High-efficiency CTC enriching and dyeing integrated equipment

Abstract

The invention relates to the technical field of biological medicine, in particular to high-efficiency CTC enrichment and dyeing integrated equipment, which comprises a heating tank, a micro-fluidic device, a liquid level sensor, a liquid level sensor, a liquid level sensor, a liquid level sensor, a liquid level sensor, a liquid level sensor, a liquid level sensor and a liquid level sensor, the liquid suction assembly is used for sucking a reagent from the test tube and adding the reagent into the microfluidic device in the heating tank; the moving assembly is arranged on the mounting frame and used for driving the liquid suction assembly to move in the horizontal direction and the vertical direction; the liquid suction assembly comprises a limiting frame and a plurality of liquid suction devices installed in the limiting frame, a movable plate is arranged at the lower end of the limiting frame, the movable plate is connected with the limiting frame through a reset spring, and the lower ends of the liquid suction devices penetrate through the limiting frame and the movable plate and are connected with liquid suction heads in a sleeving mode; two limiting blocks are fixedly connected to the opposite positions of the inner side face of the mounting frame, and the movable plate is located below the limiting blocks. The invention solves the problem of low efficiency of the existing CTC enrichment dyeing experiment.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a high-efficiency CTC enrichment and staining integrated device. Background Art

[0002] The pharmaceutical industry and the biomedical engineering industry are the two major pillars of the modern pharmaceutical industry. The biomedical industry is composed of the biotechnology industry and the pharmaceutical industry. Biomedical engineering is the comprehensive application of the principles and methods of life sciences and engineering sciences. It understands the structure, function and other life phenomena of the human body at multiple levels of molecules, cells, tissues, organs and even the entire human system from an engineering perspective. It is a general term for studying artificial materials, products, devices and system technologies used for disease prevention, disease treatment, human function assistance and health care.

[0003] In recent years, some emerging tumor diagnostic detection technologies have emerged in the field of biomedical engineering, such as circulating tumor DNA called liquid biopsy, and the detection method of circulating tumor cells (CTCs). According to research, the presence of CTCs can be found in peripheral blood before solid tumors form, so CTC detection is very suitable for early screening and early diagnosis of malignant tumors, and CTC detection has a good effect on the prognosis of malignant tumors, disease progression monitoring, recurrence prediction, postoperative micro-lesion monitoring of malignant tumors, and the design of targeted drug therapy and monitoring of treatment effects. It is currently an advanced method for early screening and diagnosis of malignant tumors. The above method can be used for detection, which can avoid trauma to patients, and the test samples are easy to obtain. It can be widely used in immunological research detection, genetic research detection or microbial detection, and can even be used in biochip and organoid research and development. As for the detection method of circulating tumor cells, the current mainstream operation method is to enrich first and then detect. Among them, in the process of detection, the collected body fluids need to be blotted, enriched and stained, and each step requires multiple additions of reagents, and there are many types of reagents. Currently, existing reagents on the market are manually taken and placed, which is inefficient. At the same time, there is a lack of reagent insulation equipment during the experiment, which leads to low reaction activity of the reagents. Both of these greatly reduce the efficiency of reagent identification. Summary of the invention

[0004] In order to solve the above-mentioned problems of the prior art, the present invention provides a high-efficiency CTC enrichment and staining integrated device, which solves the problem of low efficiency of current CTC enrichment and staining experiments.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solutions: a high-efficiency CTC enrichment and staining integrated equipment, comprising: a heating tank, which is located in a mounting frame and is used to provide a constant temperature environment for reagents in a microfluidic device; a liquid pipetting component, which is used to absorb reagents from a test tube and add them to the microfluidic device in the heating tank; a moving component, which is arranged on the mounting frame and is used to drive the liquid pipetting component to move in the horizontal and vertical directions; the liquid pipetting component comprises a limit frame and several liquid pipettes installed inside the limit frame, a movable plate is provided at the lower end of the limit frame, the movable plate is connected to the movable plate by a reset spring, the lower end of the liquid pipette passes through the limit frame and the movable plate and is sleeved with a liquid pipette head, and two limit blocks are fixedly connected to the relative positions of the inner side surface of the mounting frame, and the movable plate is located below the limit block.

[0006] By adopting the above structural design, the liquid pipetting component is driven to move in the horizontal and vertical directions by the moving component, so that the liquid pipetting component can automatically extract and transfer reagents, thereby improving the efficiency of reagent placement and solving the problem of cumbersome and laborious manual operation and low efficiency.

[0007] Preferably, the moving assembly includes a second lead screw and a slide rail which are arranged in parallel at the upper end of the mounting frame, the second lead screw and the upper end of the slide rail are slidably connected with a cross beam, a first lead screw is fixedly connected at the middle position of the cross beam, the first lead screw is vertically arranged, and the limit frame is slidably connected to the first lead screw.

[0008] With the above structural design, the second lead screw drives the crossbeam to move, thereby driving the liquid suction component to move horizontally, and the second lead screw drives the liquid suction component to move vertically.

[0009] Preferably, the pipette includes a fixedly connected sleeve end and a connecting end from bottom to top, the connecting end passes through the limit frame and is fixedly connected with a fixing bolt, the bottom end of the sleeve end cooperates with the pipette head, and the pipette is provided with a vacuum pipe along its length direction, one end of the pipe opens at the sleeve end, and the other end of the pipe opens at the fixing bolt and is connected to the vacuum pump through a connecting pipe, and a compression spring is sleeved on the connecting end, and the two ends of the compression spring are respectively abutted against the sleeve end and the limit frame.

[0010] With the above structural design, the liquid suction component moves downward with the slider of the first lead screw, and the sleeve end gradually approaches the liquid suction head bracket below until the sleeve end is tightly fitted with the liquid suction head. At this time, the compression spring plays a buffering role. After the liquid suction head is clamped on the lower end of the liquid suction device, the slider of the first lead screw moves upward, and the liquid suction head moves upward with the liquid suction device, so that the liquid suction device automatically overlaps the liquid suction head. The motor drives the liquid suction component to move along the second lead screw to the top of the reagent box, and the slider of the first lead screw drives the liquid suction component to move downward into the reagent box. The negative pressure component is turned on to absorb the reagent. After the reagent is absorbed, the first lead screw drives the liquid suction component to move upward and reset. The motor drives the liquid suction component to move along the second lead screw to the top of the microfluidic device for liquid addition. After each liquid addition, the liquid suction component moves upward and resets.

[0011] Preferably, a pipette head holder and a medical waste box are provided in the mounting frame, wherein the pipette head holder is used for placing unused pipette heads, and the medical waste box has an opening at the upper end for storing used pipette heads.

[0012] With the above structural design, the motor drives the liquid pipetting assembly to move along the second lead screw to the top of the medical waste box. At this time, the movable plate is located below the limit block, and the slider of the first lead screw moves upward. With the cooperation of the limit block, the movable plate removes the liquid pipetting head. The liquid pipetting assembly needs to add different reagents to the microfluidic device repeatedly. After the liquid pipette discards the used liquid pipetting head, the next liquid pipetting head can be installed more quickly, which saves time, eliminates the process of manually replacing the liquid pipetting head, and increases the degree of automation of the device.

[0013] Preferably, a fixing frame is placed in the heating tank, and a plurality of through holes for fixing the microfluidic device are equidistantly provided at the upper end of the fixing frame, and a discharge pipe at the tail of the microfluidic device passes through the through hole and extends from the gap between the fixing frame and the heating tank, and a water pipe extends into the heating tank, and the other end of the water pipe is connected to a water pump to inject hot water into the heating tank.

[0014] With the above structural design, the fixing frame is placed in the heating tank for use during the experiment, so that the microfluidic device can be inserted into the through hole on the fixing frame for support and fixation when placed in the heating tank for heating, thereby increasing the stability of the placement of the microfluidic device and making it less likely for the microfluidic device to tip over. The warm water directly contacts the sleeve of the microfluidic device, and due to the large specific heat capacity of water, the heat conduction effect is better and the heating is more uniform.

[0015] Preferably, the heating tank is a hollow structure, and a heating device and a heat-insulating layer are arranged in the hollow structure.

[0016] With the above structural design, the hollow structure effectively slows down the speed at which the temperature inside the heating tank is transferred to the outside, thereby achieving a heat preservation effect.

[0017] Preferably, the heating tank is divided into an outer water bath tank and an inner water bath tank, the upper end of the inner water bath tank is fixedly connected with an epitaxial plate, the epitaxial plate is overlapped above the outer water bath tank, the heating device is installed on the outer side wall of the inner water bath tank, and the insulation layer is installed on the inner side wall of the outer water bath tank.

[0018] The above structural design is adopted to set the heating tank into a detachable outer water bath and inner water bath, which is convenient for installing the heating device and the insulation layer on the one hand, and convenient for pouring out the water in the heating tank after the experiment is completed on the other hand. The insulation layer is set in the outer water bath to help slow down the heat conduction and reduce the influence of the external temperature on the internal liquid. The inner water bath is equipped with a heating device to improve the heating effect and make the heating of the heating tank more uniform.

[0019] Preferably, a temperature sensor is installed on the inner wall of the inner water bath, and a display panel for receiving an electrical signal from the temperature sensor is provided on the outside of the heating tank.

[0020] With the above structural design, the temperature sensor can monitor the temperature inside the heating tank and display it on the display panel to remind the experimenter.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The mobile component drives the liquid suction component to move in the horizontal and vertical directions, so that the liquid suction component can automatically extract and transfer reagents, improve the efficiency of reagent placement, and solve the problem of cumbersome and laborious manual operation and low efficiency;

[0023] 2. Setting a heat preservation layer in the outer water bath helps to slow down the heat conduction and reduce the influence of the external temperature on the internal liquid. Setting a heating device in the inner water bath improves the heating effect, makes the heating of the heating tank more uniform, enhances the reaction activity of the reagent, and improves the experimental efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a structural schematic diagram of the present invention;

[0025] Figure 2 It is a schematic structural diagram of the liquid-absorbing assembly of the present invention;

[0026] Figure 3 It is a structural schematic diagram of the liquid pipette of the present invention;

[0027] Figure 4 It is a structural schematic diagram of the heating tank of the present invention;

[0028] Figure 5 It is a schematic diagram of the internal structure of the heating tank of the present invention. DETAILED DESCRIPTION

[0029] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] See also Figure 1-5 The present invention provides a technical solution: a high-efficiency CTC enrichment and staining integrated device, comprising: a heating tank 10, the heating tank 10 is located in a mounting frame 1, and is used to provide a constant temperature environment for the reagent in the microfluidic device 15; a liquid suction component 6, which is used to suck the reagent from the test tube 20 and add it to the microfluidic device 15 in the heating tank 10; a moving component 2, the moving component 2 is arranged on the mounting frame 1, and is used to drive the liquid suction component 6 to move in the horizontal and vertical directions; the mounting frame 1 is fixedly connected to a base 22 of a rectangular plate structure, and the suction component 6 is fixedly connected to a base 22 of a rectangular plate structure. The liquid assembly 6 includes a limit frame 601 and several pipettes 5 installed inside the limit frame 601. The limit frame 601 is a metal plate with a C-shaped cross section. The lower end of the limit frame 601 is provided with a movable plate 603, and the movable plate 603 is connected to the movable plate 603 through a reset spring 602. The lower end of the pipette 5 passes through the limit frame 601 and the movable plate 603 and is sleeved with a pipette head 9. Two limit blocks 7 are fixedly connected to the relative position of the inner side surface of the mounting frame 1, and the movable plate 603 is located below the limit block 7. The liquid pipette assembly 6 is driven to move in the horizontal and vertical directions by the moving assembly 2, so that the liquid pipette assembly 6 can automatically extract and transfer reagents, improve the efficiency of reagent placement, and solve the problem of cumbersome and laborious manual operation and low efficiency.

[0031] The moving assembly 2 includes a second lead screw 201 and a slide rail 202 arranged in parallel at the upper end of the mounting frame 1, a slider 1 threadedly connected to the second lead screw 201 and a slider 2 slidably connected to the slide rail 202 are fixedly connected to both ends of the crossbeam 3, a first lead screw 203 is fixedly connected to the middle position of the crossbeam 3, the first lead screw 203 is arranged vertically, and the limit frame 601 is slidably connected to the first lead screw 203. The second lead screw 201 drives the crossbeam 3 to move, thereby driving the liquid absorption assembly 6 to move horizontally, and the second lead screw 201 drives the liquid absorption assembly 6 to move vertically.

[0032] The pipette 5 includes, from bottom to top, a fixedly connected sleeve end 501 and a connecting end 502, the connecting end 502 passes through the limiting frame 601 and is fixedly connected with a fixing bolt 503, the bottom end of the sleeve end 501 cooperates with the pipette head 9, the pipette 5 is provided with a vacuum pipe along its length direction, one end of the pipe opens at the sleeve end 501, the other end of the pipe opens at the fixing bolt 503 and is connected to the vacuum pump through a connecting pipe 8, a compression spring 504 is sleeved on the connecting end 502, and the two ends of the compression spring 504 are respectively abutted against the sleeve end 501 and the limiting frame 601. As the slider of the first lead screw 203 moves downward, the sleeve end 501 gradually approaches the pipette head bracket 19 below until the sleeve end 501 is tightly fitted with the pipette head 9. At this time, the compression spring 504 plays a buffering role. After the pipette head 9 is clamped on the lower end of the pipette 5, the slider of the first lead screw 203 moves upward, and the pipette head 9 moves upward with the pipette 5, so that the pipette 5 automatically clamps the pipette head 9. The motor drives the pipette assembly 6 to move above the reagent box along the second lead screw 201, and the slider of the first lead screw 203 drives the pipette assembly 6 to move downward into the reagent box. The pipette head 9 extends into the test tube 20, and the negative pressure assembly is turned on to absorb the reagent. After the reagent is absorbed, the first lead screw 203 drives the pipette assembly 6 to move upward and reset. The motor drives the pipette assembly 6 to move along the second lead screw 201 to the top of the microfluidic device 15 to add liquid. After each addition, the pipette assembly 6 moves upward and resets.

[0033] The mounting frame 1 is provided with a pipette head holder 19 and a medical waste box 21. The pipette head holder 19 is used to place unused pipette heads 9. The upper end of the medical waste box 21 is open for storing used pipette heads 9. The motor drives the pipette assembly 6 to move along the second screw 201 to the top of the medical waste box 21. At this time, the movable plate 603 is located below the limit block 7. The slider of the first screw 203 moves upward. With the cooperation of the limit block 7, the movable plate 603 removes the pipette head 9. The pipette assembly 6 needs to repeatedly add different reagents into the microfluidic device 15. After the pipette 5 discards the used pipette head 9, it can install the next pipette head 9 more quickly, which saves time, eliminates the process of manually replacing the pipette head 9, and increases the degree of automation of the device.

[0034] A fixing frame 12 is placed in the heating tank 10, and a plurality of through holes 2001 for fixing the microfluidic device 15 are equidistantly provided on the upper end of the fixing frame 12, and a discharge pipe 16 at the tail of the microfluidic device 15 passes through the through hole 2001 and extends from the gap between the fixing frame 12 and the heating tank 10, and a water pipe 23 is inserted into the interior of the heating tank 10, and a water pump is connected to the other end of the water pipe 23 to inject hot water into the heating tank 10. During the experiment, the fixing frame 12 is placed in the heating tank 10 for use, so that the microfluidic device 15 can be inserted into the through hole 2001 on the fixing frame 12 for support and fixation when placed in the heating tank 10 for heating, thereby increasing the stability of the placement of the microfluidic device 15 and preventing the microfluidic device 15 from tipping over. The warm water directly contacts the sleeve of the microfluidic device 15, and due to the large specific heat capacity of water, the heat conduction effect is better and the heating is more uniform.

[0035] The heating tank 10 is a hollow structure, in which a heating device 18 and a heat-insulating layer 17 are arranged. The hollow structure effectively slows down the speed at which the temperature inside the heating tank 10 is transferred to the outside, thereby achieving a heat-insulating effect.

[0036] The heating tank 10 is divided into an outer water bath 101 and an inner water bath 102. The upper end of the inner water bath 102 is fixedly connected with an extension plate 1021, and the extension plate 1021 is overlapped on the top of the outer water bath 101. The heating device 18 is installed on the outer side wall of the inner water bath 102, and the insulation layer 17 is installed on the inner side wall of the outer water bath 101. The heating tank 10 is set as a detachable outer water bath 101 and inner water bath 102. On the one hand, it is convenient to install the heating device 18 and the insulation layer 17, and on the other hand, it is convenient to pour out the water in the heating tank 10 after the experiment is completed. The insulation layer 17 is set in the outer water bath 101, which helps to slow down the heat conduction and reduce the influence of the external temperature on the internal liquid. The inner water bath 102 is provided with a heating device 18 to improve the heating effect and make the heating of the heating tank 10 more uniform.

[0037] A temperature sensor 14 is installed on the inner wall of the inner water bath 102, and a display panel 13 is provided outside the heating tank 10 to receive the electrical signal of the temperature sensor 14. The temperature sensor 14 can monitor the temperature inside the heating tank 10 and display it through the display panel 13 to remind the experimenter.

[0038] Working principle: Before the liquid suction component 6 sucks the reagent each time, the liquid suction head 9 must be installed first. After one reagent is injected, the liquid suction head 9 needs to be removed and a new liquid suction head 9 needs to be installed. As the slider of the first screw 203 moves downward, the liquid suction component 6 gradually approaches the liquid suction head 9 placement rack below until the sleeve end 501 fits tightly with the liquid suction head 9. At this time, the compression spring 504 plays a buffering role. After the liquid suction head 9 is clamped on the lower end of the liquid suction device 5, the slider of the first screw 203 moves upward, and the liquid suction head 9 moves upward with the liquid suction device 5, realizing the automatic overlap operation of the liquid suction device 5 with the liquid suction head 9. The motor drives the liquid pipetting assembly 6 to move above the reagent box along the second lead screw 201, and the slider of the first lead screw 203 drives the liquid pipetting assembly 6 to move downward into the reagent box. The liquid pipetting head 9 extends into the test tube 20, and the vacuum pump is turned on to absorb the reagent. After the reagent is absorbed, the first lead screw 203 drives the liquid pipetting assembly 6 to move upward and reset. The motor drives the liquid pipetting assembly 6 to move along the second lead screw 201 to the top of the microfluidic device 15 to add liquid. After each liquid addition, the liquid pipetting assembly 6 moves upward and resets. The motor drives the liquid pipetting assembly 6 to move along the second lead screw 201 to the top of the medical waste box 21. At this time, the movable plate 603 is located below the limit block 7, and the slider of the first lead screw 203 moves upward. With the cooperation of the limit block 7, the movable plate 603 removes the liquid pipetting head 9. The liquid pipetting assembly 6 needs to repeatedly add different reagents into the microfluidic device 15. After the pipette 5 discards the used liquid pipetting head 9, the next liquid pipetting head 9 can be installed more quickly.

[0039] In this embodiment, the temperature is set to 37 degrees, the water pump is turned on to fill the heating tank 10 with water, and the water pump is turned off after the water filling is completed. During the experiment, the experimenter can determine the water temperature according to the display panel 13 set on the heating tank 10, and then control the temperature and keep warm by controlling the number of heating belts installed inside the heating tank 10.

[0040] The contents not described in detail in this specification belong to the prior art known to the professional and technical personnel in this field. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for the technical solutions described in the aforementioned embodiments to be modified, or some of the technical features thereof to be replaced by equivalents, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A high-efficiency CTC enrichment and staining integrated device, characterized in that: include: A heating tank (10), the heating tank (10) being located in the mounting frame (1) and used to provide a constant temperature environment for the reagents in the microfluidic device (15); A liquid aspiration component (6), used for aspirating reagents from the test tube (20) and adding the reagents to the microfluidic device (15) in the heating tank (10); A moving assembly (2), the moving assembly (2) being arranged on the mounting frame (1) and being used for driving the liquid suction assembly (6) to move in horizontal and vertical directions; The liquid pipetting assembly (6) comprises a limit frame (601) and a plurality of liquid pipettes (5) installed inside the limit frame (601); a movable plate (603) is arranged at the lower end of the limit frame (601); the movable plate (603) is connected to the movable plate (603) via a reset spring (602); the lower end of the liquid pipette (5) passes through the limit frame (601) and the movable plate (603) and is sleeved with a liquid pipette head (9); two limit blocks (7) are fixedly connected at relative positions on the inner side surface of the mounting frame (1); and the movable plate (603) is located below the limit block (7).

2. A high-efficiency CTC enrichment and staining integrated device according to claim 1, characterized in that: The moving assembly (2) comprises a second lead screw (201) and a slide rail (202) which are arranged in parallel at the upper end of the mounting frame (1); the second lead screw (201) and the upper ends of the slide rail (202) are slidably connected to a crossbeam (3); a first lead screw (203) is fixedly connected to the middle position of the crossbeam (3); the first lead screw (203) is arranged vertically, and the limit frame (601) is slidably connected to the first lead screw (203).

3. A high-efficiency CTC enrichment and staining integrated device according to claim 1, characterized in that: The pipette (5) comprises, from bottom to top, a fixedly connected sleeve end (501) and a connecting end (502); the connecting end (502) passes through the limiting frame (601) and is fixedly connected to a fixing bolt (503); the bottom end of the sleeve end (501) cooperates with the pipette head (9); a vacuum pipe is arranged along the length direction of the pipette (5); one end of the pipe opens at the sleeve end (501); the other end of the pipe opens at the fixing bolt (503) and is connected to the vacuum pump through a connecting pipe (8); a compression spring (504) is sleeved on the connecting end (502); the two ends of the compression spring (504) are respectively in contact with the sleeve end (501) and the limiting frame (601).

4. A high-efficiency CTC enrichment and staining integrated device according to claim 1, characterized in that: The mounting frame (1) is provided with a pipette head support (19) and a medical waste box (21); the pipette head support (19) is used to place unused pipette heads (9); the medical waste box (21) has an opening at the upper end and is used to store used pipette heads (9).

5. The high-efficiency CTC enrichment and staining integrated device according to claim 1, characterized in that: A fixing frame (12) is placed in the heating tank (10), and a plurality of through holes (2001) for fixing the microfluidic device (15) are equidistantly provided at the upper end of the fixing frame (12), and a discharge pipe (16) at the tail of the microfluidic device (15) passes through the through hole (2001) and extends out from the gap between the fixing frame (12) and the heating tank (10), and a water pipe (23) extends into the interior of the heating tank (10), and the other end of the water pipe (23) is connected to a water pump for injecting hot water into the heating tank (10).

6. A high-efficiency CTC enrichment and staining integrated device according to claim 1 or 5, characterized in that: The heating tank (10) is a hollow structure, in which a heating device (18) and a heat-insulating layer (17) are arranged.

7. A high-efficiency CTC enrichment and staining integrated device according to claim 6, characterized in that: The heating tank (10) is divided into an outer water bath tank (101) and an inner water bath tank (102); an extension plate (1021) is fixedly connected to the upper end of the inner water bath tank (102); the extension plate (1021) is overlapped on the upper part of the outer water bath tank (101); the heating device (18) is installed on the outer side wall of the inner water bath tank (102); and the insulation layer (17) is installed on the inner side wall of the outer water bath tank (101).

8. A high-efficiency CTC enrichment and staining integrated device according to claim 7, characterized in that: A temperature sensor (14) is installed on the inner wall of the inner water bath (102), and a display panel (13) for receiving an electrical signal from the temperature sensor (14) is provided on the outside of the heating tank (10).

Images