained below in conjunction with embodiments and drawings. It should be understood that the present invention is not limited to the specific embodiments described.

[0033] like Figure 1 , Figure 2 As shown, the present invention proposes a microfluidic pathology slide chip, including a substrate 1 and a cover sheet 2 pressed onto the substrate 1, the substrate 1 and the cover sheet 2 enclose a microchannel 3, a detection area 31 is arranged in the middle of the microchannel 3, a block 31a is arranged in the detection area 31, the block 31a is staggeredly distributed on the upper surface of the substrate 1 and the lower surface of the cover sheet 2, one end of the microchannel 3 is connected to the sample addition hole 4, and the other end is connected to the driving port 5, the water-absorbing material 6 absorbs the liquid in the microchannel 3 through the driving port 5, and the water-absorbing material 6 is a movable filter paper.

[0034] After the sample enters the microchannel 3 through the sample loading hole 4 , small particles in the sample will be retained on the block 31 a of the detection area 31 during the flow, and the remaining liquid will continue to flow to the driving port 5 and be adsorbed by the water-absorbing material 6 .

[0035] Specifically, a groove is provided on the lower surface of the cover sheet 2 along its length direction, and the cover sheet 2 and the upper surface of the substrate 1 are enclosed by the groove to form a microchannel 3.

[0036] like Figure 3 , Figure 4 As shown, there are multiple blocks 31a, and the interval between two adjacent blocks 31a is 1um-20um. The blocks 31a are one or both of "cylindrical" and "plate-shaped". Due to the different sizes of crystals and cells, different intervals can be set to retain all crystals and cells in different intervals without affecting the flow of the liquid.

[0037] like Figure 5 As shown, the height of the stopper 31a is smaller than the height of the microchannel 3, and the height of the microchannel 3 is 20um-30um.

[0038] Specifically, the sample addition hole 4 is disposed on the cover plate 2 , and the sample addition hole 4 is communicated with the microchannel 3 .

[0039] like Figure 6 As shown, the shapes of the substrate 1 and the cover sheet 2 are not limited to rectangles, but can also be other shapes, such as circles. The microchannel 3 can be made into a spiral structure accordingly, and the sample loading hole 4 is arranged at the center of the circle and connected to the microchannel 3. The driving port 5 is correspondingly arranged at the other end of the spiral structure of the microchannel 3. The liquid in the microchannel can be absorbed by the water-absorbing material 6 or the liquid in the microchannel 3 can be thrown out by a centrifugal device.

[0040] On the other hand, the present invention also provides an application of using the glass slide chip in any of the above technical solutions for detection, comprising at least the following steps:

[0041] 1) Add sample: Add the sample to be tested into the sample well to make the sample flow into the detection area, and then bring the absorbent material into contact with the drive port to absorb the liquid in the microchannel; at this time, the cells, bacteria, crystals or other tiny particles in the sample are retained on the block of the microchannel.

[0042] 2) Drying: Blow dry quickly with 37°C dry air or bake in an oven at 37°C for 30-60 minutes to dry out the moisture inside the glass slide to facilitate subsequent microscopic examination.

[0043] On the other hand, the present invention also provides an application of using the glass slide chip in any of the above technical solutions for detection, comprising at least the following steps:

[0044] 1) Adding samples: Add the sample to be tested to the sample well, allowing the sample to flow into the detection area, and then contact the water-absorbing material with the driving port to absorb the liquid in the microchannel;

[0045] 2) Elution: Add cleaning liquid drops into the sample loading hole to allow the cleaning liquid to flow into the detection area, and then contact the absorbent material with the drive port to absorb the liquid in the microchannel; clean the sample retained in the microchannel to remove impurities.

[0046] 3) Staining: Add the dye droplets into the sample well to allow the dye to flow into the detection area, and then contact the absorbent material with the drive port to absorb the liquid in the microchannel; stain the cells, bacteria, crystals or other tiny particles retained in the sample to facilitate subsequent microscopic observation.

[0047] 4) Elution again: Add cleaning solution drops into the sample loading hole to allow the cleaning solution to flow into the detection area, and then bring the absorbent material into contact with the drive port to absorb the liquid in the microchannel; wash away the excess dye in the microchannel.

[0048] 5) Drying: Blow dry quickly with 37℃ dry air or bake in an oven at 37℃ for 30-60 minutes.

[0049] Example

[0050] Enrichment of red blood cells and tiny calcified particles in urine using microfluidic glass chips

[0051] 1. Materials and Methods

[0052] 1. Several microfluidic glass chips

[0053] 2. Normal urine

[0054] 3. Plastic straws

[0055] 4. Normal human venous whole blood sample

[0056] 5. Lime (particle size 0.1mm-1mm)

[0057] 6. Sample gun and tip tube

[0058] 7. Electric heating fan

[0059] 2. Test steps:

[0060] 1. Sample preparation: Take 10 μl of whole blood and 0.1 μg of lime powder and add them to 1 ml of urine, mix well and set aside;

[0061] 2. Sample addition: Take 100 μl of the above sample and add it into the sample well of the microfluidic glass slide chip;

[0062] 3. Reaction: After the sample is stationary in the microfluidic channel for 2 minutes, it contacts the filter paper and absorbs water for about 6 minutes. After the urine in the sample hole is dried, it is placed under a microscope to observe the results;

[0063] 4. Drying: Use a low setting of an electric hot air blower to blow the microfluidic glass slide for about 5 minutes until the liquid inside disappears.

[0064] Results and discussion:

[0065] Under the microscope in step 3, it can be seen that a considerable number of red blood cells and lime particles remain in the detection area of ​​the glass chip, especially blocked at the edges of the columnar and plate-like structures, while very little liquid remains; in step 4, after the water is blown dry, the red blood cells and lime particles still remain in place. Since the size of the lime particles is similar to the size of crystals in urine, they can simulate crystals in urine.

[0066] It can be seen that the microfluidic glass chip can fix tangible substances such as cells and crystals, and can be used for subsequent staining or AI interpretation of results.

[0067] The above technical solutions only reflect the preferred technical solutions of the technical solutions of the present invention. Some changes that may be made to certain parts thereof by technicians in this technical field all reflect the principles of the present invention and fall within the protection scope of the present invention.

[0068] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0069] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and the like are only used for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, features defined as "first", "second", and the like may explicitly or implicitly include one or more of the features. In the description of the present invention, unless otherwise specified, "multiple" means two or more.

[0070] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "install", "connect", "connect", and "set" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood by specific circumstances.

Claims

1. A microfluidic pathology glass slide chip, comprising a substrate (1) and a cover sheet (2) pressed onto the substrate (1), wherein the substrate (1) and the cover sheet (2) enclose a microchannel (3), wherein a detection area (31) is provided in the middle of the microchannel (3), characterized in that: A block (31a) is provided in the detection area (31), and the block (31a) is alternately distributed on the upper surface of the substrate (1) and the lower surface of the cover sheet (2). One end of the microchannel (3) is connected to the sample addition hole (4), and the other end is connected to the drive port (5). The water-absorbing material (6) absorbs the liquid in the microchannel (3) through the drive port (5).

2. A microfluidic pathology glass slide chip according to claim 1, characterized in that: The lower surface of the cover sheet (2) is provided with a groove along its length direction, and the cover sheet (2) and the upper surface of the substrate (1) are enclosed by the groove to form a microchannel (3).

3. A microfluidic pathology glass slide chip according to claim 1, characterized in that: The number of the stoppers (31a) is plural, the interval between two adjacent stoppers (31a) is 1 um-20 um, and the stoppers (31a) are one or both of "cylindrical" and "plate-shaped".

4. A microfluidic pathology glass slide chip according to claim 1, characterized in that: The height of the stopper (31a) is smaller than the height of the microchannel (3), and the height of the microchannel (3) is 20 um-30 um.

5. A microfluidic pathology glass slide chip according to claim 1, characterized in that: The sample addition hole (4) is arranged on the cover sheet (2), and the sample addition hole (4) is connected to the microchannel (3).

6. An application of a microfluidic pathology glass slide chip, characterized in that: The detection using the glass chip according to any one of claims 1 to 5 comprises at least the following steps: 1) Adding samples: Add the sample to be tested to the sample well, allowing the sample to flow into the detection area, and then contact the water-absorbing material with the driving port to absorb the liquid in the microchannel; 2) Drying: Blow dry quickly with 37℃ dry air or bake in an oven at 37℃ for 30-60 minutes.

7. An application of a microfluidic pathology glass slide chip, characterized in that: The detection using the glass chip according to any one of claims 1 to 5 comprises at least the following steps: 1) Adding samples: Add the sample to be tested to the sample well, allowing the sample to flow into the detection area, and then contact the water-absorbing material with the driving port to absorb the liquid in the microchannel; 2) Elution: Add the cleaning solution dropwise into the sample loading well to allow the cleaning solution to flow into the detection area, and then contact the absorbent material with the drive port to absorb the liquid in the microchannel; 3) Staining: Add the dye solution to the sample well, let the dye solution flow into the detection area, and then contact the water-absorbent material with the driving port to absorb the liquid in the microchannel; 4) Elution again: Add the cleaning solution dropwise into the sample loading hole, allow the cleaning solution to flow into the detection area, and then contact the water-absorbent material with the driving port to absorb the liquid in the microchannel; Drying: Blow dry quickly with 37℃ dry air or bake in an oven at 37℃ for 30-60 minutes.

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