A system and detection method for identifying tumor stem cells and detecting their drug sensitivity

Abstract

The present invention provides a system for identifying tumor stem cells and detecting their drug sensitivity and a detection method thereof. The system includes a plurality of devices and units such as an incubator, a tissue culture unit, a reagent loading unit, a pipetting unit, a laser unit, a microscope unit, a control unit and a power supply unit. The incubator includes a door and a box body, and each functional unit is arranged in the box body, and a partition is arranged in the middle section of the box body. The tissue culture unit is arranged in the front box body, and includes a tissue culture platform composed of a plurality of tissue culture tanks, and the tumor tissue slices to be tested are placed in the tissue culture tanks in sequence. The reagent loading unit is used to load various liquid reagents such as cell culture medium, staining solution, and washing solution, and is arranged in the rear box body. The present invention can realize the efficient identification of tumor stem cells in tumor stem cell tissue slices, and quickly obtain sensitive drugs that can inhibit and eliminate the tumor stem cells.

Description

Technical Field

[0001] The present invention relates to the detection field of identifying stem cells and detecting their drug sensitivity, and in particular to a system and a detection method thereof for identifying tumor stem cells and detecting their drug sensitivity. Background Art

[0002] Tumor stem cells play an important role in the survival, proliferation, metastasis and recurrence of tumors. In essence, tumor stem cells maintain the vitality of tumor cell groups through self-renewal and unlimited proliferation, and are the basis for the occurrence and development of tumors. Because tumor stem cells can remain dormant for a long time and resist a variety of anticancer drugs, tumors often recur due to the proliferation of stem cells after conventional tumor treatments have eliminated most of the ordinary tumor cells.

[0003] If tumor stem cells can be identified and eliminated, it will be significantly beneficial to the treatment of tumors and the prevention of recurrence and metastasis. However, since the number of tumor stem cells is extremely rare and the stem cell markers of different tumors may be different, it is very difficult to identify tumor stem cells and difficult to implement targeted elimination. Therefore, it is necessary to develop a device and method that can efficiently identify tumor stem cells and help explore drugs that can successfully eliminate these cells. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a system and a detection method for identifying tumor stem cells and detecting their drug sensitivity.

[0005] The technical solution adopted by the present invention is:

[0006] A system for identifying tumor stem cells and detecting their drug sensitivity, comprising:

[0007] — an incubator, comprising a door and a body, wherein each functional unit is arranged in the body, wherein a partition is arranged in the middle of the body, and the partition divides the body into a front body and a rear body, and the door is also provided with a front door and a rear door;

[0008] — a tissue culture unit, which is used to load live tumor tissue slices and keep the cells alive, is arranged in the front box, and includes a tissue culture platform composed of a plurality of tissue culture tanks, each of which can be filled with detection reagents, and the tumor tissue slices to be tested are placed in the tissue culture tanks in sequence. The tissue culture platform is in the shape of a plate and is provided with a first transverse translation mechanism and a first longitudinal translation mechanism;

[0009] —A reagent loading unit, used for loading the detection reagent, is arranged in the rear box, comprises a plurality of liquid-carrying tubes and a tube rack, the tube rack is in an integrated form, the liquid-carrying tubes are sequentially loaded into the tube rack, and is provided with a second lateral translation mechanism;

[0010] a pipetting unit, used for sucking the reagent in the liquid-carrying tube, and injecting the reagent into the tissue culture tank or removing it from the tissue culture tank, comprising a pipette and a movable rack, the movable rack being arranged on the top of the pipette and connected to the top of the box body, the pipetting unit is also provided with a second longitudinal translation mechanism for the pipette and the movable rack to move longitudinally along the top of the box body, the partition in the box body is provided with a pipetting channel for the pipette and the movable rack to pass through, the pipette is provided with a pipette telescopic mechanism, so that the pipette port can be vertically telescopic;

[0011] - A microscope unit, comprising an inverted microscope, the objective lens of which extends from the bottom surface of the box body into the box and is placed below the tissue culture platform, and can be translated to a predetermined position through the tissue culture platform to observe the growth of the tumor tissue to be tested in each tissue culture tank and collect the observed image;

[0012] - a laser unit, used to inactivate target cells in the tumor tissue slice to be tested, including a laser, whose laser emission gun is installed on the mobile frame with the muzzle facing vertically downward;

[0013] - A control unit, including a controller, a control circuit, a communicator and a display mechanism, for controlling the start and stop, operation of each electrical device and the operation of the first lateral translation mechanism, the first longitudinal translation mechanism, the second lateral translation mechanism, the second longitudinal translation mechanism and the pipette telescopic mechanism, and for communication network connection and data transmission, as well as displaying the operation status of each unit and displaying the image collected by the microscope;

[0014] A power supply unit, including a power supply and a power supply circuit. The power supply is composed of a battery and an external power supply and is used to supply power to various electrical devices.

[0015] The first transverse translation mechanism is arranged on the front box body, including a first transverse motor and a first transverse bar. The middle section of the partition is provided with a front transverse groove. The rear edge of the first transverse bar is correspondingly connected to the front transverse groove, and its front edge is connected to the first longitudinal translation mechanism. The first transverse motor causes the first transverse bar to translate transversely along the front transverse groove, and drives the first longitudinal translation mechanism to translate transversely together; the first longitudinal translation mechanism includes a first longitudinal motor, a support platform and a first longitudinal bar. The lower edge of the first longitudinal bar is connected to the front edge of the above-mentioned first transverse bar. The support surface of the support platform is upward, and its lower part is connected to the upper edge of the first longitudinal bar. The support surface of the support platform supports The tissue culture platform is carried, and the tissue culture platform is longitudinally translated along the support platform by a first longitudinal motor; the second transverse translation mechanism is arranged in the rear box body, and a rear transverse groove is provided in the middle section of the partition, and the outer edge of the tube rack is correspondingly connected to the rear transverse groove, and the tube rack is transversely translated along the rear transverse groove by the second transverse motor; the second longitudinal translation mechanism is provided with a strip protrusion on the top surface inside the box body, and the strip protrusion passes through the partition, and a strip notch is provided in the strip protrusion, and the movable rack is longitudinally translated along the strip notch by the second longitudinal motor, driving the above-mentioned devices connected to the movable rack to reciprocate between the front box body and the rear box body.

[0016] The detection reagent includes cell culture medium, anti-tumor drug solution and immunofluorescence staining reagent.

[0017] Each tissue culture tank is provided with a liquid level sensor to help ensure that the detection reagent in the tissue culture tank is within a preset amount range.

[0018] The tissue culture platform is also connected to a cover plate, including a vertical plate and a horizontal plate. The bottom end of the vertical plate is fixedly connected to the bottom of the box body, the upper end of the vertical plate is connected to one end of the horizontal plate, and the other symmetrical end of the horizontal plate is connected to the plate groove of the tissue culture platform. The depth of the plate groove is greater than the length of the horizontal plate, and the horizontal plate can cover the tissue culture groove to prevent the volatilization of the liquid in each tissue culture groove.

[0019] The front box body also includes a water container, a first temperature sensor, a humidity sensor, a carbon dioxide supplier and a carbon dioxide concentration sensor, an air conditioner and an air sterilizer. The side wall of the front box body is also provided with a ventilation grille; the rear box body also includes a refrigerator and a second temperature sensor and an air sterilizer.

[0020] The inverted microscope is a fluorescence inverted microscope, and a light source for use by the inverted microscope is arranged on the top of the front box body above the inverted microscope.

[0021] The lower part of the incubator is connected with a box seat, the controller and the communicator are arranged in the box seat, and the rest of the inverted microscope is arranged in the box seat.

[0022] The display mechanism is one or more of a computer including a display, a smart phone and a smart tablet computer.

[0023] A method for identifying tumor stem cells and detecting their drug sensitivity comprises the following steps:

[0024] S1, under aseptic technical operation, taking a number of fresh tumor tissue slices to be tested, placing them in the tissue culture tanks respectively, adding corresponding cell culture medium into the tissue culture tanks, filling the water dish with distilled water, turning on the first temperature sensor, humidity sensor, carbon dioxide supplier and air conditioner, and maintaining the tumor tissue cells to be tested in a growth environment for culture;

[0025] S2, removing the cell culture medium in the tissue culture tank, selecting an appropriate immunofluorescence staining reagent according to the antigen to be studied, and sucking the immunofluorescence staining reagent in the liquid carrier tube through a pipette, and adding it to the tissue culture tank, collecting images through the fluorescent inverted microscope, identifying cells with different labels in the tissue, and obtaining digital images on the display;

[0026] S3, identifying suspicious tumor stem cells and their corresponding position coordinates in the digital image, irradiating the suspicious tumor stem cells with a laser to kill them, then adding culture medium to continue culturing, and observing and recording the growth of tumor tissue in the tissue slice;

[0027] S4, if the tumor cells in the tumor tissue slice to be tested gradually decrease and the tumor disappears, the eliminated cells are confirmed to be tumor stem cells, and the fluorescent markers of the tumor stem cells are recorded; otherwise, the above steps S2 and S3 are repeated until the tumor stem cells are determined;

[0028] S5, using the pipette, aspirating the anti-tumor liquid in the liquid-carrying tube into one or more anti-tumor liquid compositions, each anti-tumor liquid or anti-tumor liquid composition is added to a slice in a tissue culture tank for culture, and observing and recording the changes of the labeled tumor stem cells through the inverted microscope until a therapeutic drug that can inhibit or kill the tumor stem cells is screened out.

[0029] In the front box, the temperature inside the box is maintained at 37°C, the air conditioner adjusts and maintains the temperature, and the carbon dioxide supplier and the carbon dioxide concentration sensor maintain the carbon dioxide concentration at 5%; in the rear box, the temperature inside the box is 4-8°C, and the refrigerator adjusts and maintains the temperature.

[0030] The system and detection method have the following technical effects:

[0031] Directly culturing tumor tissue slices from patients can better simulate the growth of tumor stem cells in vivo than isolating cells and then culturing them, and the test results are more reliable;

[0032] The method of immunofluorescence staining combined with laser clearing is used to identify tumor stem cells, which can identify tumor stem cells more accurately than traditional methods;

[0033] Automated operation saves manpower, the entire process is standardized, and the results are more reliable. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 It is a schematic diagram of the front box structure of the present invention;

[0035] Figure 2 The rear box structure schematic diagram of the present invention is

[0036] Figure 3 It is a schematic diagram of the front box of the present invention;

[0037] Figure 4 It is a schematic diagram of the rear box body of the present invention.

[0038] Illustrations: 1-front box body, 2-rear box body, 3-front box door, 4-rear box door, 5-partition, 6-box seat, 7-tissue culture platform, 8-tissue culture tank, 9-vertical board, 10-horizontal board, 11-liquid-carrying tube, 12-tube rack, 13-rear transverse groove, 14-mobile rack, 15-pipette, 16-pipette channel, 17-inverted microscope, 18-objective lens, 19-laser, 20-display mechanism, 21-water dish, 22-ventilation grille, 23-refrigerator, 24-front transverse groove, 25-first horizontal bar, 26-support table, 27-first vertical bar. DETAILED DESCRIPTION

[0039] The present invention will be further described below in conjunction with the accompanying drawings.

[0040] like Figure 1 The system for identifying tumor stem cells and detecting their drug sensitivity shown in the figure includes a plurality of devices and units such as an incubator, a tissue culture unit, a reagent loading unit, a pipetting unit, a laser unit, a microscope unit, a control unit and a power supply unit. The system can realize efficient identification of tumor stem cells in tumor tissue slices and quickly obtain effective drugs that can inhibit and eliminate the tumor stem cells.

[0041] The control unit includes a controller, a control circuit, a communicator and a display mechanism (20), and is used to control the start and stop of each electrical device, the operation of the first lateral translation mechanism, the first longitudinal translation mechanism, the second lateral translation mechanism, the second longitudinal translation mechanism and the pipette extension mechanism, and to connect to a communication network and transmit data, and to display the operation of each unit and the image collected by the microscope. The controller, control circuit, communicator and the like of the control unit are all located in the box seat (6), and the display mechanism (20), control buttons and the like can be embedded in the surface of the box seat (6), or can be externally connected to a terminal device with an image display function such as a computer.

[0042] The power supply unit includes a power supply and a power supply circuit. The power supply is composed of a battery and an external power supply and is used to supply power to various electrical devices.

[0043] An incubator and a box seat (6), wherein the box seat (6) is located below the incubator. The incubator is composed of a box door and a box body, and the main functional units and functional devices of the system are all arranged in the box body. The box body is preferably in the shape of a rectangular parallelepiped or a cube, and a partition (5) is arranged horizontally in the middle of the box body, and the partition (5) divides the box body into a front box body (1) and a rear box body (2). Based on the horizontal direction of the partition (5), the box doors are also correspondingly arranged as a front box door (3) and a rear box door (4), and the front box door (3) and the rear box door (4) can be sealed and provided with a visible area made of transparent material.

[0044] The front box (1) includes a water container (21), a first temperature sensor, a humidity sensor, a carbon dioxide supplier, a carbon dioxide sensor and an air conditioner. The side wall of the front box (1) is also provided with a ventilation grille (22) to maintain the tumor tissue cells to be tested in a constant temperature and humidity environment of 37°C and 5% CO2, so that the environment in which the tumor tissue cells to be tested are located can enable them to grow normally. The rear box (2) also includes a refrigerator (23), a second temperature sensor and an air sterilizer to maintain the detection reagents in each liquid carrier tube (11) stored at an ambient temperature of 4-8°C and prevent the detection reagents from being contaminated by microorganisms.

[0045] like Figure 3The tissue culture unit shown is used to load live tumor tissue slices and culture them. It is located in the front box (1) and includes a tissue culture platform (7) composed of a plurality of tissue culture tanks (8). Cell culture medium can be added to each tissue culture tank (8). The tumor tissue slices to be tested are placed in the tissue culture tanks (8) in sequence. The tissue culture platform (7) is located in the middle of the front box (1) and is also an experimental and observation platform when the system is in use. The bottom of each tissue culture tank (8) is a transparent, colorless, non-toxic material similar to the bottom of a cell culture bottle, with a thickness of 0.1-0.2 mm. Each tissue culture tank is provided with a liquid level sensor to avoid overfilling or underfilling of the added detection reagent. At the same time, the tissue culture platform (7) is a plate-shaped platform. The first transverse translation mechanism is arranged on the front box body, and includes a first transverse motor and a first transverse bar (25). The middle section of the partition (5) is provided with a front transverse groove (24). The rear edge of the first transverse bar (25) is correspondingly connected to the front transverse groove (24), and its front edge is connected to the first longitudinal translation mechanism. The first transverse motor causes the first transverse bar (25) to translate transversely along the front transverse groove (24), and drives the first longitudinal translation mechanism to translate transversely together. The first longitudinal translation mechanism includes a first longitudinal motor, a support platform (26) and a first longitudinal bar (27). The lower edge of the first longitudinal bar (27) is connected to the front edge of the first transverse bar (25). The support surface of the support platform (26) faces upward, and its lower part is connected to the upper edge of the first longitudinal bar (27). The support surface of the support platform (26) carries the tissue culture platform (7). The tissue culture platform (7) translates longitudinally along the support platform (26) through the first longitudinal motor. The first transverse mechanism and the first longitudinal mechanism enable the tissue culture platform (7) to be translated transversely and longitudinally, so that the inverted microscope (17) can completely capture images of the tissue culture tank (8) above it.

[0046] The tissue culture platform (7) is also connected to a cover plate, including a vertical plate (9) and a horizontal plate (10). The bottom end of the vertical plate (9) is fixedly connected to the bottom of the box body, the upper end of the vertical plate (9) is connected to one end of the horizontal plate (10), and the other symmetrical end of the horizontal plate (10) is connected to the plate groove of the tissue culture platform (7). The depth of the plate groove is greater than the length of the horizontal plate (10), and the horizontal plate (10) can cover the tissue culture grooves (8) to prevent the volatilization of the liquid in each tissue culture groove (8). The lateral translation of the tissue culture platform (7) can make the tissue culture grooves (8) leave the cover of the horizontal plate (10), so as to add reagents or observe.

[0047] The first lateral translation mechanism and the control unit control the first lateral translation mechanism to enable the lateral movement. The first lateral translation mechanism is arranged on the front box (1), and includes a vertical plate (9), a horizontal plate (10) and a first lateral motor. The bottom end of the vertical plate (9) is fixedly connected to the bottom of the box, the upper end of the vertical plate (9) is connected to one end of the horizontal plate (10), and the other symmetrical end of the horizontal plate (10) is connected to the groove of the tissue culture platform (7). The tissue culture platform (7) is lateral translated along the horizontal plate (10) by the motor control. The tissue culture tank is detachable. After the tissue slice is placed in the tissue culture tank in the clean bench, it is placed back into the tissue culture platform for culture. In normal times, the displacement of the tissue culture platform allows the tissue culture tank to be hidden inside the tissue culture platform (7) to avoid bacterial contamination and liquid evaporation, while ensuring the circulation of carbon dioxide gas. When the reagent is to be added, the tissue culture platform is displaced to expose the tissue culture tank so that the reagent can be added.

[0048] like Figure 4 The reagent loading unit is used to load cell culture medium, anti-tumor drug solution, immunofluorescence staining reagent, cleaning solution for cleaning pipette, etc., as well as waste liquid after the above reagents are used. The reagent loading unit is arranged in the rear box (2), including a plurality of liquid-carrying tubes (11) and a tube rack (12), the tube rack (12) is in an integrated form, the liquid-carrying tubes (11) are sequentially placed in the tube rack (12), the tube rack (12) is moved laterally by a second transverse translation mechanism, and is controlled by a control unit. The second transverse translation mechanism is arranged in the rear box (2), and a rear transverse groove (13) is provided in the middle section of the rear box of the partition (5), the outer edge of the tube rack (12) is correspondingly connected to the rear transverse groove (13), and the tube rack (12) is laterally translated along the rear transverse groove (13) by a second transverse motor.

[0049] like Figure 3 and Figure 4The pipetting unit shown is used to suck the reagent in the liquid-carrying tube (11) and inject the reagent into the corresponding tissue culture tank (8). It includes a pipette (15) and a movable rack (14), wherein the movable rack (14) is arranged on the top of the pipette (15) and connected to the top of the box. The pipetting unit is also provided with a second longitudinal translation mechanism for the pipette (15) and the movable rack (14) to move longitudinally along the top of the box, and the partition (5) in the box is provided with a pipetting channel (16) for only the pipette (15) and the movable rack (14) and other carrying devices of the movable rack (14) to pass. The second longitudinal translation mechanism is provided with a strip protrusion on the top surface of the box, and the strip protrusion passes through the partition (5), and a strip notch is provided therein. The movable frame (14) is longitudinally translated along the strip-shaped slot by a second longitudinal motor, driving the movable frame (14) and the devices such as the pipette (15) carried by it to reciprocate between the upper part of the front box and the upper part of the rear box, and the moving position and distance are controlled by the control unit. The pipette is provided with a pipette telescopic mechanism, so that the pipette port can be vertically telescopic, and is controlled by the control unit to be inserted into the liquid-carrying tube to absorb reagents, and to reach above the tissue culture tank and reach close to the tissue culture tank to add relevant reagents.

[0050] Each reagent and liquid medicine in the reagent loading unit is moved to the top of the reagent loading unit by the pipette (15) for inhalation, and the second transverse translation mechanism simultaneously causes the reagent loading unit to be transversely translated accordingly, so that the pipette (15) can accurately probe the pipette port into the designated liquid-carrying tube (11) and control the extension and retraction of the pipette, so that the pipette can easily inhale the reagent. After inhaling the reagent or liquid medicine, the second longitudinal translation mechanism causes the pipette (15) to be translated to the tissue culture platform (7), and the first transverse translation mechanism simultaneously causes the tissue culture platform (7) to be transversely translated accordingly, so that the pipette (15) can accurately probe the pipette port to the top of the tissue culture tank (8) where the corresponding reagent or liquid medicine is to be added, and control the extension and retraction of the pipette, so that the inhaled reagent or liquid medicine is easily injected into the tissue culture tank (8).

[0051] The microscope unit comprises a fluorescent inverted microscope (17). The fluorescent inverted microscope (17) has an image acquisition function and can acquire microscopic images and transmit them to a terminal mechanism with a display function. The objective lens (18) extends from the bottom surface of the box body into the box body and is placed below the tissue culture platform (7). The growth of the tumor tissue to be tested in each tissue culture tank (8) can be observed by translating the tissue culture platform (7). The observed image can be acquired as needed, and the acquired image can be transmitted to the display mechanism (20) of the system or a connected computer. At the same time, a light source for use is provided on the top of the front box body (1) above the inverted microscope (17). The main body of the fluorescent inverted microscope (17) is located in the box seat (6). When the system is started and used, the tissue slice to be scanned needs to be stained with an immunofluorescence staining reagent, and the tissue culture tank (8) in which it is located is moved to the top of the objective lens (18) of the fluorescent inverted microscope (17) through the first transverse translation mechanism and the first longitudinal translation mechanism. Since the lower part of the tissue culture tank (8) is transparent, the objective lens (18) can directly collect images. Each cell in the tissue slice will display an image with a different fluorescent morphology. Different cells are marked by different immunofluorescence staining reagents. Whether the cells are marked or not is used as a basis for determining tumor stem cells, and provides a reference and positioning for the subsequent laser (19) to kill them.

[0052] The laser unit is used to inactivate target tumor stem cells in the tumor tissue slice to be tested. It includes a laser (19), such as a carbon dioxide laser (19), whose laser emission gun is installed on the mobile frame (14) with the muzzle facing vertically downward. When the system is started and used, the laser (19) is moved by the second longitudinal translation mechanism like the above-mentioned pipette (15), and at the same time, the first transverse translation mechanism below prompts the tissue culture platform (7) to move the corresponding tissue culture tank (8) to the top of the laser (19), and the control unit controls the accurate position of the movement of the two, so that the laser (19) can accurately kill the specified suspected tumor stem cells in the slice, and the spot size is smaller than the minimum diameter of a tumor cell, and one cell is removed each time without damaging the surrounding cells.

[0053] The system for identifying tumor stem cells and detecting their drug sensitivity, and the method for identifying tumor stem cells and detecting their drug sensitivity, comprise the following steps:

[0054] S1, taking fresh tumor tissue to be tested and freezing sections under sterile conditions, with a thickness of 5-15 microns, respectively, and placing them in the tissue culture tank (8), adding corresponding cell culture medium to the tissue culture tank (8), and filling the water container (21) with distilled water, turning on the first temperature sensor, humidity sensor, carbon dioxide supply device and air conditioner, and maintaining the cells of the tumor tissue slices to be tested growing in an environment with a concentration of 5% CO2 and a temperature of 37°C;

[0055] S2, group the tissue sections into several experimental and control groups;

[0056] S3, remove the cell culture medium in the tissue culture tank (8) in the experimental group, refer to the staining method of flow cytometry, select antibodies for tumor stem cell labeling according to literature reports or experience accumulation, suck the antibodies and other reagents labeled with fluorescent dyes in the liquid carrier tube (11) into the tissue culture tank (8) through the pipette (15), perform immunofluorescence staining on the cells in the tissue section under sterile conditions without damaging the cells, obtain digital images of the fluorescent staining of the tissue section through the fluorescent inverted microscope (17), and the same tissue section can be labeled with multiple antibodies. After the obtained photos are processed by overlapping methods, possible tumor stem cells are inferred;

[0057] S4, according to the position shown in the photo and the coordinates confirmed, use laser (19) to irradiate the suspected tumor stem cells to cause their death, then add culture medium to continue culturing, and observe the growth of tumor cells in the tissue slice. Compared with the control group, if the suspected stem cells are indeed tumor stem cells, it can be seen that the tumor growth is stagnant and all tumor cells eventually die due to lack of renewal; if the suspected stem cells are not tumor stem cells, the growth and reproduction of tumor cells in the tumor tissue slice are not affected. Repeated experiments can find clear tumor stem cells to enter the next step of the experiment.

[0058] S5, remove the ordinary culture medium of the tumor tissue slices, add culture medium containing different drugs to different tissue slices for culture, and regularly observe the growth of tumor stem cells through the above-mentioned immunofluorescence staining. Compared with the control group, if the drug has the effect of inhibiting or killing tumor stem cells, it can be seen that the tumor growth is stagnant and all tumor cells eventually die due to lack of renewal; if the drug has no effect on the growth of tumor stem cells, the growth and reproduction of tumor cells in the tumor tissue slices are not affected. Repeated experiments can find drugs that have an inhibitory or killing effect on tumor stem cells, and apply them to patients according to their conditions, thereby controlling the growth of tumors.

[0059] The above only expresses the preferred embodiments of the present invention, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that for ordinary technicians in this field, several modifications, improvements and substitutions can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached claims.

Claims

1. A system for identifying cancer stem cells and detecting their drug sensitivity, including: — an incubator, comprising a door and a body, wherein each functional unit is arranged in the body, wherein a partition is arranged in the middle of the body, and the partition divides the body into a front body and a rear body, and the door is also provided with a front door and a rear door; — a tissue culture unit, which is used to load live tumor tissue slices and keep the cells alive, is arranged in the front box, and includes a tissue culture platform composed of a plurality of tissue culture tanks, each of which can be filled with detection reagents, and the tumor tissue slices to be tested are placed in the tissue culture tanks in sequence. The tissue culture platform is in the shape of a plate and is provided with a first transverse translation mechanism and a first longitudinal translation mechanism; the detection reagents include cell culture medium, anti-tumor drug solution, and immunofluorescence staining reagent; —A reagent loading unit, used for loading the detection reagent, is arranged in the rear box, comprises a plurality of liquid-carrying tubes and a tube rack, the tube rack is in an integrated form, the liquid-carrying tubes are sequentially loaded into the tube rack, and is provided with a second lateral translation mechanism; a pipetting unit, used for sucking the reagent in the liquid-carrying tube and injecting the reagent into the tissue culture tank, comprising a pipette and a movable rack, the movable rack being arranged on the top of the pipette and connected to the top of the box body, the pipetting unit is also provided with a second longitudinal translation mechanism for the pipette and the movable rack to move longitudinally along the top of the box body, the partition in the box body is provided with a pipetting channel for the pipette and the movable rack to pass through, the pipette is provided with a pipette telescopic mechanism, so that the pipette port can be vertically telescopic; - A microscope unit, comprising an inverted microscope, the objective lens of which extends from the bottom surface of the box body into the box and is placed below the tissue culture platform, and can be translated to a predetermined position through the tissue culture platform to observe the growth of the tumor tissue to be tested in each tissue culture tank and collect the observed image; - a laser unit, used to inactivate target cells in the tumor tissue slice to be tested, including a laser, whose laser emission gun is installed on the mobile frame with the muzzle facing vertically downward; - A control unit, including a controller, a control circuit, a communicator and a display mechanism, for controlling the start and stop, operation of each electrical device and the operation of the first lateral translation mechanism, the first longitudinal translation mechanism, the second lateral translation mechanism, the second longitudinal translation and the pipette extension mechanism, and for communication network connection and data transmission, as well as displaying the operation status of each unit and displaying the image collected by the microscope; —A power supply unit, including a power supply and a power supply circuit, wherein the power supply is composed of a battery and an external power supply and is used to supply power to various electrical devices; The front box also includes a water container, a first temperature sensor, a humidity sensor, a carbon dioxide supplier and a carbon dioxide concentration sensor, an air conditioner and an air sterilizer. The side wall of the front box is also provided with a ventilation grille; the rear box also includes a refrigerator and a second temperature sensor and an air sterilizer; —The inverted microscope is a fluorescence inverted microscope, and a light source for use is provided on the top of the front box above the inverted microscope.

2. The system for identifying tumor stem cells and detecting their drug sensitivity according to claim 1, characterized in that: The first transverse translation mechanism is arranged on the front box body, and includes a first transverse motor and a first transverse bar. The middle section of the partition is provided with a front transverse groove. The rear edge of the first transverse bar is correspondingly connected to the front transverse groove, and the front edge is connected to the first longitudinal translation mechanism. The first transverse motor causes the first transverse bar to translate transversely along the front transverse groove, and drives the first longitudinal translation mechanism to translate transversely together. The first longitudinal translation mechanism comprises a first longitudinal motor, a support platform and a first longitudinal bar, wherein the lower edge of the first longitudinal bar is connected to the front edge of the first transverse bar, the support surface of the support platform faces upward, and the lower part thereof is connected to the upper edge of the first longitudinal bar, the support surface of the support platform carries the tissue culture platform, and the tissue culture platform is translated longitudinally along the support platform by the first longitudinal motor; - The second transverse translation mechanism is arranged on the rear box body, a rear transverse groove is arranged in the middle section of the partition, the outer edge of the pipe rack is correspondingly connected to the rear transverse groove, and the pipe rack is transversely translated along the rear transverse groove by the second transverse motor; -The second longitudinal translation mechanism is provided with a strip protrusion on the top surface of the box body, the strip protrusion passes through the partition, and a strip groove is provided in the strip protrusion. The movable frame is longitudinally translated along the strip groove by the second longitudinal motor, thereby driving the above-mentioned devices connected to the movable frame to reciprocate between the front box body and the rear box body.

3. The system for identifying tumor stem cells and detecting their drug sensitivity according to claim 1, characterized in that: The tissue culture platform is also connected to a cover plate, including a vertical plate and a horizontal plate. The bottom end of the vertical plate is fixedly connected to the bottom of the box body, the upper end of the vertical plate is connected to one end of the horizontal plate, and the other symmetrical end of the horizontal plate is connected to the plate groove of the tissue culture platform. The depth of the plate groove is greater than the length of the horizontal plate, and the horizontal plate can cover the tissue culture groove.

4. The system for identifying tumor stem cells and detecting their drug sensitivity according to claim 1, characterized in that: The lower part of the incubator is connected with a box seat, the controller and the communicator are arranged in the box seat, and the rest of the inverted microscope is arranged in the box seat.

5. The system for identifying tumor stem cells and detecting their drug sensitivity according to claim 1, characterized in that: The display mechanism is one or more of a computer including a display, a smart phone and a smart tablet computer.

6. A method for identifying tumor stem cells and detecting their drug sensitivity, using the system for identifying tumor stem cells and detecting their drug sensitivity according to any one of claims 1 to 5, characterized in that: The following steps are involved: S1, under aseptic technical operation, taking a number of fresh tumor tissue slices to be tested, placing them in the tissue culture tanks respectively, adding corresponding cell culture medium into the tissue culture tanks, filling the water dish with distilled water, turning on the first temperature sensor, humidity sensor, carbon dioxide supplier and air conditioner, and maintaining the tumor tissue cells to be tested in a growth environment for culture; S2, removing the cell culture medium in the tissue culture tank, selecting an appropriate immunofluorescence staining reagent according to the antigen to be studied, and sucking the immunofluorescence staining reagent in the liquid carrier tube through a pipette, and adding it to the tissue culture tank, collecting images through the fluorescent inverted microscope, identifying cells with different labels in the tissue, and obtaining digital images on the display; S3, identifying suspicious tumor stem cells and their corresponding position coordinates in the digital image, irradiating the suspicious tumor stem cells with a laser to kill them, then adding culture medium to continue culturing, and observing and recording the growth of tumor tissue in the tissue slice; S4, if the tumor cells in the tumor tissue slice to be tested gradually decrease and the tumor disappears, it is confirmed that the eliminated cells are tumor stem cells, and the fluorescent marker of the tumor stem cells is recorded; otherwise, the above steps S2 and S3 are repeated until the tumor stem cells are determined; S5, using the pipette, aspirating the anti-tumor liquid in the liquid-carrying tube into one or more anti-tumor liquid compositions, each anti-tumor liquid or anti-tumor liquid composition is added to a slice in a tissue culture tank for culture, and observing and recording the changes of the labeled tumor stem cells through the inverted microscope until a therapeutic drug that can inhibit or kill the tumor stem cells is screened out.

7. A method for identifying tumor stem cells and detecting their drug sensitivity according to claim 6, characterized in that: In the front box, the temperature inside the box is maintained at 37°C, the air conditioner adjusts and maintains the temperature, and the carbon dioxide supplier and the carbon dioxide concentration sensor maintain the carbon dioxide concentration at 5%; in the rear box, the temperature inside the box is 4-8°C, and the refrigerator adjusts and maintains the temperature.

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