A circulating microfluidic device for cell sorting and methods of use thereof

Abstract

This invention discloses a circulating microfluidic device for cell sorting and its usage method, belonging to the field of cell sorting technology. The device comprises a storage tank, a filter screen, a reciprocating screw, a moving frame, a cleaning plate, and a compression spring. After the positioning slots on both sides of the filter screen are engaged with the positioning strips on the inner wall of the storage tank, the filter screen and cleaning plate are pressed together, lifting the cleaning plate. This causes the compression spring to deform between the moving frame and the cleaning plate, and under the rebound of the compression spring, the cleaning plate is pressed firmly against the surface of the filter screen. An appropriate amount of cell stock solution is poured into the opening of the storage tank. Simultaneously, a rotating motor is started to control the reciprocating screw to rotate, causing the moving frame to reciprocate on the surface of the reciprocating screw, which in turn moves the cleaning plate back and forth on the surface of the filter screen. This cleans the cell debris accumulated on the surface of the filter screen, preventing cell debris from accumulating on the filter screen for a long time and clogging it, thus affecting the filtration effect of the cell stock solution.

Description

Technical Field

[0001] This invention relates to the field of cell sorting technology, specifically to a circulating microfluidic device for cell sorting and its method of use. Background Technology

[0002] Cell sorting plays a crucial role in numerous fields, including biology, medical research, and clinical diagnosis and treatment. Accurately and efficiently separating specific cell types from complex cell mixtures is of irreplaceable significance for gaining a deeper understanding of cell function, disease mechanisms, and developing targeted treatments. Microfluidic devices, as important equipment for cell sorting, achieve precise manipulation and sorting of cells through the design of specific microchannel structures and fluid drive methods. The cell stock solution flows in the channel, and the cells in the cell stock solution are separated into cells or particles of different sizes under the inertial force of the flowing cell stock solution, thereby sorting the cells in the cell stock solution according to cell size.

[0003] Existing circulating microfluidic devices for cell sorting suffer from several drawbacks when sorting cell stock solutions. The stock solutions contain a large amount of cell debris, which easily accumulates and clogs the filter screen during the initial filtration, reducing the filter's effectiveness and impacting the cell sorting process. Furthermore, since cell sorting devices are typically integrated units, it's difficult to easily change different flow channels to suit varying cell sorting requirements. Therefore, improvements are needed.

[0004] Based on this, the present invention designs a circulating microfluidic device for cell sorting and its usage method to solve the above problems. Summary of the Invention

[0005] The purpose of this invention is to provide a circulating microfluidic device for cell sorting and its usage method, in order to solve the problems mentioned in the background art. In the case of cell sorting, existing circulating microfluidic devices for cell sorting contain a large number of cell fragments. During the initial filtration, the cell fragments easily accumulate and clog the filter screen, which reduces the filtration effect of the filter screen on the cell stock solution and affects the cell sorting effect. At the same time, since the cell sorting device is generally a whole, it is difficult to change different flow channels at will according to different cell sorting requirements.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] A circulating microfluidic device for cell sorting includes a concave base. A mounting plate is fixedly installed on the bottom of the concave base on the side away from the opening. The top of the mounting plate has symmetrically arranged T-shaped slots. A processing box is provided on the top of the mounting plate. T-shaped clips are symmetrically fixedly installed on the bottom of the processing box, and the T-shaped clips are slidably connected to the inside of the T-shaped slots. A liquid storage tank is fixedly installed on the top of the concave base. A snap-fit ​​mechanism is fixedly installed on the top of both sides of the liquid storage tank near the processing box. A cleaning mechanism is provided inside the liquid storage tank. A filter screen is slidably installed inside the liquid storage tank on one side of the cleaning mechanism.

[0008] As a further embodiment of the present invention, a collection box is slidably installed through the opening on one side of the concave base, and rubber strips are symmetrically fixedly installed on the surfaces on both sides of the collection box. A rectangular through groove is provided on the side of the concave base away from the opening.

[0009] As a further embodiment of the present invention, the bottom of the arc surface of the processing box is provided with a drain port, and a block is inserted inside the drain port. Fixed blocks are fixedly installed on the top of the two side surfaces of the processing box respectively. A magnetic cover plate is rotatably installed on one side of the top of the processing box via a pin shaft. A spiral flow channel is fixedly installed on the inner wall of the processing box, and multiple sorting baffles are fixedly installed at equal intervals at the bottom of the track of the spiral flow channel.

[0010] As a further embodiment of the present invention, a diversion frame is fixedly installed at the bottom end of the spiral flow channel. The end of the diversion frame away from the spiral flow channel passes through the treatment box and corresponds to the position of the rectangular through groove. A first drain rack and a second drain rack are fixedly installed on both sides of the diversion frame, and the first drain rack and the second drain rack are respectively connected to both sides of the diversion frame. The ends of the first drain rack and the second drain rack away from the diversion frame pass through the treatment box and are fixedly connected to the treatment box.

[0011] As a further embodiment of the present invention, a magnetic cover plate is rotatably installed on one side of the top of the liquid storage tank via a pin. The liquid storage tank is provided with a liquid storage tank. Positioning strips are symmetrically fixedly installed on the inner wall of one side of the opening of the liquid storage tank. Positioning slots are provided on both sides of the filter screen, and the positioning slots are slidably connected to the surface of the positioning strips. A drain trough is provided on the side of the top of the liquid storage tank away from the opening of the liquid storage tank, and the position of the drain trough corresponds to the opening at the top of the spiral flow channel. A circulation pump is embedded and fixedly installed on the top of the liquid storage tank on the side of the drain trough. The input end and output end of the circulation pump are respectively connected to the conduit, and the conduit is respectively connected to the liquid storage tank and the drain trough.

[0012] As a further embodiment of the present invention, the snap-fit ​​mechanism includes a concave mounting bracket. A fixed snap plate is rotatably mounted on one side of the concave mounting bracket via a pin, and the snap-fit ​​opening of the fixed snap plate corresponds to the position of the fixed snap block. Positioning protrusions are symmetrically fixedly mounted on the sides of the concave mounting bracket and the fixed snap plate away from the snap-fit ​​opening of the fixed snap plate, and the positioning protrusions on the surfaces of the concave mounting bracket and the fixed snap plate are opposite each other. Positioning sleeves are fixedly mounted on the surfaces of the concave mounting bracket and the fixed snap plate located on the side of the positioning protrusions, and a return spring is sleeved between the two opposite positioning protrusions, and the two ends of the return spring are snapped into the inside of the positioning sleeve.

[0013] As a further embodiment of the present invention, the cleaning mechanism includes a reciprocating lead screw, the two ends of which are rotatably connected to the inner wall of the liquid storage tank via bearings. One end of the reciprocating lead screw passes through the liquid storage tank and is fixedly installed with a bevel gear one. A bevel gear two meshes with the surface of the bevel gear one. A rotating motor is fixedly installed on one side of the bevel gear two and is fixedly connected to the outer surface of the liquid storage tank. A movable frame is threaded through the surface of the reciprocating lead screw.

[0014] As a further embodiment of the present invention, a clamping rod is slidably installed through both sides of the movable frame. One end of the clamping rod passes through the movable frame and is fixedly installed with a limiting plate. A cleaning plate is fixedly installed between the ends of the two clamping rods away from the limiting plate. A clamping spring is sleeved on the surface of the clamping rod located between the movable frame and the cleaning plate. Limiting slide rods are slidably installed through both ends of the movable frame, and the two ends of the limiting slide rods are fixedly connected to the inner wall of the liquid storage tank.

[0015] A method of using a circulating microfluidic device for cell sorting, the method comprising the following steps:

[0016] Select a processing box equipped with a suitable spiral flow channel according to the cell sorting requirements. Place the bottom of the processing box against the bottom of the mounting base, aligning the T-shaped retaining strips on the bottom of the processing box with the T-shaped retaining slots. Push the processing box on top of the mounting base until the T-shaped retaining strips are inserted into the corresponding T-shaped retaining slots, gradually bringing the processing box closer to the concave base and one side of the storage tank, until the top of the spiral flow channel is firmly against the inside of the drainage trough. Simultaneously, insert the diverter rack into the rectangular through-slot and insert the collection box into the opening of the concave base, ensuring the rubber strips are on both sides of the collection box. The compression deformation clamps the collection box tightly inside the concave base. As the treatment box gradually presses against one side of the storage tank, the fixing block and the fixing plate latch are pressed together, lifting the fixing plate to rotate and tilt inside the concave mounting bracket. The compression return spring is compressed and deformed between the corresponding positioning protrusions until the fixing block slides past the latch of the fixing plate. Under the rebound action of the return spring, the fixing plate is lifted and reset, so that the latch of the fixing plate engages with the fixing block, thereby clamping and fixing the treatment box to one side of the storage tank.

[0017] Flip open the magnetic cover plate two and align the positioning slots on both sides of the filter plate with the positioning strips on the inner wall of the storage tank. Push the filter plate into the storage tank, allowing the positioning slots to slide against the corresponding positioning strips, ensuring the filter plate is firmly attached to the cleaning plate. The filter plate lifts the cleaning plate, causing the cleaning plate to compress the clamping spring between the moving frame and the cleaning plate. Under the rebound of the clamping spring, the cleaning plate is pressed firmly against the surface of the filter plate. Pour an appropriate amount of cell stock solution into the storage tank on the side of the filter plate, allowing the cell stock solution to flow through the filter plate to the conduit at the input end of the circulation pump located inside the storage tank. Close the magnetic cover plate two on top of the storage tank and start the circulation pump. The circular pump pumps the cell stock solution from the storage tank into the drainage tank through a conduit. The cell stock solution then flows from the drainage tank to the top of the spiral channel. As the cell stock solution flows inside the spiral channel, cells of different sizes are separated and blocked by the sorting baffles, causing cells of different sizes to flow to both sides of the sorting baffles for further separation. The separated cell stock solution and cells flow from the spiral channel to the distribution rack. After being separated by the distribution rack, drainage rack one, and drainage rack two, cells of different sizes are discharged from drainage rack one and drainage rack two. Excess cell stock solution flows from the distribution rack to the collection box inside the concave base for centralized processing until the cell stock solution in the storage tank is completely separated.

[0018] Simultaneously, the filter plate filters cell debris from the cell stock solution, causing the cell debris to accumulate on one side of the filter plate. The rotating motor is started, and the reciprocating screw is controlled to rotate synchronously under the meshing action of bevel gear one and bevel gear two, causing the moving frame to move back and forth on the surface of the reciprocating screw. At the same time, the two ends of the moving frame slide on the corresponding limit slide bar surfaces to ensure that the moving frame does not tilt or deflect during the movement. The cleaning plate moves with the moving frame on one side of the filter plate to clean the cell debris accumulated on the surface of the filter plate, preventing cell debris from accumulating on the surface of the filter plate and affecting the filtration effect of the filter plate on the cell stock solution.

[0019] Compared with the prior art, the beneficial effects of the present invention are:

[0020] 1. This invention comprises a storage tank, a filter screen, a reciprocating screw, a moving frame, a cleaning plate, and a compression spring. After the filter screen is inserted into the storage tank, with the positioning slots on both sides of the filter screen corresponding to the positioning strips on the inner wall of the storage tank, the filter screen and cleaning plate are pressed together, lifting the cleaning plate. This causes the compression spring to deform between the moving frame and the cleaning plate. Under the rebound of the compression spring, the cleaning plate is pressed firmly against the surface of the filter screen. An appropriate amount of cell stock solution is poured into the opening of the storage tank. Simultaneously, a rotating motor is activated to control the reciprocating screw to rotate, causing the moving frame to reciprocate on the surface of the reciprocating screw. This drives the cleaning plate to move back and forth on the surface of the filter screen, cleaning the cell debris accumulated on the surface of the filter screen. This prevents cell debris from accumulating on the filter screen for a long time and clogging it, thus affecting the filtration effect of the cell stock solution.

[0021] 2. This invention, by setting up an installation base plate, a processing box, fixing blocks, a fixing plate, and a return spring, allows the processing box to be placed on top of the installation base plate, aligning the T-shaped locking strips at the bottom of the processing box with the T-shaped locking slots. This pushes the processing box to move on top of the installation base plate until the T-shaped locking strips are inserted into the corresponding T-shaped locking slots, causing the processing box to gradually approach the storage tank. The fixing blocks on both sides of the processing box then approach the fixing plate, lifting the fixing plate and causing it to rotate. This causes the return spring to deform between the corresponding positioning protrusions until the fixing blocks pass the locking slots of the fixing plate. The return spring then rebounds, lifting the fixing plate so that the locking slots of the fixing plate engage with the fixing blocks, thus securing the processing box tightly to one side of the storage tank and fixing it to the top of the installation base plate. This allows for easy replacement of the appropriate processing box according to cell sorting needs.

[0022] 3. This invention utilizes a circulating pump, a drain tank, a spiral flow channel, sorting baffles, a diverter, and two drain racks. The circulating pump pumps the cell stock solution, filtered through a filter screen, from the storage tank into the drain tank. The cell stock solution gradually flows towards the top of the spiral flow channel, causing it to flow within the channel. Due to the different sizes of cells in the cell stock solution, the inertial forces acting on the flowing cells cause cells of different sizes to flow towards the sorting baffles for sorting. After sorting, the cells and cell stock solution flow along the spiral flow channel to the diverter. Excess cell stock solution flows from the diverter into a collection box for centralized processing. Cells of different sizes are then discharged from the processing tank through drain racks one and two, thus achieving the sorting of cells of different sizes. Attached Figure Description

[0023] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0024] Figure 1 This is a schematic diagram of the unfolded structure of the present invention;

[0025] Figure 2 This is a schematic diagram of the unfolded structure of the concave base and mounting plate of the present invention;

[0026] Figure 3 This is a cross-sectional view of the processing box and spiral flow channel of the present invention.

[0027] Figure 4 This is a schematic diagram of the spiral flow channel and sorting baffle of the present invention;

[0028] Figure 5This is a schematic diagram of the unfolded structure of the liquid storage tank and cleaning mechanism of the present invention;

[0029] Figure 6 This is a schematic diagram of the snap-fit ​​mechanism of the present invention;

[0030] Figure 7 This is a cross-sectional view of the reciprocating lead screw and the moving frame of the present invention;

[0031] Figure 8 This is a schematic diagram of the structure of the filter screen of the present invention.

[0032] The attached diagram lists the components represented by each number as follows:

[0033] 1. Concave base; 101. Collection box; 102. Rubber strip; 103. Rectangular through groove; 2. Mounting base plate; 201. T-shaped slot; 3. Processing box; 301. T-shaped clip; 302. Drain outlet; 303. Block; 304. Fixing clip; 305. Magnetic cover plate one; 306. Spiral flow channel; 307. Sorting baffle; 308. Diverter; 309. Drain rack one; 310. Drain rack two; 4. Storage tank; 401. Magnetic cover plate two; 402. Storage tank; 403. Positioning clip; 4 04. Drainage tank; 405. Circulation pump; 5. Snap-fit ​​mechanism; 501. Concave mounting bracket; 502. Fixing plate; 503. Positioning protrusion; 504. Positioning sleeve; 505. Return spring; 6. Cleaning mechanism; 601. Reciprocating screw; 602. Bevel gear one; 603. Bevel gear two; 604. Rotary motor; 605. Moving frame; 606. Pressing rod; 607. Limiting plate; 608. Cleaning plate; 609. Pressing spring; 610. Limiting slide bar; 7. Filter screen; 701. Positioning slot. Detailed Implementation

[0034] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0035] Please see Figures 1-8 The present invention provides a technical solution:

[0036] A circulating microfluidic device for cell sorting includes a concave base 1, a mounting base plate 2 fixedly installed on the bottom of the concave base 1 away from the opening, a T-shaped slot 201 extending to the edge symmetrically provided on the top of the mounting base plate 2, a processing box 3 provided on the top of the mounting base plate 2, a T-shaped clip 301 symmetrically fixedly installed on the bottom of the processing box 3, and the T-shaped clip 301 slidingly connected to the inside of the T-shaped slot 201; a liquid storage tank 4 fixedly installed on the top of the concave base 1, and a snap-fit ​​mechanism 5 fixedly installed on the top of both sides of the liquid storage tank 4 near the processing box 3; a cleaning mechanism 6 provided inside the liquid storage tank 4, and a filter screen 7 slidingly installed inside the liquid storage tank 4 on one side of the cleaning mechanism 6;

[0037] During operation, select the appropriate processing box 3 according to the cell sorting requirements, place the processing box 3 on top of the mounting base plate 2 and push it so that the T-shaped clip 301 at the bottom of the processing box 3 is inserted into the corresponding T-shaped slot 201. Ensure that the processing box 3 will not be misaligned when it is fixed on the top of the mounting base plate 2 until the processing box 3 is close to one side of the liquid storage tank 4. Then, the snap-fit ​​mechanism 5 will snap the processing box 3 in place and fix it on the top of the mounting base plate 2. Insert the filter plate 7 into the storage tank 4, ensuring it is tightly attached to the cleaning mechanism 6. Pour an appropriate amount of cell stock solution into the storage tank 4. The cell stock solution, after being filtered by the filter plate 7, flows to one side of the storage tank 4 and is pressurized and pumped into the top of the processing tank 3 for sorting. The sorted cell stock solution is discharged from the processing tank 3. Some of the excess cell stock solution flows into the concave base 1 for centralized processing. At the same time, the cleaning mechanism 6 is activated to clean the cell debris accumulated on the surface of the filter plate 7, preventing cell debris from accumulating on the surface of the filter plate 7 and affecting the filtration of the cell stock solution by the filter plate 7.

[0038] As a further embodiment 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 on 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.

[0039] During operation, the collection box 101 is inserted into the opening of the concave base 1, causing the rubber strips 102 on both sides of the collection box 101 to be squeezed and deformed against the inner wall of the concave base 1, thereby clamping and fixing the collection box 101 inside the concave base 1. This collects and concentrates the cell stock solution dispensed from the rectangular through groove 103, preventing leakage of excess cell stock solution during discharge. At the same time, the concentrated cell stock solution can be subjected to secondary sorting, improving the cell sorting effect.

[0040] As a further embodiment of the present invention, a drain port 302 is provided at the bottom of the arc surface of the processing box 3, and a block 303 is snapped into the drain port 302. A magnetic cover plate 305 is rotatably installed on one side of the top of the processing box 3 via a pin. A spiral flow channel 306 is fixedly installed on the inner wall of the processing box 3. Multiple sorting baffles 307 are fixedly installed at equal intervals at the bottom of the track of the spiral flow channel 306. A diverter 308 is fixedly installed at the bottom end of the spiral flow channel 306. The end of the diverter 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. Drainage rack 1 309 and drainage rack 2 310 are fixedly installed on both sides of the diversion rack 308, and drainage rack 1 309 and drainage rack 2 310 are respectively connected to both sides of the diversion rack 308. In this embodiment, drainage rack 1 309, drainage rack 2 310 and diversion rack 308 are U-shaped sections. The ends of drainage rack 1 309 and drainage rack 2 310 away from the diversion rack 308 respectively penetrate through the processing box 3 and are fixedly connected to the processing box 3.

[0041] During operation, the cell culture medium is introduced from the top of the spiral channel 306 and flows inside the spiral channel 306. Due to the different cell sizes, cells of different sizes are guided by the flow of the cell culture medium to both sides of the sorting baffles 307 for sorting. After sorting, the cells and cell culture medium flow to the diversion rack 308. The sorted cells are discharged from the discharge rack 1 309 and the discharge rack 2 310 into the processing tank 3 for collection. Excess cell culture medium flows from the diversion rack 308 into the collection box 101 for centralized processing. At the same time, the cell culture medium is prone to splashing out and falling into the processing tank 3 when flowing inside the spiral channel 306. After sorting is completed, the processing tank 3 is removed from the top of the mounting base plate 2. The block is removed, and the block 303 is simultaneously extracted from the drain port 302, thereby pouring out the splashed cell concentrate inside the processing tank 3 to prevent it from deteriorating due to prolonged exposure. The cell concentrate flows inside the spiral channel 306. During this flow, because the sorting baffle 307 is fixed inside the spiral channel 306, and because the inner ring of the spiral channel 306 is lower than the outer ring, the end of the sorting baffle 307 near the outer side of the spiral channel 306 is higher. When cell fragments in the cell concentrate are blocked and screened by the sorting baffle 307, the cell concentrate impacts the sorting baffle 307 and is pushed towards the sorting baffle 307. The flow impact force generated on both sides of 07 causes smaller cell fragments to flow with the flowing cell concentrate towards the higher end of the sorting baffle 307, while larger cell fragments flow towards the lower end. When the cell concentrate flows within the spiral channel 306 and impacts the sorting baffle 307, the spiral structure of the channel causes centrifugal force. Larger cell fragments experience greater centrifugal force and are thrown towards the outer side of the spiral channel 306, while smaller cell fragments experience less centrifugal force and are distributed in the middle or inner side of the spiral channel 306, where they are blocked by the sorting baffle 307. As the cell concentrate flows... The flow of the liquid causes smaller cell fragments to flow along with the original cell solution to the inner side of the spiral channel 306. This allows the original solution flowing out of the spiral channel 306 to be discharged from the diversion rack 308 via the first drain rack 309 and the second drain rack 310. The diversion rack 308 has two symmetrical diversion channels inside, which are connected to the first drain rack 309 and the second drain rack 310, respectively. This allows the original solution flowing out of the spiral channel 306 to be diverted, so that the original solution containing larger cell fragments flows to the first drain rack 309, and the original solution containing smaller cell fragments flows to the second drain rack 310. Excess original solution is discharged from the opening of the diversion rack 308.

[0042] As a further embodiment of the present invention, a magnetic cover plate 401 is rotatably mounted on one side of the top of the liquid storage tank 4 via a pin. The liquid storage tank 4 has a liquid storage tank 402 inside. Positioning strips 403 are symmetrically fixedly mounted on the inner wall of the top opening of the liquid storage tank 402. Positioning slots 701 are respectively provided on both sides of the filter screen 7, and the positioning slots 701 are slidably connected to the surface of the positioning strips 403. The filter screen 7 divides the liquid storage tank 402 into two spaces. A drain trough 404 is provided on the top side of the liquid storage tank 4 away from the opening of the liquid storage tank 402, and the drain trough 404 corresponds to the opening at the top of the spiral flow channel 306. A circulation pump 405 is embedded and fixedly mounted on the top of the liquid storage tank 4 on one side of the drain trough 404. The input and output ends of the circulation pump 405 are respectively connected to conduits, and the conduits are respectively connected to one space of the liquid storage tank 402 and the drain trough 404.

[0043] During operation, flip the magnetic cover plate 401 to open the top of the storage tank 4, then insert the filter plate 7 into the storage tank 402, so that the positioning slots 701 on both sides of the filter plate 7 are locked onto the surface of the positioning strip 403, thereby filtering the cell stock solution poured into the storage tank 402. The filtered cell stock solution flows to the other side of the storage tank 402. Start the circulation pump 405, so that the conduit at the input end of the circulation pump 405 pumps the filtered cell stock solution in the storage tank 402 into the drain tank 404, thereby guiding the filtered cell stock solution into the spiral flow channel 306 for sorting. At the same time, flip the magnetic cover plate 401 to stick it tightly to the top of the storage tank 4, so that the magnetic cover plate 401 sticks tightly to the top of the storage tank 4, preventing impurities in the air on the surface from falling into the storage tank 402 and causing the cell stock solution to deteriorate.

[0044] As a further embodiment of the present invention, fixed blocks 304 are fixedly installed on the top of both sides of the processing box 3; the snap-fit ​​mechanism 5 includes a concave mounting bracket 501, a fixed plate 502 is rotatably installed on one side of the concave mounting bracket 501 via a pin, and the snap-fit ​​at one end of the fixed plate 502 corresponds to the position of the fixed block 304; positioning protrusions 503 are symmetrically fixedly installed on the side of the concave mounting bracket 501 and the fixed plate 502 away from the snap-fit ​​of the fixed plate 502, and the positioning protrusions 503 on the surface of the concave mounting bracket 501 and the fixed plate 502 are opposite to each other; positioning sleeves 504 are fixedly installed on the surface of the concave mounting bracket 501 and the fixed plate 502 on the side of the positioning protrusions 503, and a return spring 505 is sleeved between the two opposite positioning protrusions 503, and the two ends of the return spring 505 are snapped into the inside of the positioning sleeve 504;

[0045] During operation, when the processing box 3 is pushed close to the storage tank 4, the fixing block 304 gradually presses against the fixing plate 502, lifting one end of the fixing plate 502. This causes the return spring 505 to be squeezed and deformed between the corresponding positioning protrusions 503. At the same time, the two ends of the return spring 505 move inside the corresponding positioning sleeve 504, ensuring that the two ends of the return spring 505 will not be misaligned or shifted, causing the return spring 505 to detach from the surface of the positioning protrusion 503. This continues until the fixing block 304 slides past the slot of the fixing plate 502, at which point the return spring 505 rebounds and lifts the fixing plate 502 to rotate and reset, thus clamping the fixing plate 502 onto the surface of the fixing block 304, thereby clamping and fixing the processing box 3 to one side of the storage tank 4.

[0046] As a further embodiment of the present invention, the cleaning mechanism 6 includes a reciprocating lead screw 601. Both ends of the reciprocating lead screw 601 are rotatably connected to the inner wall of the liquid storage tank 402 via bearings. One end of the reciprocating lead screw 601 passes through the liquid storage tank 4 and is fixedly mounted with a first bevel gear 602. A second bevel gear 603 meshes with the surface of the first bevel gear 602. A rotating motor 604 is fixedly mounted on one side of the second bevel gear 603 and is fixedly connected to the outer surface of the liquid storage tank 4. A movable frame 605 is threadedly connected through the surface of the reciprocating lead screw 601. A clamping rod 606 is slidably installed through 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 limiting plate 607. A cleaning plate 608 is fixedly installed between the ends of the two clamping rods 606 away from the limiting plate 607. A clamping spring 609 is sleeved on the surface of the clamping rod 606 between the movable frame 605 and the cleaning plate 608. A limiting slide rod 610 is slidably installed through both ends of the movable frame 605, and both ends of the limiting slide rod 610 are fixedly connected to the inner wall of the liquid storage tank 402.

[0047] During operation, after inserting the filter screen plate 7 into the liquid storage tank 402 and ensuring it is pressed against the cleaning plate 608, the filter screen plate 7 lifts up the cleaning plate 608, causing the compression spring 609 to deform between the moving frame 605 and the cleaning plate 608. Under the rebound action of the compression spring 609, the cleaning plate 608 is pressed firmly onto the surface of the filter screen plate 7. The rotating motor 604 is started to control the second bevel gear 603 to rotate. Under the meshing action of the second bevel gear 603 and the first bevel gear 602, the reciprocating screw 601 is controlled to rotate synchronously, causing the moving frame 605 to move back and forth on the surface of the limit slide bar 610 with the rotation of the reciprocating screw 601. This drives the cleaning plate 608 to move on the surface of the filter screen plate 7, pushing the cell debris accumulated on one side of the filter screen plate 7 to move, thus preventing cell debris from accumulating on the surface of the filter screen plate 7 and affecting the filtration effect of the filter screen plate 7.

[0048] A method of using a circulating microfluidic device for cell sorting, comprising the following steps:

[0049] Select a processing box 3 equipped with a suitable spiral flow channel 306 according to the cell sorting requirements. Place the bottom of the processing box 3 on the bottom of the mounting base 2, aligning the T-shaped retaining strip 301 on the bottom of the processing box 3 with the T-shaped retaining groove 201. Push the processing box 3 on the top of the mounting base 2 until the T-shaped retaining strip 301 is inserted into the corresponding T-shaped retaining groove 201, gradually pressing the processing box 3 against the concave base 1 and the side of the storage tank 4, until the top of the spiral flow channel 306 is pressed against the inside of the drain trough 404. At the same time, the diverter 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. As the processing box 3 gradually presses against one side of the storage tank 4, the fixing block 304 is pressed against the locking slot of the fixing plate 502, and the fixing plate 502 is lifted up and rotated inside the concave mounting bracket 501. The compression return spring 505 is squeezed and deformed between the corresponding positioning protrusions 503 until the fixing block 304 slides past the locking slot of the fixing plate 502. Under the rebound action of the return spring 505, the fixing plate 502 is lifted and reset, so that the locking slot of the fixing plate 502 is engaged with the fixing block 304, thereby clamping and fixing the processing box 3 to one side of the storage tank 4.

[0050] Flip open the magnetic cover 401 and align the positioning slots 701 on both sides of the filter plate 7 with the positioning strips 403 on the inner wall of the storage tank 402. Push the filter plate 7 into the storage tank 402, so that the positioning slots 701 slide against the corresponding positioning strips 403, ensuring that the filter plate 7 is tightly attached to the cleaning plate 608. The filter plate 7 lifts the cleaning plate 608, causing the cleaning plate 608 to compress the clamping spring 609 between the moving frame 605 and the cleaning plate 608, deforming it. Under the rebound of the clamping spring 609, the cleaning plate 608 is pressed firmly onto the surface of the filter plate 7. Pour an appropriate amount of cell stock solution into the storage tank 402 on one side of the filter plate 7, so that the cell stock solution flows through the filter plate 7 to the conduit at the input end of the circulation pump 405 located inside the storage tank 402. Close the magnetic cover 401 on top of the storage tank 4. The circulation pump 405 is started to pump the cell stock solution inside the storage tank 402 into the drainage tank 404 through the conduit. The cell stock solution then flows from the drainage tank 404 to the top of the spiral channel 306. When the cell stock solution flows inside the spiral channel 306, cells of different sizes are separated and blocked by the sorting baffles 307, causing cells of different sizes to flow to both sides of the sorting baffles 307 for sorting, until the sorted cell stock solution and cells flow from the spiral channel 306 to the diversion rack 308. After being sorted by the diversion rack 308, the first drainage rack 309 and the second drainage rack 310, cells of different sizes are discharged from the first drainage rack 309 and the second drainage rack 310. 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 cell stock solution in the storage tank 402 is sorted.

[0051] Simultaneously, the filter plate 7 filters cell debris from the cell stock solution, causing the cell debris to accumulate on one side of the filter plate 7. The rotating motor 604 is started, and the reciprocating screw 601 is rotated synchronously under the meshing action of bevel gear 1 602 and bevel gear 2 603, causing the moving frame 605 to move back and forth on the surface of the reciprocating screw 601. At the same time, the two ends of the moving frame 605 slide on the surface of the corresponding limit slide rod 610 to ensure that the moving frame 605 does not tilt or deflect during the movement. The cleaning plate 608 moves with the moving 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 avoid the accumulation of cell debris on the surface of the filter plate 7 affecting the filtration effect of the filter plate 7 on the cell stock solution.

Claims

1. A circulating microfluidic device for cell sorting, comprising a concave base (1), characterized in that: The concave base (1) has a mounting base plate (2) fixedly installed on the bottom of the side away from the opening. The mounting base plate (2) has a T-shaped slot (201) symmetrically provided on the top of the mounting base plate (2). The mounting base plate (2) has a processing box (3) on the top. The processing box (3) has a T-shaped clip (301) symmetrically fixedly installed on the bottom of the processing box (3). The T-shaped clip (301) is slidably connected to the inside of the T-shaped slot (201). The concave base (1) has a liquid storage tank (4) fixedly installed on the top of the concave base (1). The top of the two sides of the liquid storage tank (4) near the processing box (3) is fixedly installed with a snap-fit ​​mechanism (5). The liquid storage tank (4) has a cleaning mechanism (6) inside. The liquid storage tank (4) has a filter screen plate (7) slidably installed inside the liquid storage tank (4) on one side of the cleaning mechanism (6). The bottom of the arc surface of the processing box (3) is provided with a drain port (302), and a block (303) is snapped into the drain port (302). Fixed blocks (304) are fixedly installed on the top of the two sides of the processing box (3). 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). Multiple sorting baffles (307) are fixedly installed at equal intervals in the bottom of the track of the spiral flow channel (306). A diversion frame (308) is fixedly installed at the bottom end of the spiral flow channel (306). The end of the diversion frame (308) away from the spiral flow channel (306) passes through the treatment box (3) and corresponds to the position of the rectangular through groove (103). Drainage rack one (309) and drainage rack two (310) are fixedly installed on both sides of the diversion frame (308), and drainage rack one (309) and drainage rack two (310) are respectively connected to both sides of the diversion frame (308). The ends of drainage rack one (309) and drainage rack two (310) away from the diversion frame (308) pass through the treatment box (3) and are fixedly connected to the treatment box (3). The snap-fit ​​mechanism (5) includes a concave mounting bracket (501). A fixed clamping plate (502) is rotatably mounted on one side of the concave mounting bracket (501) via a pin shaft. The snap-fit ​​opening of the fixed clamping plate (502) corresponds to the position of the fixed clamping block (304). Positioning protrusions (503) are symmetrically fixedly mounted on the side of the concave mounting bracket (501) and the fixed clamping plate (502) away from the snap-fit ​​opening of the fixed clamping plate (502). The positioning protrusions (503) on the surfaces of the concave mounting bracket (501) and the fixed clamping plate (502) are opposite to each other. Positioning sleeves (504) are fixedly mounted on the surfaces of the concave mounting bracket (501) and the fixed clamping plate (502) on the side of the positioning protrusions (503). A return spring (505) is sleeved between the two opposite positioning protrusions (503), and the two ends of the return spring (505) are snapped into the inside of the positioning sleeve (504). The cleaning mechanism (6) includes a reciprocating screw (601), the two ends of which are rotatably connected to the inner wall of the liquid storage tank (402) via bearings. One end of the reciprocating screw (601) passes through the liquid storage tank (4) and is fixedly installed with a bevel gear (602). A bevel gear (603) meshes with the surface of the bevel gear (602). A rotating motor (604) is fixedly installed on one side of the bevel gear (603) and is fixedly connected to the outer surface of the liquid storage tank (4). A movable frame (605) is threaded through the surface of the reciprocating screw (601). A clamping rod (606) is slidably installed through 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 limiting plate (607). A cleaning plate (608) is fixedly installed between the ends of the two clamping rods (606) away from the limiting plate (607). A clamping spring (609) is sleeved on the surface of the clamping rod (606) between the movable frame (605) and the cleaning plate (608). A limiting slide rod (610) is slidably installed through both ends of the movable frame (605), and both ends of the limiting slide rod (610) are fixedly connected to the inner wall of the liquid storage tank (402).

2. The circulating microfluidic device for cell sorting according to claim 1, characterized in that: A collection box (101) is slidably installed through the opening on one side of the concave base (1). Rubber strips (102) are symmetrically fixed on the surfaces on 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.

3. A circulating microfluidic device for cell sorting according to claim 1, characterized in that: A magnetic cover plate (401) is rotatably installed on one side of the top of the liquid storage tank (4) via a pin. The liquid storage tank (4) is provided with a liquid storage tank (402) inside. A positioning strip (403) is symmetrically fixedly installed on the inner wall of one side of the opening of the liquid storage tank (402). A positioning slot (701) is provided on both sides of the filter screen plate (7), and the positioning slot (701) is slidably connected to the surface of the positioning strip (403). A drain trough (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 position of the drain trough (404) corresponds to the opening at the top of the spiral flow channel (306). A circulation pump (405) is embedded and fixedly installed on the top of the liquid storage tank (4) on one side of the drain trough (404). The input end and output end of the circulation pump (405) are respectively connected to the conduit, and the conduit is respectively connected to the liquid storage tank (402) and the drain trough (404).

4. A method of using a circulating microfluidic device for cell sorting, comprising the circulating microfluidic device for cell sorting as described in any one of claims 1-3, characterized in that, The method of use includes the following steps: According to the cell sorting requirements, select a processing box (3) equipped with a suitable spiral flow channel (306), place the bottom of the processing box (3) on the bottom of the mounting base plate (2), so that the T-shaped clip (301) at the bottom of the processing box (3) corresponds to the T-shaped slot (201), push the processing box (3) to move on the top of the mounting base plate (2) until the T-shaped clip (301) is inserted into the corresponding T-shaped slot (201), so that the processing box (3) gradually adheres to the concave base (1) and the side of the liquid storage tank (4), until the top of the spiral flow channel (306) adheres to the inside of the drain trough (404), at the same time the diverter (308) is inserted into the rectangular through groove (103), insert the collection box (101) into the opening of the concave base (1), so that the rubber strip (102) is in the collection box (101). 01) The two sides are squeezed and deformed, thereby clamping and fixing the collection box (101) inside the concave base (1). When the treatment box (3) gradually presses against the side of the storage tank (4), the fixing block (304) and the fixing plate (502) are pressed together, and the fixing plate (502) is lifted up and rotated inside the concave mounting bracket (501). The compression reset spring (505) is squeezed and deformed between the corresponding positioning protrusions (503) until the fixing block (304) slides through the slot of the fixing plate (502). Under the rebound action of the reset spring (505), the fixing plate (502) is lifted up and reset, so that the slot of the fixing plate (502) is engaged with the fixing block (304), thereby clamping and fixing the treatment box (3) to the side of the storage tank (4). Flip open the magnetic cover plate 2 (401) and align the positioning slots (701) on both sides of the filter screen plate (7) with the positioning clips (403) on the inner wall of the liquid storage tank (402). Push the filter screen plate (7) into the liquid storage tank (402) so that the positioning slots (701) slide on the surface of the corresponding positioning clips (403) to ensure that the filter screen plate (7) is close to the cleaning plate (608). The filter screen plate (7) lifts up the cleaning plate (608) so that the cleaning plate (608) is squeezed and pressed tightly. The spring (609) is squeezed and deformed between the moving frame (605) and the cleaning plate (608). Under the rebound action of the compression spring (609), the cleaning plate (608) is pressed against the surface of the filter screen plate (7). An appropriate amount of cell stock solution is poured into the storage tank (402) on one side of the filter screen plate (7). The cell stock solution flows through the filter screen plate (7) to the conduit at the input end of the circulation pump (405) located inside the storage tank (402). The magnetic cover plate two (401) is closed. At the top of the storage tank (4), the circulation pump (405) is started to pump the cell stock solution inside the storage tank (402) into the drainage tank (404) through the conduit. The cell stock solution then flows from the drainage tank (404) to the top of the spiral channel (306). When the cell stock solution flows inside the spiral channel (306), cells of different sizes are separated and blocked by the sorting baffles (307), causing cells of different sizes to flow to both sides of the sorting baffles (307) for sorting, until they pass through the separation baffles (307). The over-sorted cell stock solution and cells flow from the spiral flow channel (306) to the diversion rack (308). After being sorted by the diversion rack (308), the first drain rack (309) and the second drain rack (310), cells of different sizes are discharged from the first drain rack (309) and the second drain rack (310). 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 cell stock solution inside the storage tank (402) is sorted. Meanwhile, the filter plate (7) filters the cell debris in the cell stock solution, causing the cell debris to accumulate on one side of the filter plate (7). The rotating motor (604) is started, and the reciprocating screw (601) is controlled to rotate synchronously under the meshing action of bevel gear one (602) and bevel gear two (603), so that the moving frame (605) moves back and forth on the surface of the reciprocating screw (601). At the same time, the two ends of the moving frame (605) slide on the surface of the corresponding limit slide bar (610) to ensure that the moving frame (605) will not tilt or deflect during the movement. The cleaning plate (608) moves with the moving frame (605) on one side of the filter plate (7) to clean the cell debris accumulated on the surface of the filter plate (7) and prevent the cell debris from accumulating on the surface of the filter plate (7) and affecting the filtration effect of the filter plate (7) on the cell stock solution.

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