A platelet detection microfluidic chip

Abstract

The present invention discloses a platelet detection microfluidic chip, including a substrate and a cover plate, wherein the cover plate and the substrate are sealed and matched to form a chip body; a plurality of separation detection units are arranged on the chip body, and the plurality of separation detection units are radially distributed with the center of the chip body as the origin; the separation detection unit includes a primary PRP separation module, a secondary PPP separation module and a tertiary detection module arranged on the substrate; the primary separation module is arranged at the center of the chip body, and the secondary separation module and the tertiary detection module are arranged in sequence in a direction away from the center of the chip body. The present invention integrates PRP, PPP separation and detection, achieves the purpose of rapid separation and detection of multiple samples, high detection efficiency, strong reliability and accurate detection effect, and can be used in multiple fields such as molecular detection, medical diagnosis and clinical application.

Description

Technical Field

[0001] The invention relates to the technical field of molecular detection and microfluidic chips, and more particularly to a platelet detection microfluidic chip. Background Art

[0002] In recent years, the separation and detection of plasma has been widely studied in order to ensure public health, disease prevention and beauty care. Current research is tending to separate and detect PRP (platelet-rich plasma) and PPP (platelet-poor plasma). Both PRP and PPP are separated and extracted from whole blood, but the preparation methods and functions are very different.

[0003] The traditional method of separating PRP and PPP is generally to control the centrifugal speed and obtain the required part through stratification after blood centrifugation. The separation and extraction steps are cumbersome, which places high demands on the stability of the centrifugal equipment and the professionalism of the testing technicians.

[0004] In addition, in traditional blood sample extraction and testing, the sample demand is large, the testing cycle is long, and blood samples are not suitable for long-term storage. At room temperature, it is best to separate them within 2 hours. If they are stored for too long, red blood cells may rupture and protein may precipitate, affecting the test results. PRP and PPP need to be separated and tested separately during preparation and testing, which undoubtedly increases the storage time and inactivation risk of the samples.

[0005] Therefore, how to provide a platelet detection microfluidic chip is an urgent problem to be solved by those skilled in the art. Summary of the invention

[0006] In view of this, the present invention provides a platelet detection microfluidic chip to at least solve one of the problems raised in the above background technology section.

[0007] In order to realize the above scheme, the present invention adopts the following technical scheme:

[0008] A platelet detection microfluidic chip comprises a substrate and a cover plate, wherein the cover plate and the substrate are sealed together to form a chip body; a plurality of separation detection units are arranged on the chip body, and the plurality of separation detection units are radially distributed with the center of the chip body as the origin; the separation detection units comprise a primary PRP separation module, a secondary PPP separation module and a tertiary detection module arranged on the substrate; the primary separation module is arranged near the center of the chip body, and the secondary separation module and the tertiary detection module are arranged in sequence in a direction away from the center of the chip body.

[0009] Preferably, in the above-mentioned platelet detection microfluidic chip, the first-level PRP separation module includes a whole blood sampling pool, a plasma separation pool, a PRP storage pool and a blood cell storage pool, the whole blood sampling pool is arranged at the center of the circle close to the chip body, and the whole blood sampling pool, the plasma separation pool and the blood cell storage pool are arranged in sequence along the direction away from the center of the circle of the chip body, and the PRP storage pool is arranged on one side of the blood cell storage pool; the whole blood sampling pool is connected to the plasma separation pool through a first channel, the plasma separation pool is connected to the blood cell storage pool through a second channel, and the plasma separation pool and the blood cell storage pool are also connected through a connecting channel; the plasma separation pool is connected through a first capillary siphon channel; and a PRP storage pool exhaust hole is provided on the PRP storage pool.

[0010] Preferably, in the above-mentioned platelet detection microfluidic chip, the secondary PPP separation module includes a liquid separation channel, a waste liquid pool, a PRP quantitative pool, a quantitative separation pool, a sedimentation pool and a PPP quantitative pool; the waste liquid pool, the PRP quantitative pool and the quantitative separation pool are arranged on the side of the PRP storage pool away from the center of the chip body, and the PRP quantitative pool and the quantitative separation pool are connected with the PRP storage pool through a liquid separation channel; the waste liquid pool is connected with the liquid separation channel through a third channel; the quantitative separation pool is connected with the sedimentation pool on the side away from the center of the chip body; the quantitative separation pool is connected with the PPP quantitative pool through a second capillary siphon channel; the quantitative separation pool is provided with a quantitative separation pool exhaust hole, and the PPP quantitative pool is provided with a PPP quantitative pool exhaust hole.

[0011] Preferably, in the above-mentioned platelet detection microfluidic chip, the three-stage detection module includes a first reagent injection pool, a second reagent injection pool, a first detection pool and a second detection pool; the first detection pool is arranged on the side of the PRP quantitative pool away from the center of the chip body, and is connected with the PRP quantitative pool through the fourth channel, and the fourth channel is provided with a first microfluidic valve; the first reagent injection pool is arranged on the side of the first detection pool close to the center of the chip body, and is connected with the first detection pool through the fifth channel; the second detection pool is arranged on the side of the PPP quantitative pool away from the center of the chip body, and is connected with the PPP quantitative pool through the sixth channel, and the sixth channel is provided with a second microfluidic valve; the second reagent injection pool is arranged on the side of the second detection pool close to the center of the chip body, and is connected with the second detection pool through the seventh channel; the first reagent injection pool is provided with a first reagent injection pool exhaust hole, and the second reagent injection pool is provided with a second reagent injection pool exhaust hole.

[0012] Preferably, in the above-mentioned platelet detection microfluidic chip, the cover plate is provided with a whole blood injection hole, a whole blood injection pool air hole, a PRP storage pool air hole, a quantitative separation pool air hole, a PPP quantitative pool air hole, a first reagent injection pool air hole, and a second reagent injection pool air hole; the whole blood injection hole and the whole blood injection pool air hole correspond to and are connected with the whole blood injection pool; the PRP storage pool air hole corresponds to and is connected with the PRP storage pool exhaust hole; the quantitative separation pool air hole corresponds to and is connected with the quantitative separation pool exhaust hole; the PPP quantitative pool air hole corresponds to and is connected with the PPP quantitative pool exhaust hole; the first reagent injection pool air hole corresponds to and is connected with the first reagent injection pool exhaust hole; the second reagent injection pool air hole corresponds to and is connected with the second reagent injection pool exhaust hole. The above-mentioned hole structure can ensure the smooth flow of air inside the chip and promote liquid circulation.

[0013] Preferably, in the above-mentioned platelet detection microfluidic chip, the whole blood sampling pool, plasma separation pool, PRP storage pool, blood cell storage pool, liquid separation channel, waste liquid pool, PRP quantitative pool, quantitative separation pool, sedimentation pool, PPP quantitative pool, first reagent sampling pool, second reagent sampling pool, first detection pool, second detection pool, first capillary siphon channel, second capillary siphon channel, connecting channel, first channel, second channel, third channel, fourth channel, fifth channel, sixth channel, and seventh channel are groove structures formed on the side of the substrate facing the cover plate by etching or cutting. The above-mentioned various channels facilitate the circulation and separation of liquids under the action of centrifugal force.

[0014] Preferably, in the above-mentioned platelet detection microfluidic chip, the whole blood sampling pool is in a hook shape, and the whole blood sampling hole corresponds to and is connected to the longer end of the whole blood sampling pool, and the whole blood sampling pool air hole corresponds to and is connected to the shorter end of the whole blood sampling pool.

[0015] Preferably, in the above-mentioned platelet detection microfluidic chip, a cover fixing hole is provided at the center of the cover plate, and a substrate fixing hole is provided at the center of the substrate; the cover fixing hole and the substrate fixing hole have the same shape and are arranged to overlap, forming a chip fixing hole that passes through the chip body.

[0016] Preferably, in the above-mentioned platelet detection microfluidic chip, the cover plate and the substrate are made of one of silicon wafer, quartz, glass and polymer compound.

[0017] Preferably, in the above-mentioned platelet detection microfluidic chip, the polymer compound is polymethacrylate or polystyrene or cycloolefin copolymer or polycarbonate.

[0018] It can be seen from the above technical solutions that, compared with the prior art, the present invention discloses a platelet detection microfluidic chip, which integrates PRP and PPP separation and detection, achieves the purpose of rapid separation and detection of multiple samples, high detection efficiency, strong reliability and accurate detection effect, and can be used in multiple fields such as molecular detection, medical diagnosis and clinical application. The present invention has the following beneficial effects:

[0019] 1. The platelet detection microfluidic chip of the present invention can realize batch detection and quick use, greatly reduce the risk of red blood cell rupture and protein denaturation, and improve separation efficiency and detection accuracy.

[0020] 2. The amount of sample extracted in blood sample detection is limited. The microfluidic chip in the present invention can realize the one-time separation and simultaneous detection of PRP and PPP, reduce the amount of blood sample, improve the utilization rate of blood sample, and rationalize the separation and detection process.

[0021] 3. The chip is designed with multiple modules including a primary PRP separation module, a secondary PPP separation module and a tertiary detection module. The modules and channels are reasonably designed so that two tests can be performed separately and simultaneously, which can effectively avoid the backflow and contamination of liquid samples.

[0022] 4. The microfluidic chip of the present invention is easy to operate and can be programmed to separate PRP and PPP samples at one time, overcoming the traditional multi-step separation and complex manual operation.

[0023] 5. It realizes the integration of PRP and PPP separation and detection, integrates the injection, processing and detection modules, reduces the influence of external factors, and achieves the effect of fast speed, small volume and accurate results. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0025] Figure 1 The accompanying drawing is a schematic diagram of the structure of the present invention;

[0026] Figure 2 The accompanying drawing is a schematic structural diagram of the separation and detection unit. DETAILED DESCRIPTION

[0027] 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.

[0028] The embodiment of the present invention discloses a platelet detection microfluidic chip, comprising a substrate and a cover plate, wherein the cover plate and the substrate are sealed and matched to form a chip body 1; a plurality of separation detection units 2 are arranged on the chip body 1, and the plurality of separation detection units 2 are radially distributed with the center of the chip body 1 as the origin; the separation detection unit 2 comprises a primary PRP separation module, a secondary PPP separation module and a tertiary detection module arranged on the substrate; the primary separation module is arranged at the center of the chip body 1, and the secondary separation module and the tertiary detection module are arranged in sequence in a direction away from the center of the chip body 1.

[0029] In order to further optimize the above technical scheme, the first-level PRP separation module includes a whole blood sampling pool 3, a plasma separation pool 4, a PRP storage pool 5 and a blood cell storage pool 6. The whole blood sampling pool 3 is arranged at the center of the circle close to the chip body 1, and the whole blood sampling pool 3, the plasma separation pool 4, and the blood cell storage pool 6 are arranged in sequence along the direction away from the center of the circle of the chip body 1, and the PRP storage pool 5 is arranged on one side of the blood cell storage pool 6; the whole blood sampling pool 3 is connected to the plasma separation pool 4 through the first channel 7, the plasma separation pool 4 is connected to the blood cell storage pool 6 through the second channel 8, and the plasma separation pool 4 and the blood cell storage pool 6 are also connected through a connecting channel 9; the plasma separation pool 4 is connected through the first capillary siphon channel 10; and the PRP storage pool 5 is provided with a PRP storage pool exhaust hole 11.

[0030] In order to further optimize the above technical scheme, the secondary PPP separation module includes a liquid separation channel 12, a waste liquid pool 13, a PRP quantitative pool 14, a quantitative separation pool 15, a sedimentation pool 16 and a PPP quantitative pool 17; the waste liquid pool 13, the PRP quantitative pool 14 and the quantitative separation pool 15 are arranged on the side of the PRP storage pool 5 away from the center of the chip body 1, and the PRP quantitative pool 14 and the quantitative separation pool 15 are connected with the PRP storage pool 5 through the liquid separation channel 12; the waste liquid pool 13 is connected with the liquid separation channel 12 through the third channel 18; the quantitative separation pool 15 is connected with the sedimentation pool 16 on the side away from the center of the chip body 1; the quantitative separation pool 15 is connected with the PPP quantitative pool 17 through the second capillary siphon channel 19; the quantitative separation pool 15 is provided with a quantitative separation pool exhaust hole 20, and the PPP quantitative pool 17 is provided with a PPP quantitative pool exhaust hole 21.

[0031] In order to further optimize the above technical scheme, the three-level detection module includes a first reagent injection pool 22, a second reagent injection pool 23, a first detection pool 24 and a second detection pool 25; the first detection pool 24 is arranged on the side of the PRP quantitative pool 14 away from the center of the chip body 1, and is connected to the PRP quantitative pool 14 through the fourth channel 26, and a first microfluidic valve 27 is arranged on the fourth channel 26; the first reagent injection pool 22 is arranged on the side of the first detection pool 24 close to the center of the chip body 1, and is connected to the first detection pool 24 through the fifth channel 28; the second detection pool 25 is arranged on the side of the PPP quantitative pool 17 away from the center of the chip body 1, and is connected to the PPP quantitative pool 17 through the sixth channel 29, and a second microfluidic valve 30 is arranged on the sixth channel 29; the second reagent injection pool 23 is arranged on the side of the second detection pool 25 close to the center of the chip body 1, and is connected to the second detection pool 25 through the seventh channel 31; a first reagent injection pool exhaust hole 32 is arranged on the first reagent injection pool 22, and a second reagent injection pool exhaust hole 33 is arranged on the second reagent injection pool 23.

[0032] In order to further optimize the above technical scheme, the cover plate is provided with a whole blood injection hole, a whole blood injection pool air hole, a PRP storage pool air hole, a quantitative separation pool air hole, a PPP quantitative pool air hole, a first reagent injection pool air hole, and a second reagent injection pool air hole; the whole blood injection hole and the whole blood injection pool air hole correspond to and are connected with the whole blood injection pool 3; the PRP storage pool air hole corresponds to and is connected with the PRP storage pool exhaust hole 11; the quantitative separation pool air hole corresponds to and is connected with the quantitative separation pool exhaust hole 20; the PPP quantitative pool air hole corresponds to and is connected with the PPP quantitative pool exhaust hole 21; the first reagent injection pool air hole corresponds to and is connected with the first reagent injection pool exhaust hole 32; the second reagent injection pool air hole corresponds to and is connected with the second reagent injection pool exhaust hole 33. The above hole structure can ensure the smooth flow of air inside the chip and promote liquid circulation.

[0033] In order to further optimize the above technical solution, the whole blood injection pool 3, the plasma separation pool 4, the PRP storage pool 5, the blood cell storage pool 6, the liquid separation channel 12, the waste liquid pool 13, the PRP quantitative pool 14, the quantitative separation pool 15, the sedimentation pool 16, the PPP quantitative pool 17, the first reagent injection pool 22, the second reagent injection pool 23, the first detection pool 24, the second detection pool 25, the first capillary siphon channel 10, the second capillary siphon channel 19, the connecting channel 9, the first channel 7, the second channel 8, the third channel 18, the fourth channel 26, the fifth channel 28, the sixth channel 29, and the seventh channel 31 are groove structures formed on the side of the substrate facing the cover plate by etching or cutting. The above-mentioned various channels facilitate the circulation and separation of liquids under the action of centrifugal force.

[0034] In order to further optimize the above technical solution, the whole blood sampling pool 3 is hook-shaped, and the whole blood sampling hole corresponds to and is connected to the longer end of the whole blood sampling pool 3, and the whole blood sampling pool vent hole corresponds to and is connected to the shorter end of the whole blood sampling pool 3.

[0035] In order to further optimize the above technical solution, a cover fixing hole is set at the center of the cover plate, and a substrate fixing hole is set at the center of the substrate; the cover fixing hole and the substrate fixing hole are of the same shape and overlap with each other to form a chip fixing hole 34 that passes through the chip body 1. The chip fixing hole 34 is used to cooperate with the centrifugal rotating equipment for installation to rotate the chip. Different plasma samples can be obtained by adjusting the centrifugal force and the centrifugal time.

[0036] In order to further optimize the above technical solution, the material of the cover plate and the substrate is one of silicon wafer, quartz, glass, and polymer compound.

[0037] In order to further optimize the above technical solution, the polymer compound is polymethacrylate or polystyrene or cycloolefin copolymer or polycarbonate.

[0038] Detection method:

[0039] The sample separation and detection process is as follows: first, add the whole blood sample to be tested from the whole blood injection hole, rotate at a clockwise rate of 3000-4500r / min for 60-180s, and the sample enters the plasma separation pool 4 and the blood cell storage pool 6 respectively. The rotation stops and waits for 5-20s, and the plasma enters the first capillary siphon channel 10 through siphon action, and the liquid enters the PRP storage pool 5 at a counterclockwise rate of 600-1000r / min for 30-60s. The collected liquid is PRP, and the primary separation is completed;

[0040] Then, the operation is carried out at 4000-5500r / min, either clockwise or reverse, and after 2-3min, PRP enters the quantitative separation tank 15, and a part of PRP is transferred to the first detection tank 24 after quantification, and the other part is centrifuged to produce PPP and precipitate, and the precipitate enters the sedimentation tank 16. After another 30-60s of clockwise rotation at 500-1000r / min, PPP is transferred to the PPP quantitative tank 17, and the secondary separation is completed;

[0041] Then, PPP is transferred into the second detection pool 25 at 2000r / min for 5-10s, and PRP and PPP detection reagents are added into the first reagent injection pool 22 and the second reagent injection pool 23 respectively. The transfer and mixing of PRP and PPP with the detection reagents are completed at 2000r / min for 5-10s, followed by reaction and detection, thereby completing the tertiary detection.

[0042] In this specification, each embodiment is described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the embodiments can be referred to each other. For the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and the relevant parts can be referred to the method part.

[0043] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A platelet detection microfluidic chip, comprising a substrate and a cover plate, wherein the cover plate and the substrate are sealed to form a chip body; characterized in that: The chip body is provided with a plurality of separation and detection units, which are radially distributed with the center of the chip body as the origin; the separation and detection units include a primary PRP separation module, a secondary PPP separation module and a tertiary detection module arranged on the substrate; the primary PRP separation module is arranged at the center of the chip body, and the secondary PPP separation module and the tertiary detection module are arranged in sequence in a direction away from the center of the chip body; The primary PRP separation module includes a whole blood sampling pool, a plasma separation pool, a PRP storage pool and a blood cell storage pool, the whole blood sampling pool is arranged at the center of the chip body, and the whole blood sampling pool, the plasma separation pool and the blood cell storage pool are arranged in sequence in a direction away from the center of the chip body, and the PRP storage pool is arranged on one side of the blood cell storage pool; the whole blood sampling pool is connected to the plasma separation pool through a first channel, the plasma separation pool is connected to the blood cell storage pool through a second channel, and the plasma separation pool and the blood cell storage pool are also connected through a connecting channel; the plasma separation pool is connected through a first capillary siphon channel; the PRP storage pool is provided with a PRP storage pool exhaust hole; The secondary PPP separation module includes a liquid separation channel, a waste liquid pool, a PRP quantitative pool, a quantitative separation pool, a precipitation pool and a PPP quantitative pool; the waste liquid pool, the PRP quantitative pool and the quantitative separation pool are arranged on the side of the PRP storage pool away from the center of the chip body, and the PRP quantitative pool and the quantitative separation pool are connected with the PRP storage pool through a liquid separation channel; the waste liquid pool is connected with the liquid separation channel through a third channel; the quantitative separation pool is connected with the precipitation pool on the side away from the center of the chip body; the quantitative separation pool is connected with the PPP quantitative pool through a second capillary siphon channel; the quantitative separation pool is provided with a quantitative separation pool exhaust hole, and the PPP quantitative pool is provided with a PPP quantitative pool exhaust hole; The three-level detection module includes a first reagent injection pool, a second reagent injection pool, a first detection pool and a second detection pool; the first detection pool is arranged on the side of the PRP quantitative pool away from the center of the chip body, and is connected to the PRP quantitative pool through a fourth channel, and a first microfluidic valve is arranged on the fourth channel; the first reagent injection pool is arranged on the side of the first detection pool close to the center of the chip body, and is connected to the first detection pool through a fifth channel; the second detection pool is arranged on the side of the PPP quantitative pool away from the center of the chip body, and is connected to the PPP quantitative pool through a sixth channel, and a second microfluidic valve is arranged on the sixth channel; the second reagent injection pool is arranged on the side of the second detection pool close to the center of the chip body, and is connected to the second detection pool through a seventh channel; the first reagent injection pool is provided with a first reagent injection pool exhaust hole, and the second reagent injection pool is provided with a second reagent injection pool exhaust hole.

2. A platelet detection microfluidic chip according to claim 1, characterized in that: The cover plate is provided with a whole blood sampling hole, a whole blood sampling pool air hole, a PRP storage pool air hole, a quantitative separation pool air hole, a PPP quantitative pool air hole, a first reagent sampling pool air hole, and a second reagent sampling pool air hole; the whole blood sampling hole and the whole blood sampling pool air hole correspond to and are connected with the whole blood sampling pool; the PRP storage pool air hole corresponds to and is connected with the PRP storage pool exhaust hole; the quantitative separation pool air hole corresponds to and is connected with the quantitative separation pool exhaust hole; the PPP quantitative pool air hole corresponds to and is connected with the PPP quantitative pool exhaust hole; the first reagent sampling pool air hole corresponds to and is connected with the first reagent sampling pool exhaust hole; the second reagent sampling pool air hole corresponds to and is connected with the second reagent sampling pool exhaust hole.

3. A platelet detection microfluidic chip according to claim 2, characterized in that: The whole blood injection pool, plasma separation pool, PRP storage pool, blood cell storage pool, liquid separation channel, waste liquid pool, PRP quantitative pool, quantitative separation pool, sedimentation pool, PPP quantitative pool, first reagent injection pool, second reagent injection pool, first detection pool, second detection pool, first capillary siphon channel, second capillary siphon channel, connecting channel, first channel, second channel, third channel, fourth channel, fifth channel, sixth channel, and seventh channel are groove structures.

4. A platelet detection microfluidic chip according to claim 2, characterized in that: The whole blood sampling pool is hook-shaped, and the whole blood sampling hole corresponds to and is connected with a longer end of the whole blood sampling pool, and the whole blood sampling pool air permeability hole corresponds to and is connected with a shorter end of the whole blood sampling pool.

5. The platelet detection microfluidic chip according to claim 1, characterized in that: A cover plate fixing hole is arranged at the center of the cover plate, and a substrate fixing hole is arranged at the center of the substrate; the cover plate fixing hole and the substrate fixing hole have the same shape and are arranged to overlap, forming a chip fixing hole that penetrates the chip body.

6. A platelet detection microfluidic chip according to claim 1, characterized in that: The cover plate and the substrate are made of one of silicon wafer, quartz, glass and polymer compound.

7. A platelet detection microfluidic chip according to claim 6, characterized in that: The polymer compound is polymethacrylate or polystyrene or cycloolefin copolymer or polycarbonate.

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