Apparatus for rapid screening of tumor stem cells and method

Abstract

An apparatus for rapid screening of tumor stem cells and a method. The apparatus comprises a microscope slide (1) and a cover slip (2) for use under a microscope; the microscope slide (1) is located at the bottom and the cover slip (2) is located at the top; the microscope slide (1) and the cover slip (2) are sealed around their periphery and form an included angle of 2-10 degrees therebetween, the distance between front ends is 200-300 microns, the distance between rear ends is 100-150 microns, a liquid inlet tube (4) and a cleaning tube (5) are provided at the front ends, and a liquid outlet tube (6) is provided at the rear ends; the liquid inlet tube (4) is connected to a blood test vial (7), the cleaning tube (5) is connected to a cleaning vial or a reagent vial (8), and the liquid outlet tube (6) is connected to a blood collection vial (9); the microscope slide (1) is provided with one or more U-shaped grooves (11), the depth of the U-shaped groove (11) is 100-500 microns, and the width of the U-shaped groove (11) is 100-300 microns.

Description

A device and method capable of quickly screening tumor stem cells TECHNICAL FIELD

[0001] The present application belongs to the technical field of tumor stem cell screening, and particularly relates to a device and method capable of quickly screening tumor stem cells. BACKGROUND

[0002] Tumor stem cells are cells in tumors that have the ability to self-renew and can produce heterogeneous tumor cells. It is now clear that tumor stem cells play an important role in tumor progression and treatment resistance, most notably in blood system cancers such as leukemia. When people develop cancer, there are a small number of tumor stem cells in the blood. Therefore, in order to more accurately reflect the growth mechanism of the tumor, screening and isolating tumor stem cells from the blood is a basic work for analysis and treatment.

[0003] Tumor stem cells (CSC) are cells in tumors that have the ability to self-renew and can produce heterogeneous tumor cells, have characteristics similar to stem cells, and can self-renew and differentiate into different types of tumor cells. The tumor cells produced by tumor stem cells are heterogeneous, that is, different types of tumor cells differ in morphology, function, etc. Importantly, tumor stem cells have the ability to migrate and can migrate to other parts of the body through the blood or lymphatic system to form metastatic tumors.

[0004] Research has found that many tumors have tumor stem cells entering the blood circulation when they are 2-4 mm in size. That is, a small number of tumor stem cells will fall off to form circulating tumor cells in the early stages of tumor development, and a small number of tumor stem cells will also exist in the blood. Therefore, early screening of tumor stem cells in the blood is an effective way to detect cancer early.

[0005] For example, Chinese application No. 201610878477.9 discloses a circulating tumor cell rapid detection kit and its preparation and application method, which separates circulating tumor cells by combining immune capture and membrane filtration. The kit includes a circulating tumor cell adsorbent, a tumor cell fluorescent nanoprobes, and a white blood cell fluorescent nanoprobes. Chinese patent application No. 201910202973.6 discloses a circulating tumor cell collection device and method. Chinese patent application No. 201510151667.6 discloses a filter and an overall device for separating tumor cells in blood fluid. The above-mentioned patents filter the blood to separate circulating tumor cells in the blood and screen whether the blood contains circulating tumor cells. However, the above-mentioned devices and methods have the disadvantages of complex filtration devices, high cost, long time, and difficult operation. SUMMARY

[0006] The application provides a device capable of quickly screening tumor stem cells, to solve the technical problems of complex filtering device, high cost, long time and large operation difficulty in the prior art.

[0007] The application provides a device capable of quickly screening tumor stem cells, which comprises a glass slide and a cover glass for use under a microscope, the glass slide is located at the lower side, and the cover glass is located at the upper side, the glass slide and the cover glass are closed at the periphery and have an included angle of 2-10 degrees between them, the front end spacing is 200-300 microns, the rear end spacing is 100-150 microns, the front end is provided with a liquid inlet pipe and a cleaning pipe, and the rear end is provided with a liquid outlet pipe, the liquid inlet pipe is connected with a blood detection bottle, the cleaning pipe is connected with a cleaning bottle or a reagent bottle, and the liquid outlet pipe is connected with a blood collection bottle, and the glass slide is provided with more than one U-shaped groove, the depth of the U-shaped groove is 100-500 microns, and the width of the U-shaped groove is 100-300 microns.

[0008] As a preferred scheme, the blood detection bottle is connected with a pressure pump.

[0009] As a preferred scheme, the blood collection bottle is connected with a negative pressure pump.

[0010] As a preferred scheme, the number of the U-shaped grooves is 2-8, and the width between the U-shaped grooves is 200-800 microns.

[0011] As a preferred scheme, the depth of the U-shaped groove is 200-300 microns, and the width of the U-shaped groove is 180-250 microns.

[0012] As a preferred scheme, the material of the glass slide and the cover glass is organic glass.

[0013] The application further provides a blood screening method of the device, which adopts the following steps: collecting patient blood, entering the blood into a blood detection bottle, connecting the blood detection bottle with a liquid inlet pipe, allowing the blood to flow from the front end to the rear end, collecting the blood by a blood collection bottle, closing the liquid inlet pipe after complete collection, cleaning by a cleaning pipe or adding a detection reagent, finally, closing the cleaning pipe and the liquid outlet pipe, placing to a microscope for observation, and screening whether there is circulating tumor cell.

[0014] The application has the following beneficial effects:

[0015] 1. The application sets the spacing of the glass slide and the cover glass at two ends as different distances, so that the tumor stem cells with a larger diameter in the blood are kept on the glass slide, and most of other blood cells are discharged from the glass slide through the liquid outlet pipe.

[0016] 2、The application also adopts the mode of setting pressurization or suction on both sides of the liquid inlet pipe and the liquid outlet pipe, which significantly improves the filtering speed. Moreover, the cleaning pipe is connected with a cleaning bottle or a reagent bottle. When blood needs to be cleaned, the cleaning liquid can be added to clean the filtered blood. When the detection reagent needs to be added, the detection reagent can be added after the cleaning is completed, which is convenient for microscope observation and screening.

[0017] 3、The slide of the application adopts a transverse U-shaped groove, and the U-shaped groove is arranged to be equivalent to the size of the tumor stem cell, so that the tumor stem cell can be gathered in the U-shaped groove, which is convenient for subsequent discovery of the tumor stem cell through a microscope.

[0018] 4、The device provided by the application has low cost, can be made into a one-time detection tool, and significantly reduces the operation cost. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to make the content of the application more easily and clearly understood, the application will be further described in detail below according to specific embodiments and in combination with the drawings, in which

[0020] Fig. 1 is a structural schematic diagram of the application.

[0021] Fig. 2 is a schematic diagram of the internal structure of the application.

[0022] In the drawings, the reference numerals are:

[0023] 1, slide; 11, U-shaped groove; 2, cover glass; 3, sealing tape; 4, liquid inlet pipe; 5, cleaning pipe; 6, liquid outlet pipe; 7, blood detection bottle; 8, reagent bottle; 9, blood collection bottle. DETAILED DESCRIPTION

[0024] In order to illustrate the characteristics of the application, the application will be further described below in combination with the drawings and embodiments.

[0025] Example 1:

[0026] Please refer to figure 1 and figure 2, the embodiment of the present application provides a kind of device capable of quickly screening tumor stem cell, including the slide 1 for use under microscope and cover glass 2, the slide 1 is located below, cover glass 2 is located above, the slide 1 and cover glass 2 are closed by sealing tape 3 around, the slide 1 and cover glass 2 have 10 degrees of included angle, front end spacing is 300 microns, rear end spacing is 150 microns, the sealing tape 3 of front end is provided with a liquid inlet pipe 4 and a cleaning pipe 5, and the cavity between the slide 1 and cover glass 2 is communicated, the sealing tape 3 of rear end is provided with a liquid outlet pipe 6, and the cavity between the slide 1 and cover glass 2 is communicated;The blood detection bottle 7 is connected to the liquid inlet pipe 4, the cleaning pipe 5 is connected to a reagent bottle 8, and the blood collection bottle 9 is connected to the liquid outlet pipe 6;The slide has 6 U-shaped grooves 11, the depth of U-shaped groove 11 is 300 microns, the width of U-shaped groove 11 is 300 microns, and the width between adjacent U-shaped grooves 11 is 800 microns.By setting the spacing of the slide and cover glass at both ends to different distances, the tumor stem cells in the blood are preserved on the slide, and most other blood cells are discharged from the slide through the liquid outlet pipe.And adopt transverse U-shaped groove, and the U-shaped groove is set to be equivalent to the size of tumor stem cell, so that tumor stem cell can gather in U-shaped groove, facilitate subsequent discovery of tumor stem cell by microscope.

[0027] The embodiment also uses the mode of setting pressurization or suction on both sides of liquid inlet pipe 4 and liquid outlet pipe 6, which significantly improves the filtering speed, specifically, a pressurization pump is connected to the blood detection bottle 7, and a negative pressure pump is connected to the blood collection bottle 9.

[0028] In addition, in order to increase the strength of the slide 1 and cover glass 2 under the premise of ensuring the permeability of the slide 1 and cover glass 2, the material of the slide 1 and cover glass 2 is set to organic glass.

[0029] Embodiment 2

[0030] The embodiment provides a blood screening method for the device for quickly screening tumor stem cell of embodiment 1, specifically using the following steps: collecting patient blood, entering the blood detection bottle 7, connecting the blood detection bottle 7 with the liquid inlet pipe 4;Let blood flow from front end to rear end, collect by blood collection bottle 9, after complete collection, close the liquid inlet pipe 4, clean or add detection reagent by cleaning pipe 5, finally, close the cleaning pipe 5 and liquid outlet pipe 6, place to microscope for observation, screen whether there is circulating tumor cell.

[0031] The cells in general blood are spherical, with a diameter of 10-20 μm, while the circulating tumor stem cells are generally round or oval, composed of dozens of cells, with strong refraction, and a diameter of about 200 μm, larger than normal red blood cells and white blood cells, so that the tumor stem cells can be quickly screened out through different inlet and outlet spacings on two sides, and the detection efficiency is significantly improved.

[0032] The above examples and drawings are only used to illustrate the technical solutions of the present application, and are not a limitation of the present application. The present application is described in detail with reference to the preferred embodiments, and those skilled in the art should understand that the changes, modifications, additions or substitutions made by those skilled in the art within the essential scope of the present application do not deviate from the purpose of the present application, and should also belong to the protection scope of the claims of the present application. Other related technical structures not fully disclosed in the present application are the existing technologies in the art.

Claims

1. A device capable of rapid screening of tumor stem cells, comprising a glass slide and a cover glass for use under a microscope, the glass slide being located below and the cover glass being located above, characterized in that: The slide and cover glass are enclosed around and have an included angle of 2-10 degrees, the front end spacing is 200-300 microns, the rear end spacing is 100-150 microns, the front end has a liquid inlet pipe and a cleaning pipe, the rear end has a liquid outlet pipe; the blood detection bottle is connected to the liquid inlet pipe, the cleaning pipe is connected to a cleaning bottle or a reagent bottle, and the liquid outlet pipe is connected to a blood collection bottle; the slide has more than one U-shaped groove, the depth of the U-shaped groove is 100-500 microns, and the width of the U-shaped groove is 100-300 microns.

2. The device capable of rapidly screening tumor stem cells according to claim 1, characterized in that: The blood detection bottle is connected to a pressure pump.

3. The device capable of rapidly screening tumor stem cells according to claim 1, characterized in that: The blood collection bottle is connected to a negative pressure pump.

4. The device capable of rapidly screening tumor stem cells according to claim 1, characterized in that: The number of U-shaped grooves is 2-8, and the width between U-shaped grooves is 200-800 microns.

5. The device capable of rapidly screening tumor stem cells according to claim 1, characterized in that: The depth of the U-shaped groove is 200-300 microns, and the width of the U-shaped groove is 180-250 microns.

6. The device capable of rapidly screening tumor stem cells according to claim 1, characterized in that: The material of the slide and cover glass is organic glass.

7. A method of blood screening using the device of claim 1, characterized by The following steps are adopted: collecting patient blood, entering the blood detection bottle, connecting the blood detection bottle with the liquid inlet pipe; letting the blood flow from the front end to the rear end, collecting by the blood collection bottle, after complete collection, closing the liquid inlet pipe, cleaning or adding detection reagent by the cleaning pipe, finally, closing the cleaning pipe and the liquid outlet pipe, placing to the microscope for observation, screening whether there is circulating tumor cell.

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