CTC enrichment dyeing method and equipment

Abstract

The invention relates to the technical field of medical inspection equipment, in particular to a CTC enrichment dyeing method and equipment. The method comprises the following steps: fixing a plurality of test tubes filled with enriched staining reagents on a movable test tube rack, placing a plurality of micro-fluidic chips on a movable chip assembly bracket according to the distance between the test tubes on the test tube rack, and arranging a heating platform below the chip assembly bracket; moving the test tube rack to a position above the chip assembly bracket, dumping the test tube rack, and simultaneously scattering the reagents in the plurality of test tubes on the plurality of micro-fluidic chips in a one-to-one correspondence manner; horizontally moving the chip assembly bracket, and blocking the liquid absorption assembly below the micro-fluidic chip by utilizing the edge of the heating platform, so that the liquid absorption assembly falls off from the micro-fluidic chip; and the micro-fluidic chip separated from the liquid absorption assembly is placed on a heating platform to be dried. The method is high in cell enrichment and dyeing operation efficiency, is especially suitable for cell enrichment and dyeing operation of batch CTC chips, and improves the CTC detection efficiency.

Description

Technical Field

[0001] The present invention relates to the technical field of medical testing equipment, and in particular to a CTC enrichment and staining method and equipment. Background Art

[0002] CTC, or circulating tumor cell detection, is an emerging tumor diagnostic detection technology that has emerged in recent years. According to research, circulating tumor cells 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 and treatment effect monitoring of targeted drug therapy. It is currently an advanced method for early screening and diagnosis of malignant tumors. However, due to the low content of circulating tumor cells in peripheral blood, the capture and identification of circulating tumor cells in CTC detection is a great challenge. Another Chinese patent of the applicant, 2019103326886, discloses "Integrated reaction device and method for nuclear cell blotting enrichment and blotting staining in body fluids", with authorization announcement number CN111855333B. The reaction device includes a microporous membrane, a fixing member is provided on the fixing member, and a reaction tank is provided on the fixing member; an upper cover is provided on the reaction tank; a liftable loading platform is provided on one side of the fixing member, and a storage bin is provided under the fixing member; the storage bin is filled with a water-absorbing substance. When the cell suspension passes through the microporous membrane, cells larger than the membrane pore size are retained in the reaction vessel and adsorbed on the membrane surface due to the use of suction filtration and interception. Since the water-absorbing substance has a stable attraction speed, a gentle attraction force, and is slow and basically pressure-free, the cells adhere to the surface of the membrane. The capture and enrichment of malignant cells and pathogens in body fluids is achieved. The microporous membrane in the device is also called a microfluidic chip, which is a key component for enriching circulating tumor cells. Existing microporous membranes all use filter membranes or filter papers made of flexible fiber materials as filter materials. For example, another Chinese patent 202321916277X of the applicant discloses "A Microfluidic Device and Cell Enrichment and Staining Integrated Device", and the authorization announcement number is CN220371064U, in which the microfluidic device includes: an effusion liner, a filter assembly, and a liner net. The interior of the effusion liner is a funnel-shaped structure, and a guide strip protruding from the surface of the funnel-shaped structure is provided to guide the filtrate; the filter assembly is a microfiltration membrane provided above the effusion liner, and is used to filter the liquid sample. During the CTC detection process, blood samples and test reagents need to be added to the microfluidic device one by one, which is cumbersome and inefficient. Summary of the invention

[0003] In order to solve the above-mentioned problems of the prior art, the present invention provides a CTC enrichment staining method and equipment for cell enrichment staining operations of batch CTC chips to improve the CTC detection efficiency.

[0004] To achieve the above object, the present invention provides the following technical solutions:

[0005] The CTC enrichment and staining method described in the present invention comprises the following steps:

[0006] Step 1) fixing a plurality of test tubes containing blood samples or cell enrichment-related reagents on a movable test tube rack, placing a plurality of microfluidic chips on a movable chip assembly bracket according to the spacing between the test tubes on the test tube rack, and setting a heating platform below the chip assembly bracket;

[0007] Step 2) moving the test tube rack to the top of the chip assembly bracket, tipping the test tube rack and simultaneously dropping the reagents in the multiple test tubes onto the multiple microfluidic chips one by one;

[0008] Step 3) horizontally moving the chip component bracket, using the edge of the heating platform to block the liquid absorption component under the microfluidic chip, so that the liquid absorption component falls off the microfluidic chip;

[0009] Step 4) Move the microfluidic chip separated from the liquid aspiration component to a heating platform, and fix multiple test tubes containing cell staining related reagents on a movable test tube rack, repeat step 2), and heat the microfluidic chip through the heating platform.

[0010] Preferably, in step 2), the test tube rack is transferred to a flipping platform via a horizontal transmission device, and the flipping platform drives the test tube rack to move above the chip component bracket, and the test tube rack is tilted to simultaneously sprinkle the reagents in multiple test tubes onto multiple microfluidic chips one by one.

[0011] The above-mentioned design scheme facilitates the automated operation of dumping the test tube rack, reduces the workload of personnel, eliminates the need to use liquid pipelines to transport reagents, saves the cost of liquid pipelines and saves space.

[0012] Preferably, in step 2), multiple test tube racks are arranged in sequence on the horizontal transmission device, and one test tube rack is transferred to the flip platform at a time. The flip platform drives the test tube rack to move above the chip component bracket, and the test tube rack is tilted to simultaneously sprinkle the reagents in multiple test tubes one-to-one onto multiple microfluidic chips. Subsequently, the test tube rack and test tubes on the flip platform are cleared, and the next test tube rack on the horizontal transmission device is transferred to the flip platform, and the microfluidic chip is filled with reagents until the reagents on all test tube racks are filled.

[0013] By adopting the above-mentioned design, the filling of various reagents in the whole process of enrichment and staining can be completed automatically.

[0014] The CTC enrichment and staining device described in the present invention comprises a horizontal transmission device for horizontally moving a test tube rack, a flipping platform for flipping the test tube rack, and a slide rail platform for installing a microfluidic chip, wherein:

[0015] The horizontal transmission device is a belt conveyor;

[0016] The flip platform includes a rotating shaft arranged at the terminal end of the horizontal transmission device, a servo motor arranged at one end of the rotating shaft for driving the rotating shaft to rotate, and a test tube bracket fixedly connected to the rotating shaft, wherein the servo motor is fixedly mounted on a slider, and the slider is slidably mounted on a linear slide rail;

[0017] The slide rail platform comprises a left slide groove and a right slide groove which are arranged in parallel with each other and can be lifted and lowered simultaneously, a heating platform is arranged between the left slide groove and the right slide groove, and a second lifting device is arranged on the outer side of the heating platform.

[0018] With the above structural design, the device integrates the functions of reagent filling, enrichment and staining of CTC chips, and can simultaneously perform enrichment and staining on a group of multiple microfluidic chips to improve the efficiency of CTC detection. It can be used for cell enrichment and staining operations of batch CTC chips.

[0019] Preferably, the two linear slide rails are symmetrically arranged at the two ends of the rotating shaft, and both ends of the rotating shaft are slidably installed on the linear slide rails through sliders. The left slide groove and the right slide groove are located on the inner sides of the two linear slide rails and are parallel to the linear slide rails. The lower ends of the left slide groove and the right slide groove are provided with a first lifting device for controlling the lifting of the left slide groove and the right slide groove.

[0020] With the above structural design, the shaft is evenly stressed and runs smoothly; the left slide and the right slide can be raised and lowered, thereby improving the level of automated operation.

[0021] Preferably, a recovery mechanism is provided below the running track of the rotating shaft, and the recovery mechanism is a belt conveyor whose conveying direction is opposite to that of the translation conveying mechanism.

[0022] With the above structural design, the recovery mechanism can recover the discarded test tubes after the reagents are added.

[0023] Preferably, a chip component bracket with a rectangular frame structure is slidably installed on the left slide groove and the right slide groove, and pulleys are provided at the four corners of the chip component bracket through an extension rod perpendicular to the left slide groove or the right slide groove. The pulleys are installed in the left slide groove or the right slide groove, and the second lifting device is used for lifting the extension rod.

[0024] By adopting the above structural design, the horizontal movement and tilting action of the chip component bracket can be realized.

[0025] Preferably, a support platform for clamping the microfluidic chip is arranged inside the frame of the chip component bracket, and the support platform is a folded edge arranged on the inner side of the frame of the chip component bracket.

[0026] By adopting the above structural design, the microfluidic chip can be conveniently mounted on the chip component bracket, making it easy to take and place the microfluidic chip.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The method and device have a high degree of automation and high efficiency in cell enrichment and staining operations, and are particularly suitable for cell enrichment and staining operations of batch CTC chips, thereby improving CTC detection efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a schematic diagram of the three-dimensional structure of a CTC enrichment and staining device according to an embodiment of the present invention.

[0030] Figure 2 It is a top view structural diagram of a linear slide.

[0031] Figure 3 It is a schematic diagram of the three-dimensional structure of the chip component bracket.

[0032] Figure 4 It is a schematic diagram of the cross-sectional structure of the microfluidic chip.

[0033] Figure 5 It is a schematic diagram of the three-dimensional structure of the chip fixing part of the microfluidic chip.

[0034] Figure 6 It is a schematic diagram of the cross-sectional structure of the chip fixing part.

[0035] Figure 7 It is a schematic diagram of the three-dimensional structure of the chip body.

[0036] Figure 8 It is a schematic diagram of the three-dimensional structure of the liquid-absorbing sheet.

[0037] Fig. 9 It is a schematic diagram of the three-dimensional structure of the liquid suction component. DETAILED DESCRIPTION

[0038] 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.

[0039] The CTC enrichment and staining method described in the present invention comprises the following steps:

[0040] Step 1) fixing a plurality of test tubes containing blood samples or cell enrichment-related reagents on a movable test tube rack 8, placing a plurality of microfluidic chips 7 on a movable chip assembly bracket 6 according to the spacing between the test tubes on the test tube rack 8, and setting a heating platform 4 below the chip assembly bracket 6;

[0041] Step 2) Move the test tube rack 8 to the top of the chip assembly bracket 6, and tip the test tube rack 8 so that the reagents in multiple test tubes are simultaneously sprinkled one-to-one on multiple microfluidic chips 7; each test tube on the test tube rack 8 stores the reagents required for one filling, and the test tube rack 8 can be completely tipped over to fill all the reagents in the test tubes onto the corresponding microfluidic chip 7. Since the spacing between the test tubes corresponds to the spacing between the microfluidic chips 7, when the test tube rack 8 is completely tipped over, the reagents in each test tube are just filled onto a corresponding microfluidic chip 7.

[0042] Step 3) horizontally moving the chip component bracket 6, using the edge of the heating platform 4 to block the liquid absorption component 74 under the microfluidic chip 7, so that the liquid absorption component 74 falls off the microfluidic chip 7;

[0043] Step 4) Move the microfluidic chip 7 separated from the pipetting component 74 to the heating platform 4, and fix multiple test tubes containing cell staining related reagents on the movable test tube rack 8, repeat step 2), and heat the microfluidic chip 7 through the heating platform 4.

[0044] As a further improvement of the present invention,

[0045] In step 2), the test tube rack 8 is transferred to the flipping platform via the horizontal transmission device, and the flipping platform drives the test tube rack 8 to move above the chip assembly bracket 6, and the test tube rack 8 is tilted to simultaneously sprinkle the reagents in the multiple test tubes one-to-one onto the multiple microfluidic chips 7. In this embodiment, the flipping platform is used as a carrier for tilting the test tube rack 8, which is convenient for realizing automated mechanical operations, and can accurately control the angle of tilting the test tube rack 8 and pouring the reagents, which is conducive to improving the operating efficiency and accurately controlling the amount of reagent injection.

[0046] As a further improvement of the present invention,

[0047] In step 2), multiple test tube racks 8 are arranged on the horizontal transmission device in sequence, and one test tube rack 8 is transferred to the flip platform at a time. The flip platform drives the test tube rack 8 to move to the top of the chip assembly bracket 6, and the test tube rack 8 is tilted to simultaneously sprinkle the reagents in multiple test tubes one-to-one onto multiple microfluidic chips 7. Then, the test tube rack 8 and the test tubes on the flip platform are cleared, and the next test tube rack 8 on the horizontal transmission device 1 is transferred to the flip platform, and the microfluidic chip 7 is filled with reagents until all the reagents on the test tube racks 8 are filled. The test tube rack 8 cooperates with the horizontal transmission device 1, and multiple batches of reagent filling can be achieved using the same set of devices, thereby improving equipment utilization.

[0048] To achieve the above CTC enrichment staining method, Figure 1 As shown, the CTC enrichment and staining device described in the present invention includes a horizontal transmission device 1 for horizontally moving a test tube rack 8, a flipping platform for flipping the test tube rack 8, and a slide rail platform 5 for installing a microfluidic chip 7, wherein;

[0049] The horizontal transmission device 1 is a belt conveyor. When in use, a test tube rack 8 carrying test tubes is placed on the belt conveyor, and the belt conveyor transports the test tube racks 8 one by one to the turning platform.

[0050] The flip platform includes a rotating shaft 2 arranged at the terminal end of the horizontal transmission device 1, a servo motor 21 arranged at one end of the rotating shaft 2 for driving the rotating shaft 2 to rotate, and a test tube holder 22 fixedly connected to the rotating shaft 2. The servo motor 21 is fixedly installed on a slider, and the slider is slidably installed on a linear slide rail 10.

[0051] The test tube holder 22 is a folded plate with an L-shaped cross section. In the initial state, the bottom support plate of the test tube holder 22 is in a horizontal state and close to the terminal of the translation conveying mechanism 1. When the translation conveying mechanism 1 moves the test tube rack 8 with the test tube installed to the terminal of the translation conveying mechanism 1, it can be smoothly transferred to the bottom support plate of the test tube holder 22.

[0052] like Figure 1 , Figure 2 As shown, the servo motor 21 is fixedly mounted on the slider, and the slider is slidably mounted on the linear slide 10. The linear slide 10 includes two tracks respectively arranged at the two ends of the rotating shaft 2. The two ends of the rotating shaft 2 are respectively mounted on the two tracks through the sliders, and at least one of the sliders is mounted with the servo motor 21 for driving the rotating shaft 2 to rotate; in addition, the linear slide 10 is provided with a slider driving mechanism for driving the slider to move on the linear slide 10. Figure 2As shown, the slider driving mechanism includes a driving motor 11 fixedly mounted at one end of the linear slide 10, a lead screw 9 inserted on the slider through a thread, the front end of the lead screw 9 is connected to the end of the linear slide 10 through a bearing, and the rear end is connected to the driving motor 11 through a coupling, and the axis of the lead screw 9 is parallel to the sliding track of the linear slide 10. When the driving motor 11 is working, it drives the lead screw 9 to rotate. When the lead screw 9 rotates, the slider mounted on the lead screw 9 drives the servo motor 21 and the rotating shaft 2 to move along the axis of the lead screw 9. Of course, as other embodiments of the present invention, slider driving mechanisms with other structures can also be used, as long as the slider can be accurately driven to move on the linear slide 10.

[0053] After the test tube rack 8 is transferred to the test tube holder 22, the servo motor 21 drives the rotating shaft 2 and the test tube holder 22 to deflect a certain angle, so that the test tube rack 8 is tilted to pour the reagent in the test tube onto the microfluidic chip 7. The driving motor 11 can control the position of the rotating shaft 2 and the test tube holder 22, and move the test tube holder 22 to the top of the microfluidic chip 7 before adding the reagent.

[0054] In order to realize continuous operation, a recovery mechanism 3 is provided below the running track of the rotating shaft 2, and the recovery mechanism 3 is a belt conveyor whose conveying direction is opposite to that of the translation conveying mechanism 1. In this way, after the reagent is filled, the test tube rack 8 together with the empty test tube can be discarded to the recovery mechanism 3 by flipping the test tube bracket 22, and the recovery mechanism 3 will process them collectively.

[0055] The slide rail platform 5 includes a left slide groove 501 and a right slide groove 502 which are arranged in parallel and can be lifted and lowered simultaneously, a heating platform 4 is arranged between the left slide groove 501 and the right slide groove 502, and a second lifting device 504 is arranged outside the heating platform 4. The second lifting device 504 is two hydraulic cylinders symmetrically arranged on both sides of the heating platform 4, and can also be a screw for spiral lifting.

[0056] In addition, the left slide 501 and the right slide 502 are located on the inner side of the two linear slide rails 10 and are parallel to the linear slide rails 10. The lower ends of the left slide 501 and the right slide 502 are provided with a first lifting device 503 for controlling the lifting of the left slide 501 and the right slide 502. The first lifting device 503 is four hydraulic cylinders or screws for spiral lifting respectively arranged at the front and rear ends of the left slide 501 and the right slide 502.

[0057] The left slide groove 501 and the right slide groove 502 are slidably mounted with a chip component bracket 6 with a rectangular frame structure. A support 63 for clamping the microfluidic chip 7 is arranged inside the frame of the chip component bracket 6. The support 63 is a folded edge arranged inside the frame of the chip component bracket 6.

[0058] like Figure 1 , Figure 3 As shown, the four corners of the chip component bracket 6 are provided with pulleys 62 through extension rods 61 perpendicular to the left slide 501 or the right slide 502, and the pulleys 62 are installed in the left slide 501 or the right slide 502. When the first lifting device 503 is working, the chip component bracket 6 is driven to lift and lower in a horizontal state through the lifting and lowering of the left slide 501 and the right slide 502; the second lifting device 504 is used to lift the extension rod 61. After the chip component bracket 6 moves into place, the two extension rods 61 on the same side are just located above the second lifting device 504. When the second lifting device 504 is working, it lifts one side of the chip component bracket 6, so that one side of the chip component bracket 6 is high and the other side is low, in an inclined state. At this time, the microfluidic chip 7 on the chip component bracket 6 is also in an inclined state, which can promote the discharge of the residual reagent in the microfluidic chip 7 and reduce the residual reagent.

[0059] When in use, the test tube containing the blood sample is installed on the test tube rack 8. Multiple test tubes can be installed on the same test tube rack 8. The test tube rack 8 is placed on the translation conveying mechanism 1. The rigid chip fixing plate 72 and the reagent tank 71 are placed on the chip assembly bracket 6, so that the chip assembly bracket 6 without reagent is located between the test tube bracket 22 and the heating platform 4, and the chip body 721 is located in front of the heating platform 4. After the work starts, the driving motor 11 first moves the rotating shaft 2 and the test tube bracket 22 to the initial position, which is above the recovery mechanism 3, between the translation conveying mechanism 1 and the heating platform 4, and close to the terminal of the translation conveying mechanism 1. Subsequently, the translation conveying mechanism 1 operates and transfers the test tube rack 8 to the test tube holder 22; the driving motor 11 starts, and moves the rotating shaft 2 together with the test tube holder 22 to the top of the chip assembly holder 6. After the test tube holder 22 is in place, the servo motor 21 operates, driving the rotating shaft 2 and the test tube holder 22 to deflect a certain angle, and pouring the blood sample in the test tube into the reagent tank 71 to complete the filling of the blood sample. After the blood sample is filled, the servo motor 21 rotates in the opposite direction, discarding the test tube and the test tube rack 8 to the recovery mechanism 3, and the recovery mechanism 3 collects and processes the test tube rack 8 and the empty test tube. The servo motor 21 drives the test tube holder 22 to return to its position, and the slider driving mechanism drives the test tube holder 22 to reset, preparing for the cell enrichment reagent filling work cycle. After repeating the above steps until all the cell enrichment reagents are filled, the drive motor 11 is started to drive the test tube bracket 22 to move toward the heating platform 4. At this time, the test tube bracket 22 and the chip component bracket 6 are at the same height, and the test tube bracket 22 pushes the chip component bracket 6 to move above the heating platform 4. When the microfluidic chip 7 is pushed backward by the test tube bracket 22 along with the chip component bracket 6, the positions of the reagent tank 71 and the chip body 721 are slightly higher than the upper edge of the heating platform 4, but the lower position of the liquid suction component 74 is just blocked by the upper edge of the heating platform 4. When the reagent tank 71 moves backward with the chip component bracket 6, the liquid suction component 74 is blocked, and the claw 741 on the liquid suction component 74 slides in the claw groove 703 at the lower end of the reagent tank 71 until the claw 741 is disengaged from the claw groove 703, and the liquid suction component 74 falls off from the reagent tank 71 and falls onto the recovery mechanism 3 below, and is collected and disposed of by the recovery mechanism 3. After the chip assembly bracket 6 is moved into place, the height of the chip assembly bracket 6 can be lowered by the first lifting device 503, and it can be placed on the heating platform 4, and then the above reagent filling steps are repeated to pour the cell staining reagent into the reagent tank 71 to complete the filling of the staining reagent. The heating platform 4 heats the chip body 721 to enhance the reaction activity of the staining reagent. After the staining is completed, the first lifting device 503 raises the height of the chip assembly bracket 6 to separate it from the heating platform 4, so that the staff can take the microfluidic chip 7 conveniently.

[0060] like Figure 4As shown, the microfluidic chip 7 of the present invention is used in conjunction with a chip component bracket 6, and multiple microfluidic chips 7 can be installed on the same chip component bracket 6. The microfluidic chip 7 includes a reagent tank 71, a liquid pipetting component 74 clamped under the reagent tank 71, a rigid chip fixing plate 72 installed between the reagent tank 71 and the liquid pipetting component 74, and a chip body 721 placed in the rigid chip fixing plate 72.

[0061] like Figure 5 , Figure 6 As shown, the reagent tank 71 is a sheet structure with a liquid leakage hole 702 running through the body longitudinally, and a funnel-shaped liquid collecting groove 701 is provided on the upper surface of the reagent tank 71, and the lower end of the liquid collecting groove 701 is connected to the liquid leakage hole 702, and the upper end of the liquid collecting groove 701 is rectangular and the lower end is circular. When in use, the reagent tank 71 and the rigid chip fixing plate 72 are horizontally placed on the chip component bracket 6, and liquids such as blood samples or reagents can be collected into the liquid leakage hole 702 along the inner wall of the liquid collecting groove 701 and pass through the chip body 721.

[0062] like Figure 7 As shown, the material of the chip body 721 is preferably flexible glass, and multiple groups of long strip-shaped micropores are evenly distributed on the chip body 721. The size of the micropores allows liquids such as reagents to pass through but can intercept and capture circulating tumor cells with larger volumes. At least one diagonal line of the liquid collecting tank 701 is parallel to the side of the reagent tank 71. A drainage groove 705 is arranged on a group of diagonals of the liquid collecting tank 701. The inner end of the drainage groove 705 is connected to the liquid collecting tank 701, and the outer end is connected to the outside of the reagent tank 71. In this way, after completing cell staining, the reagent tank 71 can be tilted by raising one side of the reagent tank 71, and the excess reagent in the reagent tank 71 can be concentratedly drained to the outside of the reagent tank 71 through the drainage groove 705, and flow out along the chip component bracket 6, which is conducive to the centralized disposal of waste reagents.

[0063] In addition, if Figure 8As shown, the lower surface of the rigid chip fixing sheet 72 is provided with a liquid absorbing sheet 73, and the liquid absorbing sheet 73 includes a filter paper 731 pasted on the rigid chip fixing sheet 72, and a polymer water-absorbing material layer 732 arranged between the filter paper 731 and the chip body 721. The polymer water-absorbing material layer 732 adopts a powdered super absorbent resin, which is a new functional polymer material. It has a high water absorption function of absorbing water hundreds to thousands of times heavier than itself, and has excellent water retention performance. Once it absorbs water and swells into a hydrogel, it is difficult to separate the water even under pressure. Therefore, it has a wide range of uses in various fields such as personal hygiene products, industrial and agricultural production, and civil engineering. Super absorbent resin is generally a polymer electrolyte containing hydrophilic groups and cross-linked structures. Before absorbing water, the polymer chains are close to each other and entangled, and cross-linked to form a mesh structure, thereby achieving overall fastening. When in contact with water, water molecules penetrate into the resin through capillary action and diffusion, and the ionized groups on the chain are ionized in water. The polymer chain stretches and swells due to the electrostatic repulsion between the same ions on the chain. Due to the requirement of electrical neutrality, the counter ions cannot migrate to the outside of the resin, and the ion concentration difference between the solution inside and outside the resin forms reverse osmotic pressure. Under the action of reverse osmotic pressure, water further enters the resin to form a hydrogel, and the volume and weight will increase.

[0064] like Figure 4 As shown, when in use, the edge of the filter paper 731 is adhered to the lower surface of the rigid chip fixing sheet 72 by a water-soluble adhesive, so that the polymer water-absorbing material layer 732 faces the chip body 721. After the reagent is added, the polymer water-absorbing material layer 732 absorbs the reagent that falls through the chip body 721, and the volume expands and the weight increases, so that the liquid-absorbing sheet 73 is separated from the rigid chip fixing sheet 72.

[0065] like Fig. 9As shown, the liquid absorption assembly 74 is a container with an upper opening and receiving the leakage hole 702 on the reagent tank 71. The edge of the upper port of the liquid absorption assembly 74 is provided with a claw 741, and the claw 741 is clamped in the claw groove 703 at the lower end of the reagent tank 71. After installation, the upper end opening of the liquid absorption assembly 74 is directly opposite to the chip body 721, and the waste reagents flowing out of the leakage hole 702 and the liquid absorption sheet 73 that falls off are temporarily stored in the liquid absorption assembly 74. After the cell enrichment process is completed, since the volume and weight of the polymer water-absorbing material increase significantly after absorbing water, the liquid absorption assembly 74 is set to prevent the polymer material from overflowing after absorbing water, reducing pollution to the surrounding environment, and it is also because the volume and weight of the polymer water-absorbing material increase significantly after absorbing water, the liquid absorption sheet 73 automatically falls off from the rigid chip fixing sheet 72 under the action of its gravity and is stored in the liquid absorption assembly 74, eliminating the steps of manually peeling and collecting the liquid absorption sheet 73, reducing labor and avoiding pollution at the same time. After the filling of cell enrichment reagent is completed, when the microfluidic chip 7 is pushed backward by the test tube bracket 22 together with the chip assembly bracket 6, the position of the reagent tank 71 and the chip body 721 is slightly higher than the upper edge of the heating platform 4, but the lower-positioned liquid pipetting component 74 is just blocked by the upper edge of the heating platform 4. When the reagent tank 71 moves backward with the chip assembly bracket 6, the liquid pipetting component 74 is blocked, and the claw 741 on the liquid pipetting component 74 slides in the claw groove 703 at the lower end of the reagent tank 71 until the claw 741 disengages from the claw groove 703, and the liquid pipetting component 74 falls off from the reagent tank 71 and falls onto the recovery mechanism 3 below, and is collected and disposed of by the recovery mechanism 3.

[0066] When in use, the rigid chip fixing sheet 72 is fitted under the reagent tank 71, the liquid wicking sheet 73 is located under the chip body 721, and the liquid wicking assembly 74 is installed under the reagent tank 1. The blood sample or reagent is poured into the reagent tank 71, and after being collected in the liquid collecting tank 701, it passes through the leakage hole 702 and then passes through the chip body 721. The blood sample or reagent passing through the chip body 721 is absorbed by the liquid wicking sheet 73. The liquid wicking sheet 73 contains a polymer water-absorbing material, and its volume expands hundreds of times after absorbing water. After absorbing liquid, the liquid wicking sheet 73 is separated from the rigid chip fixing sheet 72. The liquid wicking sheet 73 that fully absorbs the reagent will eventually fall off the rigid chip fixing sheet 72 and fall into the liquid wicking assembly 74. This is the cell enrichment process. Subsequently, at least one staining reagent is poured into the reagent tank 71 respectively. After the staining of one staining reagent is completed, the extension rod 61 on one side of the reagent tank 71 is lifted by the second lifting device 504 to tilt the reagent tank 71. The reagent in the liquid collecting tank 701 will flow along the lower drainage groove 705 to the lower part of the reagent tank 71, and finally flow out along the chip component bracket 6. Then another staining reagent is poured into the reagent tank 1, and the above staining operation is repeated. The reagents in the liquid collecting tank 106 are finally collected and disposed of.

[0067] 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 CTC enrichment and staining method, characterized in that The steps include: Step 1) fixing a plurality of test tubes containing blood samples or cell enrichment-related reagents on a movable test tube rack, placing a plurality of microfluidic chips on a movable chip assembly bracket according to the spacing between the test tubes on the test tube rack, and setting a heating platform below the chip assembly bracket; Step 2) moving the test tube rack to the top of the chip assembly bracket, tipping the test tube rack and simultaneously dropping the reagents in the multiple test tubes onto the multiple microfluidic chips one by one; Step 3) horizontally moving the chip component bracket, using the edge of the heating platform to block the liquid absorption component under the microfluidic chip, so that the liquid absorption component falls off the microfluidic chip; Step 4) Move the microfluidic chip separated from the aspiration component to the heating platform, and fix multiple test tubes containing cell staining related reagents on a movable test tube rack, repeat step 2), and heat the microfluidic chip through the heating platform.

2. A CTC enrichment and staining method according to claim 1, characterized in that: In step 2), the test tube rack is transferred to the flip platform via the horizontal transmission device, and the flip platform drives the test tube rack to move above the chip component bracket, and the test tube rack is tilted to simultaneously sprinkle the reagents in multiple test tubes onto multiple microfluidic chips one by one.

3. A CTC enrichment and staining method according to claim 2, characterized in that: In step 2), multiple test tube racks are arranged on the horizontal transmission device in sequence, and one test tube rack is transferred to the flip platform at a time. The flip platform drives the test tube rack to move to the top of the chip component bracket, and the test tube rack is tilted to simultaneously sprinkle the reagents in multiple test tubes one-to-one onto multiple microfluidic chips. Then, the test tube rack and test tubes on the flip platform are cleared, and the next test tube rack on the horizontal transmission device is transferred to the flip platform, and the microfluidic chip is filled with reagents until the reagents on all test tube racks are filled.

4. A CTC enrichment and staining device, comprising a horizontal transmission device (1) for horizontally moving a test tube rack (8), a flipping platform for flipping the test tube rack (8), and a slide rail platform (5) for mounting a microfluidic chip (7), characterized in that: The horizontal transmission device (1) is a belt conveyor; The turning platform comprises a rotating shaft (2) arranged at the terminal end of the horizontal transmission device (1), a servo motor (21) arranged at one end of the rotating shaft (2) for driving the rotating shaft (2) to rotate, and a test tube holder (22) fixedly connected to the rotating shaft (2), wherein the servo motor (21) is fixedly mounted on a slide block, and the slide block is slidably mounted on a linear slide rail (10); The slide rail platform (5) comprises a left slide groove (501) and a right slide groove (502) which are arranged in parallel with each other and can be lifted and lowered simultaneously; a heating platform (4) is arranged between the left slide groove (501) and the right slide groove (502); and a second lifting device (504) is arranged on the outer side of the heating platform (4).

5. A CTC enrichment and staining device according to claim 4, characterized in that: The two linear slide rails (10) are symmetrically arranged at the two ends of the rotating shaft (2), and the two ends of the rotating shaft (2) are slidably mounted on the linear slide rails (10) through sliders. The left slide groove (501) and the right slide groove (502) are located on the inner side of the two linear slide rails (10) and are parallel to the linear slide rails (10). The lower ends of the left slide groove (501) and the right slide groove (502) are provided with a first lifting device (503) for controlling the lifting of the left slide groove (501) and the right slide groove (502).

6. A CTC enrichment and staining device according to claim 4 or 5, characterized in that: A recovery mechanism (3) is provided below the running track of the rotating shaft (2); the recovery mechanism (3) is a belt conveyor having a conveying direction opposite to that of the translation conveying mechanism (1).

7. A CTC enrichment and staining device according to claim 4 or 5, characterized in that: A chip component bracket (6) with a rectangular frame structure is slidably mounted on the left slide groove (501) and the right slide groove (502); pulleys (62) are arranged at the four corners of the chip component bracket (6) via an extension rod (61) perpendicular to the left slide groove (501) or the right slide groove (502); the pulley (62) is installed in the left slide groove (501) or the right slide groove (502); and the second lifting device (504) is used for lifting the extension rod (61).

8. A CTC enrichment and staining device according to claim 7, characterized in that: A support platform (63) for clamping the microfluidic chip (7) is arranged inside the frame of the chip component bracket (6); the support platform (63) is a folded edge arranged on the inner side of the frame of the chip component bracket (6).

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