Suction filtration device for CTC enrichment dyeing

Abstract

The invention relates to the technical field of medical inspection equipment, in particular to a suction filtration device for CTC enrichment staining, which comprises a fixing assembly and a micro-fluidic chip mounted on the fixing assembly, the micro-fluidic chip comprises a reagent groove and a rigid chip fixing sheet arranged at the lower end of the reagent groove, the rigid chip fixing sheet is provided with a first through hole, and a second through hole is formed in the rigid chip fixing sheet. A chip body is placed in the first through hole, the reagent groove is of a sheet structure with a liquid leakage hole longitudinally penetrating through the body of the reagent groove, and a liquid suction assembly is arranged on the lower surface of the rigid chip fixing sheet. The suction filtration device adopts a rigid chip which is uniform in aperture and less in filtrate residue, so that the phenomenon of cell stacking or hardening caused by overlarge negative pressure during suction filtration can be effectively avoided, and the quality of cell enrichment and dyeing is improved; residual reagents in the chip can be removed by swinging and inclining the fixing assembly, so that automatic operation can be conveniently realized.

Description

Technical Field

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

[0002] Circulating tumor cell detection, referred to as CTC, is an emerging tumor diagnostic detection technology that has emerged in recent years. According to research, circulating tumor cells can be found in peripheral blood before solid tumors form, so CTC detection is very suitable for early screening and early diagnosis of malignant tumors, and CTC detection has a good effect on the prognosis of malignant tumors, disease progression monitoring, recurrence prediction, postoperative micro-lesion monitoring of malignant tumors, and the design and treatment effect monitoring of targeted drug therapy. It is currently an advanced method for early screening and diagnosis of malignant tumors. However, due to the low content of circulating tumor cells in peripheral blood, the capture and identification of circulating tumor cells in CTC detection is a great challenge. Microfluidic chips, also known as microporous membranes, are key components for enriching circulating tumor cells. Existing microporous membranes all use filter membranes or filter paper made of flexible fiber materials as filter materials. For example, another Chinese patent 202321916277X of the applicant discloses "A microfluidic device and cell enrichment and staining integrated device", and the authorization announcement number is CN220371064U, in which the microfluidic device includes: an effusion liner, a filter component, and a liner net. The interior of the effusion liner is a funnel-shaped structure, and a guide strip protruding from the surface of the funnel-shaped structure is provided for guiding the filtrate; the filter assembly is a microfiltration membrane provided above the effusion liner, and is used for filtering liquid samples. The pore size of the microfiltration membrane made of flexible materials is uneven, the capillary phenomenon is obvious, and wrinkles are easy to appear. The filtrate is easily retained in the filter membrane. Therefore, it is necessary to set auxiliary devices such as a negative pressure pump and a water absorption layer below to absorb the excess filtrate in the microfiltration membrane. If the negative pressure is too large, it is easy to cause cell stacking or hardening. The use of a rigid membrane can effectively avoid wrinkles, and no bubbles will appear during the enrichment and staining process, which will not affect the subsequent microscopic observation; but on the other hand, due to the lack of negative pressure suction filtration, this rigid microfluidic chip can only use swinging or tilting methods to remove the residual reagents in the chip, and the existing microfluidic chip fixing devices cannot achieve swinging and tilting movements. Summary of the invention

[0003] In order to solve the above-mentioned problems of the prior art, the present invention provides a swingable and tiltable suction filter device for CTC enrichment staining, including a fixing component, to avoid the generation of wrinkles and bubbles, and to remove residual reagents in the chip by swinging and tilting.

[0004] To achieve the above object, the present invention provides the following technical solutions: The present invention discloses a suction filtration device for CTC enrichment and staining, comprising a fixing component and a microfluidic chip mounted on the fixing component, wherein the microfluidic chip comprises a reagent tank and a rigid chip fixing plate arranged at the lower end of the reagent tank, wherein a through hole 1 is provided on the rigid chip fixing plate, wherein a chip body is placed in the through hole 1, wherein the reagent tank is a sheet-like structure with a liquid leakage hole longitudinally penetrating the body thereof, and a liquid suction component is arranged on the lower surface of the rigid chip fixing plate.

[0005] Preferably, the fixing component comprises an upper frame and a lower frame arranged parallel to each other, a left frame and a right frame arranged parallel to each other are fixedly connected between the upper frame and the lower frame, rollers are installed at both ends of the upper frame and the lower frame, and upper guide plates and lower guide plates arranged opposite to each other are fixedly connected on the side walls of the upper frame and the lower frame, and the upper guide plates and the lower guide plates are both strip structures with L-shaped cross-sections; the left frame and the right frame are provided with upper guide holes at positions corresponding to the upper guide plates, and lower guide holes at positions corresponding to the lower guide plates; the two ends of the rigid chip fixing plate are overlapped on the upper guide plates and the lower guide plates.

[0006] With the above structural design, the rigid chip is installed between the upper frame and the lower frame, and the residual reagents in the reagent tank and the chip can be removed by swinging and tilting the fixed assembly, which facilitates automated operation.

[0007] Preferably, the chip body is made of a transparent rigid material, and a plurality of groups of long strip-shaped micropores are evenly distributed on the chip body; the micropores on the chip body include transverse holes arranged transversely and vertical holes arranged longitudinally.

[0008] With the above structural design, the rigid chip body can avoid the generation of wrinkles and bubbles during the enrichment and staining process.

[0009] Preferably, the liquid absorption component comprises filter paper adhered to the lower surface of the rigid chip fixing sheet, and a polymer water absorption material layer arranged between the filter paper and the rigid chip fixing sheet.

[0010] The above structural design uses polymer water-absorbing materials to fully absorb various reagents, replacing the previous suction filtration components, reducing the volume of the equipment and reducing costs. It can effectively avoid cell stacking or agglomeration caused by excessive negative pressure during suction filtration.

[0011] Preferably, the polymer water-absorbing material layer of the liquid-absorbing component covers the entire through hole 1, and a gasket is provided between the lower surface of the rigid chip fixing sheet and the liquid-absorbing component, and the gasket surrounds the polymer water-absorbing material layer.

[0012] With the above structural design, the gasket fixes the powdered polymer material within the range of the gasket, preventing the powdered polymer material from being too dispersed and affecting the water absorption effect.

[0013] Preferably, a suction filter bin is detachably mounted at the lower end of the reagent tank via a first card slot, the suction filter bin is a container with an upper opening receiving the liquid leakage hole, and the rigid chip fixing sheet is located between the reagent tank and the suction filter bin.

[0014] With the above structural design, the suction filter bin can conveniently collect the waste reagents and the suction components passing through the leakage hole.

[0015] Preferably, a funnel-shaped liquid collecting groove is provided on the upper surface of the reagent tank, and the lower end of the liquid collecting groove is connected to the liquid leakage hole.

[0016] With the above structural design, a liquid collecting tank is provided to facilitate temporary storage of blood and various reagents added subsequently.

[0017] Preferably, a drainage groove is provided on the side of the upper end of the liquid collecting tank, and the inner end of the drainage groove is connected to the liquid collecting tank, and the outer end of the drainage groove is connected to the outside of the reagent tank.

[0018] The above structural design can facilitate the discharge of the residual reagents in the liquid collecting tank and the liquid leakage hole.

[0019] Preferably, a sealing ring is provided on the lower surface of the reagent tank, the sealing ring coincides with the central axis of the through hole one, and the diameter of the sealing ring is larger than the diameter of the through hole one.

[0020] With the above structural design, a sealing ring is provided between the reagent tank and the rigid chip fixing plate to effectively seal the rigid chip fixing plate and the reagent tank, thereby preventing the subsequent added reagent from overflowing and preventing the reagent from flowing out of the gap between the reagent tank and the rigid chip fixing plate.

[0021] Preferably, the upper guide plate and the lower guide plate are both strip structures with L-shaped cross-sections.

[0022] With the above structural design, the guide plate is light in weight and high in strength, and the L-shaped cross section is groove-shaped after being inclined, and can be used to guide reagents.

[0023] Compared with the prior art, the present invention has the following beneficial effects: The suction filtration device uses a rigid chip with uniform pore size, strong selectivity, little filtrate residue, and no negative pressure suction filtration, which can effectively avoid cell stacking or compaction caused by excessive negative pressure during suction filtration, maintain the original state of the cells as much as possible, and improve the quality of CTC cell enrichment staining; the fixed component is light in weight and strong in strength, and the rigid chip is installed between the upper frame and the lower frame to avoid wrinkles and bubbles. The residual reagents in the chip can be removed by swinging and tilting the fixed component, which facilitates automated operation. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1It is a schematic diagram of the three-dimensional structure of a fixing assembly according to an embodiment of the present invention.

[0025] Figure 2 A schematic diagram of the three-dimensional structure of a reagent tank carrying a microfluidic chip.

[0026] Figure 3 yes Figure 2 Schematic diagram of the cross-sectional structure of the reagent tank shown.

[0027] Figure 4 It is a schematic diagram of the three-dimensional structure of a microfluidic chip.

[0028] Figure 5 It is a schematic diagram of the enlarged structure of some micropores in a microfluidic chip.

[0029] Figure 6 It is a schematic diagram of the three-dimensional structure of the liquid suction component.

[0030] Figure 7 It is a three-dimensional structural schematic diagram of the suction filter bin.

[0031] Figure 8 It is a cross-sectional view of the reagent tank, microfluidic chip and suction filter chamber assembled together. DETAILED DESCRIPTION

[0032] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0033] The suction filtration device for CTC enrichment staining described in the present invention comprises a fixing component and a microfluidic chip mounted on the fixing component. The microfluidic chip comprises a reagent tank 1 and a rigid chip fixing sheet 201 arranged under the reagent tank 1. Figure 1 As shown, the fixed assembly includes an upper frame 501 and a lower frame 502 arranged in parallel with each other, and a left frame 503 and a right frame 504 are fixedly connected between the upper frame 501 and the lower frame 502, which are parallel to each other, and together form a stable rectangular frame structure. Both ends of the upper frame 501 and the lower frame 502 extend out of the left frame 503 and the right frame 504, and rollers 505 are installed at both ends of the upper frame 501 and the lower frame 502, and the axis of the rollers 505 is parallel to the upper frame 501 or the lower frame 502. When in use, the two rollers on the left side can be installed in the same slide groove, and the two rollers on the right side can be installed in another slide groove, so that the fixed assembly can move along the slide groove; in addition, the driving mechanism can be used to lift one of the upper frame 501 and the lower frame 502, so that the fixed assembly can be tilted at a certain angle.

[0034] An upper guide plate 506 and a lower guide plate 507 which are arranged opposite to each other are fixedly connected to the side walls of the upper frame 501 and the lower frame 502. The upper guide plate 506 and the lower guide plate 507 are both strip structures with L-shaped cross sections or U-shaped strip structures.

[0035] To facilitate the installation of the roller 505, the upper frame 501 and the lower frame 502 are both long cylindrical structures, the upper guide plate 506 is welded or fixedly connected to the side of the upper frame 501 by screws, and the lower guide plate 507 is fixedly connected to the side of the lower frame 502, and the upper guide plate 506 and the lower guide plate 507 are slightly shorter and are only located between the left frame 503 and the right frame 504. The two ends of the upper guide plate 506 are respectively connected to the inner upper ends of the left frame 503 and the right frame 504, and the two ends of the lower guide plate 507 are respectively connected to the inner lower ends of the left frame 503 and the right frame 504.

[0036] The left frame 503 and the right frame 504 are provided with upper guide holes 508 at positions corresponding to the upper guide plate 506, and lower guide holes 509 at positions corresponding to the lower guide plate 507. The upper guide holes 508 and the lower guide holes 509 are through holes provided on the left frame 503 and the right frame 504, and are used to guide the test reagents inside the left frame 503 and the right frame 504 to the outside, and then collect the test reagents through other guide devices for centralized disposal.

[0037] In addition, to improve strength and reduce weight, the side walls of the left frame 503 and the right frame 504 are fixedly connected with the left frame folding edge and the right frame folding edge 514 arranged opposite to each other. The left frame 503 and the left frame folding edge, the right frame 504 and the right frame folding edge 514 form a right-angle folding structure, which has greater strength.

[0038] like Figure 1 As shown, when in use, the rigid chip fixing sheet 201 is attached to the bottom of the reagent tank 1. The rigid chip fixing sheet 2 is in the form of a glass slide, and the rigid chip fixing sheet 201 and the reagent tank 1 are clamped between the upper frame 501 and the lower frame 502. The width of the reagent tank 1 is smaller than the spacing between the left frame 503 and the right frame 504, and one reagent tank 1 can be placed between the left frame 503 and the right frame 504, or multiple reagent tanks 1 can be placed in parallel at the same time.

[0039] like Figure 2 , Figure 3 As shown, the upper surface of the reagent tank 1 is provided with a liquid collecting groove 106, and the liquid collecting groove 106 is a funnel-shaped structure with a large upper portion and a small lower portion. The lower end of the liquid collecting groove 106 is connected to a liquid leakage hole 102, and the side of the upper end of the liquid collecting groove 106 is provided with a drainage groove 105, the inner end of the drainage groove 105 is connected to the liquid collecting groove 106, and the outer end is connected to the outside of the reagent tank 1. The lower end of the reagent tank 1 is detachably mounted with a suction filter bin 4 through a first card slot 103, as shown in FIG. Figure 7 As shown, the suction filter chamber 4 is a container with an upper opening and receiving the leakage hole 102; the rigid chip fixing plate 201 is located between the reagent tank 1 and the suction filter chamber 4; a through hole 202 is opened on the rigid chip fixing plate 201, and a chip body 2 is placed in the through hole 202.

[0040] When the reagent tank 1 is placed horizontally, the liquid such as the reagent that drops through the liquid leakage hole 102 passes through the chip body 2 and is collected in the suction filter chamber 4 .

[0041] like Figure 4 , Figure 5 As shown, the material of the chip body 2 is preferably flexible glass, and a plurality of groups of long strip-shaped micropores are evenly distributed on the chip body 2; the micropores include a transverse hole 11 arranged horizontally and a vertical hole 12 arranged longitudinally. The width of the transverse hole 11 and the vertical hole 12 are both 0.0075mm, the length of the transverse hole 11 is 0.09mm, and the length of the vertical hole 12 is 0.06mm. The transverse hole 11 and the vertical hole 12 are arranged perpendicular to each other, the spacing between adjacent transverse holes 11 is 0.03mm, and the vertical hole 12 is located on the center line of the transverse hole 11. The spacing between the transverse holes and the upper ends of the vertical holes in the same hole group is 0.015mm. The spacing between adjacent vertical holes is large, which can effectively avoid cell superposition.

[0042] like Figure 6 , Figure 8 As shown, a liquid absorption component 3 is arranged on the lower surface of the rigid chip fixing plate 201 , and the liquid absorption component 3 includes a filter paper 301 adhered to the rigid chip fixing plate 201 and a polymer water absorption material layer 303 arranged between the filter paper 301 and the microfluidic chip 1 .

[0043] The polymer water-absorbing material layer 303 uses a powdered super absorbent resin, which is a new type of functional polymer material. It has a high water absorption function of absorbing water that is hundreds to thousands of times heavier than itself, and has excellent water retention performance. Once it absorbs water and swells into a hydrogel, it is difficult to separate the water even under pressure. Super absorbent resin is generally a polymer electrolyte containing hydrophilic groups and cross-linked structures. Before absorbing water, the polymer chains are entangled with each other and cross-linked to form a network structure, thereby achieving overall tightening. When in contact with water, water molecules penetrate into the resin through capillary action and diffusion, and the ionized groups on the chain are ionized in water. Due to the electrostatic repulsion between the same ions on the chain, the polymer chain stretches and swells. Due to the requirement of electrical neutrality, the counterions cannot migrate to the outside of the resin, and the ion concentration difference between the solution inside and outside the resin forms a reverse osmotic pressure. Under the action of reverse osmotic pressure, water further enters the resin to form a hydrogel.

[0044] like Figure 6 , Figure 8As shown, a gasket 302 is provided between the lower surface of the rigid chip fixing plate 201 and the liquid absorption component 3, and the gasket 302 surrounds the polymer water-absorbing material layer 303. The function of the gasket 302 is to fix the powdered polymer material within the range of the gasket 302 to prevent the powdered polymer material from being too dispersed and affecting the water absorption effect. The filter paper 301 can be used in multiple layers. When in use, the layer of filter paper 301 farthest from the chip body 2 is fixed to the lower bottom surface of the rigid chip fixing plate 201. After the polymer water-absorbing material layer 303 absorbs enough reagents, the volume expands, causing the liquid absorption component 3 to detach and fall into the suction filter bin 4 under the action of gravity.

[0045] like Figure 2 , Figure 3 As shown, in this embodiment, drainage grooves 105 are provided at two opposite corners along the length direction of the reagent tank 1, and the drainage grooves 105 are located on both sides of the reagent tank 1 facing the upper frame 501 or the lower frame 502, and the inner end of the drainage groove 105 is connected to the chip fixing groove 101, and the outer end is connected to the outside of the reagent tank 1. In this way, after the cell staining is completed, the reagent tank 1 can be tilted by raising the upper frame 501 or the lower frame 502, and the excess reagent in the leakage hole 102 can be concentratedly drained to the outside of the reagent tank 1 through the drainage grooves 105, and flow along the upper guide plate 506 or the lower guide plate 507 to the outside of the left frame 503 and the right frame 504, which is conducive to the centralized disposal of waste reagents.

[0046] like Figure 7 , Figure 8 As shown, the lower end of the reagent tank 1 is detachably mounted with a suction filter bin 4 through a first card slot 103. The suction filter bin 4 is a container with an upper opening and receiving the leakage hole 102. The edge of the upper port of the suction filter bin 4 is provided with a claw 401 matching the first card slot 103. When in use, the suction filter bin 4 can be installed to the lower end of the reagent tank 1 by simply inserting the claw 401 into the first card slot 103. After installation, the upper end opening of the suction filter bin 4 is opposite to the leakage hole 102, and the waste reagent flowing out from the leakage hole 102 through the chip body 2 and the liquid absorption component 3 falling off the chip body 2 are temporarily stored in the suction filter bin 4. After the cell enrichment process is completed, since the volume and weight of the polymer water-absorbing material increase significantly after absorbing water, the suction filter bin can prevent the polymer material from overflowing after absorbing water and reduce pollution to the surrounding environment. Precisely because the volume and weight of the polymer water-absorbing material increase significantly after absorbing water, the liquid absorption component 3 automatically falls off from the chip fixing groove under the action of its gravity, eliminating the manual peeling step, reducing labor and avoiding pollution.

[0047] like Figure 8As shown, when in use, the rigid chip fixing sheet 201 is fitted under the reagent tank 1, the liquid absorption component 3 is located under the chip body 2, and the suction filter chamber 4 is installed under the reagent tank 1. Blood or reagent is poured into the reagent tank 1, collected in the liquid collecting tank 106, and then passes through the leakage hole 102 and the chip body 2. The blood or reagent passing through the chip body 2 is absorbed by the liquid absorption component 3. The liquid absorption component contains a polymer water-absorbing material, and its volume expands hundreds of times after absorbing water. The liquid absorption component 3 is separated from the rigid chip fixing sheet 201 after absorbing liquid. The liquid absorption component 3 that fully absorbs the reagent will eventually fall off the rigid chip fixing sheet 201 and fall into the suction filter chamber 4. This is the cell enrichment process. Subsequently, at least one dyeing reagent is poured into the reagent tank 1. After dyeing with one dyeing reagent is completed, the upper frame 501 or the lower frame 502 is lifted by the driving device, and one of the upper guide plate 506 or the lower guide plate 507 is lifted accordingly. At this time, the reagent tank 1 is tilted, and the reagent in the liquid collecting tank 106 flows along the lower drainage groove 105 into the lower upper guide plate 506 or the lower guide plate 507, and finally flows along the guide plate to the outside of the left frame 503 or the right frame 504. Another dyeing reagent is poured into the reagent tank 1, and the above dyeing operation is repeated. The reagents in the liquid collecting tank 106 are finally collected and disposed of.

[0048] The contents not described in detail in this specification belong to the prior art known to the professional and technical personnel in this field. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for the technical solutions described in the aforementioned embodiments to be modified, or some of the technical features thereof to be replaced by equivalents, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A suction filtration device for CTC enrichment and staining, comprising a fixing component and a microfluidic chip mounted on the fixing component, characterized in that: The microfluidic chip comprises a reagent tank (1), and a rigid chip fixing plate (201) arranged at the lower end of the reagent tank (1); a through hole (202) is provided on the rigid chip fixing plate (201); a chip body (2) is placed in the through hole (202); the reagent tank (1) is a sheet-like structure with a liquid leakage hole (102) longitudinally penetrating the body; and a liquid aspiration component (3) is arranged on the lower surface of the rigid chip fixing plate (201).

2. A suction filtration device for CTC enrichment and staining according to claim 1, characterized in that: The fixing assembly comprises an upper frame (501) and a lower frame (502) which are arranged in parallel with each other, a left frame (503) and a right frame (504) which are arranged in parallel with each other are fixedly connected between the upper frame (501) and the lower frame (502), rollers (505) are installed at both ends of the upper frame (501) and the lower frame (502), an upper guide plate (506) and a lower guide plate (507) which are arranged opposite to each other are fixedly connected on the side walls of the upper frame (501) and the lower frame (502), an upper guide hole (508) is arranged on the left frame (503) and the right frame (504) at a position corresponding to the upper guide plate (506), and a lower guide hole (509) is arranged on a position corresponding to the lower guide plate (507); and both ends of the rigid chip fixing plate (201) are overlapped on the upper guide plate (506) and the lower guide plate (507).

3. A suction filtration device for CTC enrichment and staining according to claim 1, characterized in that: The chip body (2) is made of a transparent rigid material, and a plurality of groups of long strip-shaped micropores are evenly distributed on the chip body (2); the micropores on the chip body (2) include transverse holes (11) arranged transversely and vertical holes (12) arranged longitudinally.

4. A suction filtration device for CTC enrichment and staining according to claim 1, characterized in that: The liquid absorption component (3) comprises a filter paper (301) adhered to the lower surface of the rigid chip fixing plate (201), and a polymer water absorption material layer (303) arranged between the filter paper (301) and the rigid chip fixing plate (201).

5. A suction filtration device for CTC enrichment and staining according to claim 4, characterized in that: The polymer water-absorbing material layer (303) of the liquid-absorbing component (3) covers the entire through hole 1 (202); a gasket (302) is provided between the lower surface of the rigid chip fixing plate (201) and the liquid-absorbing component (3); and the gasket (302) surrounds the polymer water-absorbing material layer (303).

6. A suction filtration device for CTC enrichment and staining according to claim 1, characterized in that: A suction filter bin (4) is detachably mounted at the lower end of the reagent tank (1) via a first card slot (103); the suction filter bin (4) is a container with an upper opening and receiving the liquid leakage hole (102); and the rigid chip fixing plate (201) is located between the reagent tank (1) and the suction filter bin (4).

7. A suction filtration device for CTC enrichment and staining according to claim 1, characterized in that: A funnel-shaped liquid collecting groove (106) is provided on the upper surface of the reagent tank (1), and the lower end of the liquid collecting groove (106) is connected to the liquid leakage hole (102).

8. A suction filtration device for CTC enrichment and staining according to claim 7, characterized in that: A drainage groove (105) is provided on the side of the upper end of the liquid collecting groove (106), and the inner end of the drainage groove (105) is connected to the liquid collecting groove (106), and the outer end is connected to the outside of the reagent tank (1).

9. A suction filtration device for CTC enrichment and staining according to claim 1, characterized in that: A sealing ring (107) is provided on the lower surface of the reagent tank (1), the sealing ring (107) coincides with the central axis of the through hole one (202), and the diameter of the sealing ring (107) is greater than the diameter of the through hole one (202).

10. A suction filtration device for CTC enrichment and staining according to claim 2, characterized in that: The upper guide plate (506) and the lower guide plate (507) are both strip structures with L-shaped cross-sections.

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