Circulating micro-flow device for cell sorting and use method of circulating micro-flow device

Abstract

The invention discloses a circulating micro-flow device for cell sorting and a use method thereof, and belongs to the technical field of cell sorting, and the circulating micro-flow device is provided with a liquid storage tank, a filter screen plate, a reciprocating screw rod, a moving frame, a cleaning plate and a pressure spring, and positioning slots in two sides of the filter screen plate are correspondingly clamped on the surfaces of positioning clamping strips on the inner wall of the liquid storage tank; a filter screen plate is tightly attached to a cleaning plate, the cleaning plate is jacked up, so that a compression spring is extruded and deformed between a movable frame and the cleaning plate, the cleaning plate is compressed on the surface of the filter screen plate under the rebound action of the compression spring, a proper amount of cell stock solution is poured into an opening of a liquid storage tank, and meanwhile, a rotating motor is started to control a reciprocating lead screw to rotate; the movable frame reciprocates on the surface of the reciprocating screw rod to drive the cleaning plate to move back and forth on the surface of the filter screen plate, so that cell debris accumulated on the surface of the filter screen plate is cleaned, and the cell debris is prevented from being accumulated on the surface of the filter screen plate for a long time to block the filter screen plate and affect the filtering effect of the cell stock solution.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cell sorting, in particular to a circulating microfluidic device for cell sorting and a method for using the same. BACKGROUND

[0002] Cell sorting plays a crucial role in many fields such as biology, medical research, and clinical diagnosis and treatment. Accurately and efficiently separating specific types of cells from complex cell mixtures is of great significance for understanding cell function, disease mechanisms, and developing targeted treatment methods. Microfluidic devices, as important equipment for cell sorting, can achieve precise control and sorting of cells by designing specific microchannel structures and fluid driving methods. The cell stock solution flows in the flow channel, and the cells in the cell stock solution are separated according to their size under the action of inertial force when the cell stock solution flows, thereby sorting the cells in the cell stock solution according to their size.

[0003] The existing circulating microfluidic device for cell sorting has the problem that when the cell stock solution is subjected to cell sorting, a large amount of cell debris is contained in the cell stock solution, and the cell debris is easily accumulated and clogged in the filter screen during the initial filtration, thereby reducing the filtration effect of the filter screen on the cell stock solution and affecting the cell sorting effect. In addition, since the cell sorting device is generally a whole, it is difficult to replace different flow channels according to different cell sorting requirements.

[0004] Therefore, the present application provides a circulating microfluidic device for cell sorting and a method for using the same to solve the above problems. SUMMARY

[0005] The present application provides a circulating microfluidic device for cell sorting and a method for using the same to solve the above problems.

[0006] To achieve the above-mentioned purpose, the present application provides the following technical scheme: The utility model provides a circulating micro -flow device for cell sorting, including concave base, the bottom fixed mounting of installation base plate is installed to the side far from the bottom of opening, the top of installation base plate is equipped with T type clamping groove symmetrically, the top of installation base plate is equipped with processing box, the bottom of processing box is fixedly installed with T type clamping strip symmetrically, and T type clamping strip penetrates the inside of T type clamping groove and is connected with sliding, the top of concave base is fixedly installed with liquid storage tank, the top of both sides of liquid storage tank is fixedly installed with clamping mechanism respectively near processing box, the inside of liquid storage tank is equipped with cleaning mechanism, the inside of liquid storage tank is fixedly installed with filter screen plate with sliding on one side of cleaning mechanism.

[0007] As a further scheme of the utility model, the opening of the side of the concave base is slidably installed with a collection box, the surfaces of both sides of the collection box are symmetrically fixedly installed with rubber strips, and the side away from the opening of the concave base is provided with a rectangular through slot.

[0008] As a further scheme of the utility model, the bottom of the circular arc surface of the processing box is provided with a drainage port, the drainage port is clamped with a plug, the top of the surfaces of both sides of the processing box is fixedly installed with a fixed clamping block respectively, the side of the top of the processing box is rotatably installed with a magnetic cover plate I through a pin shaft, the inner wall of the processing box is fixedly installed with a spiral flow channel, and a plurality of sorting baffle strips are fixedly installed at equal intervals in the track bottom of the spiral flow channel.

[0009] As a further scheme of the utility model, the bottom end of the spiral flow channel is fixedly installed with a shunt frame, the end away from the spiral flow channel of the shunt frame penetrates the position corresponding to the rectangular through slot of the processing box, the two sides of the shunt frame are fixedly installed with a drainage frame I and a drainage frame II respectively, the drainage frame I and the drainage frame II are communicated with the two sides of the shunt frame respectively, and the ends away from the shunt frame of the drainage frame I and the drainage frame II are fixedly connected with the processing box by penetrating the processing box.

[0010] As a further scheme of the utility model, the top of the liquid storage tank is rotatably installed with a magnetic cover plate II through a pin shaft, the inside of the liquid storage tank is provided with a liquid storage groove, the inner wall of the side of the liquid storage groove opening is fixedly installed with a positioning clamping strip symmetrically, the two sides of the filter screen plate are provided with a positioning slot respectively, and the positioning slot is slidably connected with the surface of the positioning clamping strip, the top of the liquid storage tank is provided with a drainage groove away from the opening of the liquid storage groove, and the drainage groove corresponds to the opening of the top end of the spiral flow channel, the top of the liquid storage tank is fixedly installed with a circulating pump inlaid on the side of the drainage groove, the input end and the output end of the circulating pump are communicated with a conduit respectively, and the conduit is communicated with the liquid storage groove and the drainage groove respectively.

[0011] As a further scheme of the present application, the clamping mechanism comprises a concave mounting frame, one side of the concave mounting frame is rotatably mounted with a fixed clamping plate through a pin shaft, and the clamping hole of the fixed clamping plate corresponds to the position of the fixed clamping block, the concave mounting frame and the fixed clamping plate are symmetrically and fixedly installed with positioning protrusions on the side away from the clamping hole of the fixed clamping plate, and the positioning protrusions on the surfaces of the concave mounting frame and the fixed clamping plate are opposite to each other in pairs, the surfaces of the concave mounting frame and the fixed clamping plate on the side of the positioning protrusions are fixedly installed with positioning sleeves respectively, a return spring is sleeved between the two opposite positioning protrusions, and the two ends of the return spring are clamped in the positioning sleeves.

[0012] As a further scheme of the present application, the cleaning mechanism comprises a reciprocating screw rod, both ends of the reciprocating screw rod are rotatably connected to the inner wall of the liquid storage tank through bearings, a bevel gear one is fixedly installed on one end of the reciprocating screw rod and penetrates the liquid storage tank, a bevel gear two is engaged with the surface of the bevel gear one, a rotating motor is fixedly installed on one side of the bevel gear two, and the rotating motor is fixedly connected to the outer surface of the liquid storage tank, and a moving frame is threadedly connected to the surface of the reciprocating screw rod and penetrates the reciprocating screw rod.

[0013] As a further scheme of the present application, the moving frame is slidably installed with pressing rods on both sides, a limiting plate is fixedly installed on one end of the pressing rods and penetrates the moving frame, a cleaning plate is fixedly installed between the ends of the two pressing rods away from the limiting plate, a pressing spring is sleeved on the surface of the pressing rods between the moving frame and the cleaning plate, limiting slide rods are slidably installed at both ends of the moving frame and are fixedly connected to the inner wall of the liquid storage tank.

[0014] A method for using a circulating microfluidic device for cell sorting, the method comprising the steps of: According to the cell sorting requirements, a processing box with a suitable spiral flow channel is selected, the bottom of the processing box is placed on the bottom of the installation base plate, the T-shaped clamping strip on the bottom of the processing box is matched with the T-shaped clamping groove, the processing box is pushed to move on the top of the installation base plate until the T-shaped clamping strip is inserted into the corresponding T-shaped clamping groove, the processing box is gradually attached to the concave base and one side of the liquid storage tank until the top end of the spiral flow channel is attached to the inside of the liquid discharge groove, at the same time, the shunt frame is inserted into the rectangular through groove, the collection box is inserted into the opening of the concave base, the rubber strip is deformed by being squeezed on both sides of the collection box, so that the collection box is clamped and fixed in the concave base, when the processing box is gradually attached to one side of the liquid storage tank, the fixed clamping block is attached to the clamping hole of the fixed clamping plate, the fixed clamping plate is lifted to rotate and rise in the concave mounting frame, the return spring is deformed by being squeezed between the corresponding positioning protrusions, until the fixed clamping block slides through the clamping hole of the fixed clamping plate, the fixed clamping plate is reset under the rebounding action of the return spring, the clamping hole of the fixed clamping plate is clamped with the fixed clamping block, so that the processing box is clamped and fixed on one side of the liquid storage tank. Flip open the second magnetic cover plate, match the positioning slots on both sides of the filter screen plate with the positioning strips on the inner wall of the liquid storage tank, push the filter screen plate into the liquid storage tank, and slide the positioning slots on the surface of the corresponding positioning strips to ensure that the filter screen plate is tightly attached to the cleaning plate. The filter screen plate lifts up the cleaning plate, so that the cleaning plate extrusion compression spring is squeezed and deformed between the movable frame and the cleaning plate. Under the rebound action of the compression spring, the cleaning plate is pressed against the surface of the filter screen plate, and an appropriate amount of cell stock solution is poured into the liquid storage tank on one side of the filter screen plate. The cell stock solution is filtered through the filter screen plate and flows to the conduit at the input end of the circulation pump located inside the liquid storage tank. Close the second magnetic cover plate on the top of the liquid storage tank and start the circulation. The ring pump pumps the cell stock solution in the liquid reservoir into the liquid drainage tank through the catheter, and the cell stock solution then flows from the liquid drainage tank to the top of the spiral flow channel. When the cell stock solution flows in the spiral flow channel, cells of different sizes are sorted and blocked by the sorting baffles, so that cells of different sizes flow to both sides of the sorting baffles for sorting, until the sorted cell stock solution and cells flow from the spiral flow channel to the diverter rack, and are sorted by the diverter rack, liquid drainage rack 1, and liquid drainage rack 2, so that cells of different sizes are discharged from liquid drainage rack 1 and liquid drainage rack 2, and excess cell stock solution flows from the diverter rack to the collection box inside the concave base for centralized processing, until the sorting of the cell stock solution in the liquid reservoir is completed; At the same time, the filter plate filters the cell debris in the cell stock solution, so that the cell debris accumulates on one side of the filter plate, and the rotating motor is started to control the reciprocating screw to rotate synchronously under the meshing action of bevel gear one and bevel gear two, so that the movable frame moves back and forth on the surface of the reciprocating screw, and at the same time, the two ends of the movable frame slide on the surface of the corresponding limit slide rod to ensure that the movable frame does not tilt or deflect during the movement. The cleaning plate moves with the movable frame on one side of the filter plate to clean the cell debris accumulated on the surface of the filter plate, so as to prevent the cell debris from accumulating on the surface of the filter plate and affecting the filtering effect of the filter plate on the cell stock solution.

[0015] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention is provided with a liquid storage tank, a filter screen plate, a reciprocating screw rod, a movable frame, a cleaning plate and a compression spring. After the filter screen plate is inserted into the liquid storage tank, the positioning slots on both sides of the filter screen plate are correspondingly stuck on the surface of the positioning card strips on the inner wall of the liquid storage tank, the filter screen plate is tightly attached to the cleaning plate, and the cleaning plate is lifted up, so that the compression spring is squeezed and deformed between the movable frame and the cleaning plate. Under the rebound action of the compression spring, the cleaning plate is pressed against the surface of the filter screen plate, and an appropriate amount of cell stock solution is poured into the opening of the liquid storage tank. At the same time, the rotating motor is started to control the reciprocating screw rod to rotate, so that the movable frame moves back and forth on the surface of the reciprocating screw rod, driving the cleaning plate to move back and forth on the surface of the filter screen plate, and cleaning the cell debris accumulated on the surface of the filter screen plate, thereby preventing the cell debris from accumulating on the surface of the filter screen plate for a long time and clogging the filter screen plate, thereby affecting the filtering effect of the cell stock solution; 2. The application sets the installation base plate, processing box, fixed clamping block, fixed clamping plate and reset spring, the processing box is placed on the top of the installation base plate, the T-shaped clamping strip at the bottom of the processing box corresponds to the T-shaped clamping groove, the processing box is pushed to move on the top of the installation base plate until the T-shaped clamping strip is inserted into the corresponding T-shaped clamping groove, the processing box gradually approaches the liquid storage tank, the fixed clamping block on both sides of the processing box approaches the fixed clamping plate, the fixed clamping plate is lifted and rotated, the reset spring is squeezed and deformed between the corresponding positioning protrusions, until the fixed clamping block passes through the clamping hole of the fixed clamping plate, the reset spring rebounds and lifts the fixed clamping plate, the clamping hole of the fixed clamping plate is clamped with the fixed clamping block, so that the processing box is clamped and fixed on one side of the liquid storage tank, and the processing box is fixed on the top of the installation base plate, which is convenient for replacing the corresponding processing box according to the needs of cell sorting. 3. The application sets the circulating pump, the liquid discharge groove, the spiral flow channel, the sorting baffle, the shunt frame, the liquid discharge frame one and the liquid discharge frame two, the circulating pump is started to pump the cell stock solution filtered through the filter screen plate in the liquid storage tank into the liquid discharge groove, the cell stock solution in the liquid discharge groove gradually flows to the top end of the spiral flow channel, so that the cell stock solution flows in the spiral flow channel, due to the different inertia forces of cells of different sizes in the cell stock solution when flowing, cells of different sizes flow to both sides of the sorting baffle for sorting, after sorting, the cells and the cell stock solution flow to the shunt frame along the spiral flow channel, the excess cell stock solution flows into the collection box from the shunt frame for centralized treatment, and cells of different sizes are discharged from the processing box through the liquid discharge frame one and the liquid discharge frame two, so that the sorting of cells of different sizes is realized. BRIEF DESCRIPTION OF DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the application, the following will briefly introduce the drawings needed to be used in the description of the embodiments. Obviously, the drawings in the following description are only some embodiments of the application, and other drawings can also be obtained by those skilled in the art without any creative effort on the basis of these drawings.

[0017] Figure 1 It is an expanded structure diagram of the application; Figure 2 It is an expanded structure diagram of the concave base and the installation base plate of the application; Figure 3 It is a cross-sectional expanded structure diagram of the processing box and the spiral flow channel of the application; Figure 4 It is a structure diagram of the spiral flow channel and the sorting baffle of the application; Figure 5 It is an expanded structure diagram of the liquid storage tank and the cleaning mechanism of the application; Figure 6 It is a structure diagram of the clamping mechanism of the application; Figure 7It is a schematic view of sectional structure of reciprocating wire rod and moving frame of the present application. Figure 8 It is a schematic view of structure of filter screen plate of the present application.

[0018] In the drawings, the components represented by each reference numeral are listed as follows: 1, concave base; 101, collection box; 102, rubber strip; 103, rectangular through slot; 2, mounting bottom plate; 201, T-shaped clamping groove; 3, processing box; 301, T-shaped clamping strip; 302, liquid discharge port; 303, blocking block; 304, fixed clamping block; 305, magnetic cover plate one; 306, spiral flow channel; 307, sorting stopper; 308, flow distribution frame; 309, liquid discharge frame one; 310, liquid discharge frame two; 4, liquid storage tank; 401, magnetic cover plate two; 402, liquid storage tank; 403, positioning clamping strip; 404, liquid discharge tank; 405, circulating pump; 5, clamping mechanism; 501, concave mounting frame; 502, fixed clamping plate; 503, positioning protrusion; 504, positioning sleeve; 505, return spring; 6, cleaning mechanism; 601, reciprocating wire rod; 602, bevel gear one; 603, bevel gear two; 604, rotating motor; 605, moving frame; 606, pressing rod; 607, limiting plate; 608, cleaning plate; 609, pressing spring; 610, limiting sliding rod; 7, filter screen plate; 701, positioning slot. DETAILED DESCRIPTION

[0019] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application.

[0020] Please refer to Figures 1-8 The present application provides a technical solution: A circulating micro-flow device for cell sorting, comprising a concave base 1, a mounting bottom plate 2 fixedly installed at the bottom of the concave base 1 away from the outlet side, a T-shaped clamping groove 201 symmetrically provided at the top of the mounting bottom plate 2 and extending to the edge, a processing box 3 provided at the top of the mounting bottom plate 2, T-shaped clamping strips 301 fixedly installed at the bottom of the processing box 3 and slidingly connected to the inside of the T-shaped clamping groove 201; a liquid storage tank 4 fixedly installed at the top of the concave base 1, clamping mechanisms 5 fixedly installed at the top of the liquid storage tank 4 and close to the processing box 3; a cleaning mechanism 6 provided inside the liquid storage tank 4, and a filter screen plate 7 slidingly installed at one side of the cleaning mechanism 6 inside the liquid storage tank 4. During operation, a suitable processing box 3 is selected according to the needs of cell sorting, and the processing box 3 is placed on the top of the mounting base 2 and pushed so that the T-shaped clip 301 at the bottom of the processing box 3 is inserted into the corresponding T-shaped slot 201 to ensure that the processing box 3 will not be misaligned when fixed on the top of the mounting base 2, until the processing box 3 is tightly attached to one side of the liquid storage tank 4, so that the clamping mechanism 5 clamps and fixes the processing box 3, and the processing box 3 is clamped and fixed on the top of the mounting base 2. Insert the filter screen plate 7 into the liquid storage tank 4 so that the filter screen plate 7 is in close contact with the cleaning mechanism 6, and pour an appropriate amount of cell stock solution into the liquid storage tank 4. The cell stock solution filtered by the filter screen plate 7 flows to one side of the liquid storage tank 4 and is pressurized and pumped into the top of the processing box 3 for sorting. The sorted cell stock solution is discharged outward from the processing box 3, and some excess cell stock solution flows into the concave base 1 for centralized treatment. At the same time, the cleaning mechanism 6 is started to clean the cell debris accumulated on the surface of the filter screen plate 7 to prevent the cell debris from accumulating on the surface of the filter screen plate 7 and affecting the filtration of the cell stock solution by the filter screen plate 7.

[0021] As a further solution of the present invention, a collection box 101 is slidably installed through the opening on one side of the concave base 1, and rubber strips 102 are symmetrically fixedly installed on the surfaces of both sides of the collection box 101. A rectangular through groove 103 is provided on the side of the concave base 1 away from the opening; During operation, the collection box 101 is inserted into the opening of the concave base 1, so that the rubber strips 102 on both sides of the collection box 101 are squeezed and deformed with the inner wall of the concave base 1, thereby clamping and fixing the collection box 101 inside the concave base 1, and collecting and concentrating the cell stock solution dispensed from the rectangular through groove 103 to avoid leakage of excess cell stock solution during discharge. At the same time, the concentrated cell stock solution can be sorted for the second time to improve the effect of cell sorting.

[0022] As a further solution of the present invention, a drain port 302 is provided at the bottom of the arc surface of the processing box 3, a block 303 is clamped inside the drain port 302, and a magnetic cover plate 305 is installed on one side of the top of the processing box 3 through a pin shaft; a spiral flow channel 306 is fixedly installed on the inner wall of the processing box 3, and a plurality of sorting bars 307 are fixedly installed at equal intervals on the bottom of the track of the spiral flow channel 306; a diverter rack 308 is fixedly installed at the bottom end of the spiral flow channel 306, and the end of the diverter rack 308 away from the spiral flow channel 306 passes through the processing box 3 and corresponds to the position of the rectangular through groove 103. A drain rack 1 309 and a drain rack 2 310 are fixedly mounted on both sides of the diverter rack 308, and the drain rack 1 309 and the drain rack 2 310 are respectively connected to both sides of the diverter rack 308. In this embodiment, the drain rack 1 309, the drain rack 2 310 and the diverter rack 308 are U-shaped in cross-section. The ends of the drain rack 1 309 and the drain rack 2 310 away from the diverter rack 308 respectively pass through the processing box 3 and are fixedly connected to the processing box 3. When working, the cell stock solution is introduced from the top end of the spiral flow channel 306 and flows inside the spiral flow channel 306. Due to the different sizes of the cells, the cells of different sizes are guided to the two sides of the sorting baffle 307 along with the flow of the cell stock solution for sorting. The sorted cells and the cell stock solution flow to the flow distribution frame 308. The sorted cells are discharged from the liquid discharge frame one 309 and the liquid discharge frame two 310 to the processing box 3 for collection. The excess cell stock solution flows to the inside of the collection box 101 for centralized processing. At the same time, the cell stock solution is easy to splash and fall inside the processing box 3 when flowing inside the spiral flow channel 306. After the sorting is completed, the processing box 3 is removed from the top of the mounting bottom plate 2, and the plug 303 is pulled out from the inside of the liquid discharge port 302, so that the cell stock solution splashed inside the processing box 3 is poured out to avoid the splashed cell stock solution deteriorating for a long time inside the processing box 3. The cell stock solution flows inside the spiral flow channel 306. During the flow process, the sorting baffle 307 is fixed inside the spiral flow channel 306. At the same time, because the inner ring height of the spiral flow channel 306 is lower than the outer ring, the end of the sorting baffle 307 close to the outside of the spiral flow channel 306 is higher. When the cell fragments in the cell stock solution are blocked and screened by the sorting baffle 307, the cell stock solution impacts the sorting baffle 307, and the flow impact force of the cell stock solution to the two sides of the sorting baffle 307 makes the smaller cell fragments flow to the higher end of the sorting baffle 307 along with the flowing cell stock solution, and the larger cell fragments flow to the lower end of the sorting baffle 307 along with the flowing cell stock solution. When the cell stock solution impacts the sorting baffle 307 while flowing inside the spiral flow channel 306, due to the spiral structure of the spiral flow channel 306, the cell stock solution is subjected to centrifugal force. The larger cell fragments are subjected to larger centrifugal force and are thrown to the outside of the spiral flow channel 306. The smaller cell fragments are subjected to smaller centrifugal force and are distributed in the middle or inside of the spiral flow channel 306 and are blocked by the sorting baffle 307. With the impact of the cell stock solution, the smaller cell fragments flow to the inside of the spiral flow channel 306 along with the cell stock solution, so that the cell fragments of different sizes flow to the flow distribution frame 308 from the liquid discharge frame one 309 and the liquid discharge frame two 310. The flow distribution frame 308 is symmetrically provided with two flow distribution grooves, and the two flow distribution grooves are in communication with the liquid discharge frame one 309 and the liquid discharge frame two 310, so as to distribute the liquid flowing out of the spiral flow channel 306. The liquid containing larger cell fragments flows to the liquid discharge frame one 309, the liquid containing smaller cell fragments flows to the liquid discharge frame two 310, and the excess liquid is discharged from the opening of the flow distribution frame 308.

[0023] As a further scheme of the present application, one side of the top of the liquid storage tank 4 is rotatably installed with a magnetic cover plate two 401 through a pin shaft, the inside of the liquid storage tank 4 is provided with a liquid storage groove 402, the inner wall of the opening at the top of the liquid storage groove 402 is symmetrically and fixedly installed with a positioning clamping strip 403, the two sides of the filter screen plate 7 are respectively provided with positioning insertion grooves 701, and the positioning insertion grooves 701 are slidably connected to the surface of the positioning clamping strip 403, and the filter screen plate 7 divides the liquid storage groove 402 into two spaces. One side of the top of the liquid storage tank 4 away from the opening of the liquid storage groove 402 is provided with a liquid discharge groove 404, and the position of the opening at the top end of the spiral flow channel 306 corresponds to the position of the liquid discharge groove 404, and the circulating pump 405 is embeddedly and fixedly installed on one side of the top of the liquid storage tank 4 at the liquid discharge groove 404, and the input end and the output end of the circulating pump 405 are respectively communicated with the conduits, and the conduits are respectively communicated with one space of the liquid storage groove 402 and the liquid discharge groove 404; In operation, the magnetic cover plate two 401 is turned over to open the top of the liquid storage tank 4, and then the filter screen plate 7 is inserted into the inside of the liquid storage groove 402, so that the positioning insertion grooves 701 on the two sides of the filter screen plate 7 are clamped on the surface of the positioning clamping strip 403, thereby filtering the cell stock solution poured into the inside of the liquid storage groove 402, the filtered cell stock solution flows to the other side of the liquid storage groove 402, the circulating pump 405 is started, the conduit at the input end of the circulating pump 405 pumps the filtered cell stock solution in the inside of the liquid storage groove 402 into the liquid discharge groove 404, thereby guiding the filtered cell stock solution into the spiral flow channel 306 for sorting, and at the same time, the magnetic cover plate two 401 is turned over and closely attached to the top of the liquid storage tank 4, so that the magnetic cover plate two 401 is closely attached to the top of the liquid storage tank 4, thereby preventing impurities in the air on the surface from falling into the inside of the liquid storage groove 402 and causing the cell stock solution to deteriorate.

[0024] As a further scheme of the present application, the top of the two side surfaces of the processing tank 3 is respectively fixedly installed with a fixed clamping block 304; the clamping mechanism 5 comprises a concave mounting bracket 501, the concave mounting bracket 501 is rotatably installed with a fixed clamping plate 502 through a pin shaft, the clamping hole at one end of the fixed clamping plate 502 corresponds in position to the fixed clamping block 304, the concave mounting bracket 501 and the fixed clamping plate 502 are respectively and symmetrically fixedly installed with positioning protrusions 503 away from the clamping hole of the fixed clamping plate 502, the positioning protrusions 503 on the surface of the concave mounting bracket 501 and the fixed clamping plate 502 are opposite to each other in pairs, the surface of the concave mounting bracket 501 and the fixed clamping plate 502 at one side of the positioning protrusions 503 is respectively fixedly installed with a positioning sleeve 504, a return spring 505 is sleeved between the two opposite positioning protrusions 503, and the two ends of the return spring 505 are clamped in the inside of the positioning sleeve 504; When working, when the processing box 3 is pushed to be close to the liquid storage box 4, the fixed clamping block 304 is gradually attached to the fixed clamping plate 502, one end of the fixed clamping plate 502 is lifted, the reset spring 505 is extruded and deformed between the corresponding positioning protrusions 503, and the two ends of the reset spring 505 are movable in the corresponding positioning sleeves 504, so that the two ends of the reset spring 505 cannot be dislocated and offset to cause the reset spring 505 to be separated from the surface of the positioning protrusion 503, until the fixed clamping block 304 slides through the clamping opening of the fixed clamping plate 502, the reset spring 505 rebounds to lift the fixed clamping plate 502 to reset, and the fixed clamping plate 502 is clamped and fixed on the surface of the fixed clamping block 304, so that the processing box 3 is clamped and fixed on one side of the liquid storage box 4.

[0025] As a further scheme of the present application, the cleaning mechanism 6 comprises a reciprocating screw rod 601, both ends of the reciprocating screw rod 601 are rotatably connected to the inner wall of the liquid storage tank 402 through bearings, one end of the reciprocating screw rod 601 is fixedly installed with a bevel gear one 602 penetrating through the liquid storage box 4, the surface of the bevel gear one 602 is engaged with a bevel gear two 603, a rotating motor 604 is fixedly installed on one side of the bevel gear two 603, and the rotating motor 604 is fixedly connected to the outer surface of the liquid storage box 4, a moving frame 605 is threadedly connected to the surface of the reciprocating screw rod 601 penetrating through; two pressure rods 606 are slidably installed on the two sides of the moving frame 605 respectively, a limiting plate 607 is fixedly installed on one end of the pressure rod 606 penetrating through the moving frame 605, a cleaning plate 608 is fixedly installed between the two ends of the two pressure rods 606 away from the limiting plate 607, and a pressure spring 609 is sleeved on the surface of the pressure rod 606 between the moving frame 605 and the cleaning plate 608; When working, after the filter screen plate 7 is inserted into the inside of the liquid storage tank 402 and the filter screen plate 7 is attached to the cleaning plate 608, the filter screen plate 7 lifts the cleaning plate 608, the pressure spring 609 is extruded and deformed between the moving frame 605 and the cleaning plate 608, the cleaning plate 608 is pressed on the surface of the filter screen plate 7 under the rebounding action of the pressure spring 609, the rotating motor 604 is started to control the bevel gear two 603 to rotate, the reciprocating screw rod 601 is controlled to rotate synchronously under the meshing action of the bevel gear two 603 and the bevel gear one 602, the moving frame 605 is moved back and forth on the surface of the limiting slide rod 610 with the reciprocating screw rod 601, the cleaning plate 608 is moved on the surface of the filter screen plate 7, the cell fragments accumulated on one side of the filter screen plate 7 are pushed to move, and the accumulation of the cell fragments on the surface of the filter screen plate 7 is avoided to affect the filtering effect of the filter screen plate 7.

[0026] A use method of a circulating microfluidic device for cell sorting, the use method comprising the following steps: According to the cell sorting requirements, select the processing box 3 equipped with a suitable spiral flow channel 306, put the bottom of the processing box 3 on the bottom of the installation base 2, make the T-shaped card strip 301 at the bottom of the processing box 3 correspond to the T-shaped card slot 201, push the processing box 3 to move on the top of the installation base 2 until the T-shaped card strip 301 is inserted into the corresponding T-shaped card slot 201, so that the processing box 3 gradually sticks to the concave base 1 and the side of the liquid storage tank 4 until the top of the spiral flow channel 306 sticks to the inside of the drainage groove 404, and at the same time the diverter rack 308 is inserted into the rectangular through groove 103, and the collection box 101 is inserted into the opening of the concave base 1 so that the rubber strip 102 is on both sides of the collection box 101. The side is squeezed and deformed, thereby clamping and fixing the collection box 101 inside the concave base 1. When the processing box 3 gradually comes close to the side of the liquid storage tank 4, the fixed clamping block 304 is tightly attached to the fixed clamping plate 502, and the fixed clamping plate 502 is lifted up to rotate and tilt inside the concave mounting frame 501. The extrusion return spring 505 is squeezed and deformed between the corresponding positioning protrusions 503 until the fixed clamping block 304 slides through the clamping plate 502. Under the rebound action of the return spring 505, the fixed clamping plate 502 is lifted up and reset, so that the clamping plate 502 is engaged with the fixed clamping block 304, thereby clamping and fixing the processing box 3 to one side of the liquid storage tank 4. After the filter screen 7 is in the closed position, the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen 7 is in the closed position, and the filter screen The circulating pump 405 is started to pump the cell stock solution in the liquid reservoir 402 into the drainage trough 404 through the conduit. The cell stock solution then flows from the drainage trough 404 to the top of the spiral flow channel 306. When the cell stock solution flows in the spiral flow channel 306, cells of different sizes are sorted and blocked by the sorting baffle 307, so that cells of different sizes flow to both sides of the sorting baffle 307 for sorting, until the sorted cell stock solution and cells flow from the spiral flow channel 306 to the diverter rack 308. After sorting by the diverter rack 308, the drainage rack 1 309 and the drainage rack 2 310, cells of different sizes are discharged from the drainage rack 1 309 and the drainage rack 2 310. The excess cell stock solution flows from the diverter rack 308 to the collection box 101 inside the concave base 1 for centralized processing until the sorting of the cell stock solution in the liquid reservoir 402 is completed. Meanwhile, the filter screen 7 filters the cell fragments in the cell stock solution, so that the cell fragments are accumulated on one side of the filter screen 7. The rotating motor 604 controls the synchronous rotation of the reciprocating screw rod 601 under the meshing action of the bevel gear one 602 and the bevel gear two 603, so that the moving frame 605 moves back and forth on the surface of the reciprocating screw rod 601, and the two ends of the moving frame 605 slide on the surfaces of the corresponding limiting slide rods 610, so that the moving frame 605 does not tilt and deflect during the movement. The cleaning plate 608 moves with the moving frame 605 on one side of the filter screen 7, and cleans the cell fragments accumulated on the surface of the filter screen 7, so as to avoid the influence of the accumulation of the cell fragments on the surface of the filter screen 7 on the filtering effect of the filter screen 7 on the cell stock solution.

Claims

1. A circulating microfluidic device for cell sorting, comprising a concave base (1), characterized in that: A mounting base (2) is fixedly mounted on the bottom of the concave base (1) away from the opening side, a T-shaped card slot (201) is symmetrically provided on the top of the mounting base (2), a processing box (3) is provided on the top of the mounting base (2), a T-shaped card strip (301) is symmetrically fixedly mounted on the bottom of the processing box (3), and the T-shaped card strip (301) is slidably connected to the inside of the T-shaped card slot (201), a liquid storage tank (4) is fixedly mounted on the top of the concave base (1), and a clamping mechanism (5) is fixedly mounted on the top of both sides of the liquid storage tank (4) near the processing box (3), a cleaning mechanism (6) is provided inside the liquid storage tank (4), and a filter screen (7) is slidably mounted on the inside of the liquid storage tank (4) on one side of the cleaning mechanism (6).

2. A circulating microfluidic device for cell sorting according to claim 1, characterized in that: A collection box (101) is slidably mounted through an opening on one side of the concave base (1), rubber strips (102) are symmetrically fixedly mounted on the surfaces of both sides of the collection box (101), and a rectangular through groove (103) is provided on a side of the concave base (1) away from the opening.

3. A circulating microfluidic device for cell sorting according to claim 1, characterized in that: The bottom of the arc surface of the processing box (3) is provided with a drain port (302), a blocking block (303) is clamped inside the drain port (302), and fixed blocks (304) are fixedly installed on the top of the two side surfaces of the processing box (3), and a magnetic cover plate (305) is rotatably installed on one side of the top of the processing box (3) through a pin shaft. A spiral flow channel (306) is fixedly installed on the inner wall of the processing box (3), and a plurality of sorting bars (307) are fixedly installed at equal intervals on the bottom of the track of the spiral flow channel (306).

4. A circulating microfluidic device for cell sorting according to claim 3, characterized in that: A diversion rack (308) is fixedly installed at the bottom end of the spiral flow channel (306), and the end of the diversion rack (308) away from the spiral flow channel (306) passes through the processing box (3) and corresponds to the position of the rectangular through groove (103). A drainage rack 1 (309) and a drainage rack 2 (310) are fixedly installed on both sides of the diversion rack (308), and the drainage rack 1 (309) and the drainage rack 2 (310) are respectively connected to the two sides of the diversion rack (308). The ends of the drainage rack 1 (309) and the drainage rack 2 (310) away from the diversion rack (308) respectively pass through the processing box (3) and are fixedly connected to the processing box (3).

5. A circulating microfluidic device for cell sorting according to claim 4, characterized in that: A second magnetic cover plate (401) is rotatably mounted on one side of the top of the liquid storage tank (4) via a pin shaft, a liquid storage tank (402) is provided inside the liquid storage tank (4), a positioning clip (403) is symmetrically fixedly mounted on the inner wall of one side of the opening of the liquid storage tank (402), positioning slots (701) are respectively provided on both sides of the filter screen plate (7), and the positioning slots (701) penetrate and are slidably connected to the surface of the positioning clip (403), a drainage groove (404) is provided on the side of the top of the liquid storage tank (4) away from the opening of the liquid storage tank (402), and the drainage groove (404) corresponds to the position of the opening of the top of the spiral flow channel (306), a circulation pump (405) is embedded and fixedly mounted on one side of the drainage groove (404) on the top of the liquid storage tank (4), the input end and the output end of the circulation pump (405) are respectively communicated with the conduit, and the conduit is respectively communicated with the liquid storage tank (402) and the drainage groove (404).

6. The circulating microfluidic device for cell sorting according to claim 1, characterized in that: The clamping mechanism (5) comprises a concave mounting frame (501), a fixed clamping plate (502) being rotatably mounted on one side of the concave mounting frame (501) via a pin shaft, and a clamping opening of the fixed clamping plate (502) corresponds to a position of the fixed clamping block (304), and positioning protrusions (503) are symmetrically fixedly mounted on the concave mounting frame (501) and the fixed clamping plate (502) on one side away from the clamping opening of the fixed clamping plate (502), and the positioning protrusions (503) on the surfaces of the concave mounting frame (501) and the fixed clamping plate (502) are opposed to each other, and positioning sleeves (504) are fixedly mounted on the surfaces of the concave mounting frame (501) and the fixed clamping plate (502) on one side of the positioning protrusions (503), and a return spring (505) is sleeved between the two opposing positioning protrusions (503), and both ends of the return spring (505) are clamped inside the positioning sleeve (504).

7. The circulating microfluidic device for cell sorting according to claim 5, characterized in that: The cleaning mechanism (6) includes a reciprocating screw (601), both ends of which are rotatably connected to the inner wall of the liquid storage tank (402) through bearings, one end of the reciprocating screw (601) passes through the liquid storage tank (4) and is fixedly installed with a bevel gear 1 (602), the surface of the bevel gear 1 (602) is meshed with a bevel gear 2 (603), a rotating motor (604) is fixedly installed on one side of the bevel gear 2 (603), and the rotating motor (604) is fixedly connected to the outer surface of the liquid storage tank (4), and a movable frame (605) is threadedly connected to the surface of the reciprocating screw (601).

8. The circulating microfluidic device for cell sorting according to claim 7, characterized in that: A clamping rod (606) is slidably installed on both sides of the movable frame (605), one end of the clamping rod (606) passes through the movable frame (605) and is fixedly installed with a limit plate (607), a cleaning plate (608) is fixedly installed between the ends of the two clamping rods (606) away from the limit plate (607), a clamping spring (609) is sleeved on the surface of the clamping rod (606) located between the movable frame (605) and the cleaning plate (608), and a limit slide rod (610) is slidably installed on both ends of the movable frame (605), and both ends of the limit slide rod (610) are fixedly connected to the inner wall of the liquid storage tank (402).

9. A method for using a circulating microfluidic device for cell sorting, according to any one of claims 1 to 8, characterized in that: The method of use comprises the following steps: According to the cell sorting requirements, a processing box (3) equipped with a suitable spiral flow channel (306) is selected, and the bottom of the processing box (3) is placed on the bottom of the installation base (2), so that the T-shaped card strip (301) at the bottom of the processing box (3) corresponds to the T-shaped card slot (201), and the processing box (3) is pushed to move on the top of the installation base (2) until the T-shaped card strip (301) is inserted into the corresponding T-shaped card slot (201), so that the processing box (3) gradually sticks to the concave base (1) and the side of the liquid storage box (4), until the top of the spiral flow channel (306) is tightly placed inside the drainage groove (404), and at the same time, the diversion rack (308) is inserted into the rectangular through groove (103), and the collection box (101) is inserted into the opening of the concave base (1), so that the rubber strip (102) is inserted into the collection box (1 01) are squeezed and deformed on both sides, thereby clamping and fixing the collection box (101) inside the concave base (1). When the processing box (3) gradually comes close to one side of the liquid storage tank (4), the fixed clamping block (304) and the fixed clamping plate (502) are tightly pressed together, and the fixed clamping plate (502) is lifted up to rotate and tilt inside the concave mounting frame (501), and the reset spring (505) is squeezed and deformed between the corresponding positioning protrusions (503) until the fixed clamping block (304) slides through the clamping plate (502). Under the rebound action of the reset spring (505), the fixed clamping plate (502) is lifted up and reset, so that the clamping plate (502) is clamped with the fixed clamping block (304), thereby clamping and fixing the processing box (3) on one side of the liquid storage tank (4); Flip open the second magnetic cover (401) and align the positioning slots (701) on both sides of the filter screen (7) with the positioning strips (403) on the inner wall of the liquid storage tank (402). Push the filter screen (7) into the liquid storage tank (402) so that the positioning slots (701) slide on the surface of the corresponding positioning strips (403) to ensure that the filter screen (7) and the cleaning plate (608) are in close contact. The filter screen (7) lifts up the cleaning plate (608) to squeeze and press the cleaning plate (608). The spring (609) is squeezed and deformed between the movable frame (605) and the cleaning plate (608), and the cleaning plate (608) is pressed against the surface of the filter screen plate (7) under the rebound action of the compression spring (609). An appropriate amount of cell stock solution is poured into the liquid storage tank (402) on one side of the filter screen plate (7), so that the cell stock solution is filtered through the filter screen plate (7) and flows to the conduit at the input end of the circulation pump (405) located inside the liquid storage tank (402). The magnetic cover plate (401) is closed. At the top of the liquid storage tank (4), the circulation pump (405) is started to pump the cell stock solution in the liquid storage tank (402) into the liquid drainage tank (404) through the conduit. The cell stock solution then flows from the liquid drainage tank (404) to the top of the spiral flow channel (306). When the cell stock solution flows in the spiral flow channel (306), cells of different sizes are sorted and blocked by the sorting bar (307) when the cell stock solution flows, so that cells of different sizes flow to both sides of the sorting bar (307) for sorting until they pass through the cell storage tank (402). The over-sorted cell stock solution and cells flow from the spiral flow channel (306) to the diversion rack (308), and are sorted by the diversion rack (308), the drainage rack 1 (309), and the drainage rack 2 (310), so that cells of different sizes are discharged from the drainage rack 1 (309) and the drainage rack 2 (310), and the excess cell stock solution flows from the diversion rack (308) to the collection box (101) inside the concave base (1) for centralized processing until the sorting of the cell stock solution inside the storage tank (402) is completed; At the same time, the filter plate (7) filters the cell debris in the cell stock solution, so that the cell debris accumulates on one side of the filter plate (7), and the rotating motor (604) is started to control the reciprocating screw (601) to rotate synchronously under the meshing action of the bevel gear 1 (602) and the bevel gear 2 (603), so that the movable frame (605) moves back and forth on the surface of the reciprocating screw (601), and at the same time, the two ends of the movable frame (605) slide on the surface of the corresponding limit slide rod (610), ensuring that the movable frame (605) does not tilt or deflect during the movement. The cleaning plate (608) moves with the movable frame (605) on one side of the filter plate (7) to clean the cell debris accumulated on the surface of the filter plate (7), so as to prevent the cell debris from accumulating on the surface of the filter plate (7) and affecting the filtering effect of the filter plate (7) on the cell stock solution.

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