High-throughput chip for detecting platelet function and medical device thereof

Abstract

The application discloses a high-throughput chip for detecting platelet function, which comprises a base layer, a reaction layer arranged on the base layer and a flow layer arranged between the base layer and the reaction layer for accommodating fluid. A liquid guide port is connected with a fluid distribution device for inputting solution droplets and power fluid into the fluid distribution device. The fluid distribution device is arranged at the outer edge of the flow layer and comprises at least one distribution port for inputting the solution droplets and the power fluid into the flow layer and uniformly distributing the solution droplets and the power fluid. The reaction layer comprises a plurality of reaction cavities, each of the reaction cavities comprises a first accommodating cavity and a second accommodating cavity, and the accommodating space of the first accommodating cavity is larger than that of the second accommodating cavity. The high-throughput chip can effectively control the position, size and quantity of the droplets, and can realize fusion of a plurality of different types of droplets, so that high-throughput reaction can be realized in a micro volume, the volume requirement of the platelet-rich plasma to be detected is reduced, and the observation and analysis of the platelet aggregation condition are facilitated, and a new technical scheme is provided for platelet function research.

Description

Technical Field

[0001] This invention belongs to the field of biological detection technology, and in particular relates to a chip for high-throughput detection of numerous trace platelet samples to examine platelet aggregation function, which is beneficial for platelet activation research and drug screening, as well as its application as a medical device. Background Technology

[0002] Platelets are anucleate cell fragments produced by megakaryocytes and participate in numerous physiological and pathological processes, especially the formation of arterial thrombosis. Under physiological conditions, platelet activation is maintained within a controllable range, playing a physiological hemostatic role. However, under certain pathological conditions, excessive platelet activation can lead to arterial thrombosis. Therefore, platelets are crucial for arterial thrombosis, and in-depth research into the regulatory mechanisms of platelet activation is of great value for the prevention and treatment of arterial thrombotic diseases.

[0003] In normal physiological processes, platelets undergo activation, adhesion, release, aggregation, and contraction to fulfill their functions, and each step is strictly regulated by different signaling pathways. Platelet aggregation is a crucial component in pathological conditions such as arterial thrombosis and thrombosis on the surface of intravascular implants like stents, and evaluating platelet aggregation characteristics is the most fundamental test in platelet research.

[0004] CN103558152A discloses a rapid whole blood platelet aggregation function detection device. This device utilizes low-speed centrifugation to allow red blood cells, white blood cells, and platelet aggregation complexes to settle to the bottom of the detection tube. Unaggregated platelet monomers remain in the supernatant (plasma). The supernatant is then directly irradiated with visible light. Since platelets absorb and scatter visible light, the absorption and scattering of light by platelets at a specific wavelength are related to the platelet concentration for the same volume of platelets. By detecting the transmitted light signal or the intensity of the scattered light in the supernatant under specific wavelength irradiation and comparing it with the transmitted or scattered light signals of the corresponding control tube, the platelet content in the supernatant can be detected. Based on this, the platelet aggregation rate can be calculated, thus reflecting the platelet aggregation function level of the sample. The device includes a detection chamber and a sample tray with centrifugal and positioning rotation functions, a detection unit, a drive device, and a heating device inside the detection chamber. The top of the detection chamber is equipped with a detection chamber cover, the sample tray is equipped with a detection tube rack, the detection tubes are placed inside the detection tube rack, the sample tray is connected to the drive device, the detection unit consists of a photoelectric detector and a light source, the detection unit is positioned corresponding to the detection tube rack, and the bottom of the detection chamber is equipped with a heating device.

[0005] Conventional methods for detecting platelet aggregation include optical methods, impedance methods, and filtration methods. However, these methods require expensive equipment and consume large amounts of blood samples (e.g., 50 ml of whole blood per person, or 10 ml of whole blood per mouse, or 10 mice), have poor reproducibility, and demand high levels of operator skill. Furthermore, platelets, being nucleus-free, cannot be cultured or proliferated in vitro, have a short lifespan after in vitro, and require stringent storage conditions, which significantly limits research on platelet activation. Currently, there is an urgent need to develop a novel platelet function detection technology that can both conserve platelet consumption and offer better reproducibility.

[0006] Microfluidics is a technology for precisely controlling and manipulating fluids at the microscale, particularly those at the submicron level. By constructing fluid channels and functional units at the micrometer scale, precise control and manipulation of fluids can be achieved. Microfluidics offers advantages such as small size, low material consumption, fast response speed, and high integration, and is widely used in fields such as biomedicine, chemical analysis, and environmental monitoring.

[0007] CN102348506A discloses a device for platelet aggregation using a microfluidic device, which assesses platelet aggregation in real time from a biological sample obtained from a subject. The device includes: a channel for the passage of the biological sample, the channel including a protrusion for inducing the formation of an upstream shear acceleration zone coupled to a downstream shear deceleration zone, and defining a shear rate peak zone between the upstream shear acceleration zone and the downstream shear deceleration zone, the downstream shear deceleration zone defining a platelet aggregation region; and a platelet detection device for detecting platelet aggregation occurring in the aggregation region due to the passage of the biological sample through the channel.

[0008] Although microfluidic chips have been applied to the detection of platelet aggregation, the following shortcomings still exist: 1) The design structure of microfluidic chips is relatively complex, and the preparation and operation are difficult; 2) The microchannels in the chip are easily adhered by platelets, resulting in signal interference; 3) There are limitations in terms of stability and reliability. Summary of the Invention

[0009] One object of the present invention is to provide a high-throughput chip for detecting platelet function, which significantly reduces the amount of whole blood required.

[0010] Another objective of this invention is to provide a high-throughput chip for platelets to react simultaneously with multiple molecules, thereby improving the efficiency of platelet activation research.

[0011] Another objective of this invention is to provide a high-throughput chip suitable for detecting the aggregation effect of platelets interacting with different molecules or different concentrations of the same molecule, which is beneficial for drug screening.

[0012] Another object of the present invention is to provide a medical device for high-throughput detection of platelets.

[0013] A high-throughput chip for detecting platelet function, comprising:

[0014] The base layer is used to provide support for the chip to implement detection functions;

[0015] The reaction layer, which is built on the base layer, includes several reaction chambers. Each reaction chamber includes a first cavity and a second cavity. The first cavity and the second cavity are connected, and the accommodating space of the first cavity is at least twice as large as the accommodating space of the second cavity, especially 2 to 6 times.

[0016] The flow layer, which is the space between the base layer and the reaction layer, is used to contain fluid and is connected to each reaction chamber;

[0017] A fluid distribution component, disposed at the outer edge of the flow layer, includes at least one distribution port, which introduces fluid into the flow layer and distributes the fluid evenly.

[0018] The liquid inlet is connected to the fluid distribution component and is used to input solution droplets and kinetic fluid into the fluid distribution component.

[0019] A specific embodiment of the first cavity of a high-throughput chip for detecting platelet function has a cubical accommodating space with a processing depth of 40 micrometers to 80 micrometers, an opening length of 65 micrometers to 100 micrometers, and a processing width of 55 micrometers to 80 micrometers.

[0020] Another specific embodiment of the first cavity of a high-throughput chip for detecting platelet function has a cubical accommodating space with a depth of 80 micrometers, an opening length of 90 micrometers, and a width of 70 micrometers.

[0021] A specific embodiment of the second cavity of a high-throughput chip for detecting platelet function, wherein the accommodating space is processed into a cube with a processing depth of 40 micrometers to 80 micrometers, an opening length of 25 micrometers to 40 micrometers, and a processing width of 25 micrometers to 40 micrometers.

[0022] Another specific embodiment of the second cavity of a high-throughput chip for detecting platelet function has a cubical accommodating space with a depth of 80 micrometers, an opening length of 40 micrometers, and a width of 40 micrometers.

[0023] The high-throughput chip for detecting platelet function of the present invention also includes a droplet filter disposed at the outer edge of the reaction layer to filter out droplets with excessively small particle size (e.g., less than 25 micrometers).

[0024] Another implementation of the droplet filter includes a groove with its opening connected to the flow layer, allowing droplets with excessively small diameters to enter the groove opening and be fixed within the groove cavity.

[0025] Another implementation of the droplet filter includes two or more channels arranged in parallel, with each channel opening communicating with the flow layer, so that droplets with excessively small particle sizes enter the channel opening and are fixed in the channel cavity.

[0026] Another embodiment of the droplet filter includes two or more channels arranged in parallel, with a spacing of 40 to 75 micrometers between channels, and the openings of the channels are all in communication with the flow layer. The channel openings are 15 to 35 micrometers, especially 25 micrometers.

[0027] The high-throughput chip for detecting platelet function of the present invention further includes at least one liquid collection outlet, which is in communication with the flow layer and aligned with the fluid distribution component. Negative pressure is applied at the liquid collection outlet to accelerate fluid flow.

[0028] The high-throughput chip for detecting platelet function of the present invention further includes several support members, each of which is connected at both ends to a reaction layer and a base layer, so that the reaction layer is supported by the base layer. Simultaneously, the support members also allow the reaction layer to be mounted on the base layer, so that the space formed between the base layer and the reaction layer becomes a flow layer to accommodate fluid.

[0029] The high-throughput chip for platelet detection of the present invention further includes a flow aid port connected to a fluid distribution unit for inputting motive fluid into the fluid distribution unit to assist the flow of droplets within the flow layer. The connection point between the flow aid port and the fluid distribution unit, as well as the connection point between the liquid guide port and the fluid distribution unit, are located at one point to facilitate the movement of solution droplets by the motive fluid and to prevent solution droplets from accumulating at the connection point.

[0030] This invention relates to a high-throughput chip for detecting platelet function, in which fluorinated oil is added to the inlet at a flow rate of 650±50 μl / h.

[0031] This invention relates to a high-throughput chip for detecting platelet function. Fluorinated oil is added to the inlet at a flow rate of 650±50 μl / h. Simultaneously, fluorinated oil is extracted from the collection outlet at a flow rate of 700±50 μl / h, thereby creating a pressure difference and shortening the fluid travel time in the flow layer.

[0032] This invention relates to a high-throughput chip for detecting platelet function, comprising at least one fluid distribution component. Correspondingly, it includes at least one liquid inlet, optionally at least one liquid collection outlet, and more preferably at least one flow-aiding inlet.

[0033] To fully utilize the reaction chambers and ensure that solution droplets can smoothly enter each reaction chamber and prevent "dead zones", at least two fluid distribution components are provided on the high-throughput chip for platelet detection in this invention. The fluid flow directions from the outlets of each fluid distribution component are staggered, for example, lateral flow.

[0034] Another high-throughput chip for detecting platelet function includes a first fluid distribution unit and a second fluid distribution unit. The flow direction (D1) from the first fluid distribution unit into the flow layer intersects with the flow direction (D2) from the second fluid distribution unit into the flow layer, and the angle formed is greater than or equal to 30 degrees and less than or equal to 120 degrees, especially 70 degrees to 90 degrees.

[0035] Another high-throughput chip for detecting platelet function includes:

[0036] The base layer is used to provide support for the chip to implement detection functions;

[0037] The reaction layer, which is convex quadrilateral, is built on the base layer and includes several reaction chambers. Each reaction chamber includes a first cavity and a second cavity. The first cavity and the second cavity are connected, and the accommodating space of the first cavity is at least twice as large as the accommodating space of the second cavity, especially 2 to 6 times.

[0038] The flow layer, which is the space between the base layer and the reaction layer, is used to contain fluid and is connected to each reaction chamber;

[0039] A first fluid distribution component is disposed at the first edge of the reaction layer and includes at least one first distribution port to introduce fluid into the flow layer and to make the fluid uniformly distributed.

[0040] The second fluid distribution component is disposed at the second edge of the reaction layer and includes at least one second distribution port to introduce fluid into the flow layer and make the fluid uniformly distributed.

[0041] One end of the first edge is connected to one end of the second edge;

[0042] The first liquid inlet is connected to the first fluid distribution component and is used to input solution droplets and dynamic fluid into the first fluid distribution component.

[0043] The second liquid inlet is connected to the second fluid distribution component and is used to input solution droplets and kinetic fluid into the second fluid distribution component.

[0044] Correspondingly, another high-throughput chip for platelet detection also includes at least one first flow aid port connected to a first fluid distribution component for inputting motive fluid into the first fluid distribution component. The connection point between the first flow aid port and the first fluid distribution component, as well as the connection point between the first liquid guide port and the first fluid distribution component, are located at the same point to facilitate the movement of solution droplets by the motive fluid and to prevent solution droplets from accumulating at the connection point.

[0045] Correspondingly, another high-throughput chip for platelet detection also includes at least one second flow aid port connected to a second fluid distribution unit for inputting motive fluid into the second fluid distribution unit. The connection point between the second flow aid port and the second fluid distribution unit, as well as the connection point between the second liquid guide port and the second fluid distribution unit, are located at the same point to facilitate the movement of solution droplets by the motive fluid and to prevent solution droplets from accumulating at the connection point.

[0046] Correspondingly, another high-throughput chip for detecting platelet function also has at least one first collection outlet, which is connected to the flow layer and aligned with the first fluid distribution component. Negative pressure is applied at the first collection outlet to accelerate the flow of fluid from the first fluid distribution component.

[0047] Correspondingly, another high-throughput chip for detecting platelet function also has at least one second collection outlet, which is connected to the flow layer and aligned with the second fluid distribution component. Negative pressure is applied at the second collection outlet to accelerate the flow of fluid from the second fluid distribution component.

[0048] In the various high-throughput chips for detecting platelet function provided by this invention, the shape of the reaction layer is preferably a parallelogram, especially a square or rectangle.

[0049] In the various high-throughput chips for detecting platelet function provided by the present invention, the fluid distribution component includes a main pipeline and several branch pipelines. The main pipeline is connected to the liquid inlet and the flow aid inlet, and distributes the received solution droplets and kinetic fluid to each branch pipeline, so as to realize that the solution droplets and kinetic fluid can spread and distribute rapidly, which is conducive to the solution droplets entering each reaction chamber.

[0050] Several branch pipes can also be set on one main pipe to further disperse the solution droplets and kinetic fluid entering the flow layer, and further accelerate the entry of solution droplets into each reaction chamber.

[0051] When applying the high-throughput chip of this invention, large-volume droplets (60-90 micrometers in diameter, also known as large droplets) are first added into the first cavity, followed by small-volume droplets (30-60 micrometers in diameter, also known as small droplets) which then enter the second cavity. For example, platelets obtained through separation are dissolved in the large droplets, while drugs are dissolved in the small droplets. Corona discharge is used to induce the fusion of the two types of droplets, allowing for the identification of platelets and the counting of free platelets, thereby assessing the drug's effect on platelet aggregation.

[0052] The high-throughput chip of this invention is used as a medical device to examine the platelet aggregation function of patients, providing a basis for personalized drug delivery.

[0053] The high-throughput chip of this invention enables the fusion of micro-volume droplets (below 100 micrometers), effectively controlling the position, size, and number of droplets, and achieving the fusion of various types of droplets. This allows for reactions to be carried out in micro-volumes, significantly reducing the volume requirements of the sample. It is particularly beneficial for the observation and analysis of platelet aggregation, providing a new technical solution for platelet activity research. Attached Figure Description

[0054] Figure 1 This is a schematic diagram of an embodiment of a high-throughput chip for detecting platelet function provided by the present invention.

[0055] Figure 2 This is a schematic diagram of another embodiment of the high-throughput chip for detecting platelet function provided by the present invention.

[0056] Figure 3 This is a schematic diagram of another embodiment of the high-throughput chip for detecting platelet function provided by the present invention.

[0057] Figure 4 This is a schematic diagram of another embodiment of the high-throughput chip for detecting platelet function provided by the present invention.

[0058] Figure 5 for Figure 1 A partial schematic diagram from another angle of a high-throughput chip used to detect platelet function;

[0059] Figure 6 This is a schematic diagram of the reaction chamber morphology of the high-throughput chip for detecting platelet function according to the present invention.

[0060] Figure 7 A bright-field microscope image of the high-throughput chip for detecting platelet function of the present invention with a filter dropper.

[0061] Figure 8 This is a bright-field microscope field of view of an embodiment of the high-throughput chip of the present invention for detecting platelet function. Detailed Implementation

[0062] The technical solution of the present invention will be described in detail below with reference to the accompanying drawings. The embodiments of the present invention are only used to illustrate the technical solution of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solution of the invention without departing from the spirit and scope of the technical solution of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention.

[0063] The high-throughput chip for detecting platelet function in this embodiment can be fabricated using currently common technologies. As an example, the method for fabricating the chip is described below:

[0064] First, spin-coat the photoresist, including:

[0065] 1) Placing the silicon wafer: Turn on the spin coater, lay out the aluminum foil, place the silicon wafer on the spin coater, and adjust its position to the center;

[0066] 2) Pouring photoresist: Pour SU8-3025 photoresist into the center of the silicon wafer. The volume of the photoresist should be about the size of a one-yuan coin (about 1 ml).

[0067] 3) Perform spin coating; the final coating speed depends on the design height.

[0068] 4) Preheating: Place the silicon wafer coated with photoresist on a 95°C heating plate and heat it for a period of time depending on the design height.

[0069] Then, ultraviolet lithography is performed, including:

[0070] 1) UV Exposure: Place the silicon wafer on an acrylic plate, place the mask on the silicon wafer, and secure the silicon wafer and mask with transparent adhesive tape along the edges. Press on a quartz glass plate. Expose the wafer by irradiating the center with a UV photomask, turning the switch knob clockwise to the end. The exposure time is determined by the depth: 1 minute of exposure time is required for every 10 µm of depth.

[0071] 2) Post-baking heating: Place the UV-exposed silicon wafer on a 95℃ heating plate and heat for 5-10 minutes;

[0072] 3) Development and soaking: Place the silicon wafer that has been baked and heated into the developing solution, turn the knob slightly counterclockwise to start the magnetic stirrer, and rinse and soak the silicon wafer for 10 min to 20 min.

[0073] 4) Cleaning and drying: Remove the silicon wafer from the developer using a wafer clamp, clean the silicon wafer in sequence with developer, isopropanol, and ethanol, and then dry it with a nitrogen gun;

[0074] 5) Packaging and labeling: Place the processed silicon wafers into petri dishes and label them.

[0075] Re-cover PDMS, including:

[0076] 1) PDMS preparation and degassing: Prepare PDMS and curing agent in a 10:1 ratio, stir well with a stirring rod, and then use a vacuum pump to remove air bubbles for 20-30 minutes to obtain the PDMS liquid to be gelled; (by mass, PDMS:curing agent is 10:1, and one cup of PDMS should not exceed 30g to prevent overflow during degassing)

[0077] 2) Overnight curing: Pour the PDMS solution into a petri dish containing the processed silicon wafer, blow away the air bubbles with an air gun; place in a 65℃ constant temperature incubator overnight;

[0078] 3) Cutting and punching (creating droplet inlets and kinetic fluid inlets): Use a scalpel to cut the PDMS around the patterned area to obtain the PDMS chip, with the patterned side facing up. Use a punch to make holes at the locations where droplets need to be introduced; remove the fragments with transparent tape, and then cover the chip with transparent tape.

[0079] 4) Plasma treatment: Remove the chip from the transparent adhesive and place it together with a clean glass plate into the Plasma machine for surface treatment, with the bonding side facing up. The air pressure is around 600-800, and the treatment lasts for 80 seconds.

[0080] 5) Heat bonding: Take out the PDMS chip and glass plate, put the two sides treated with Plasma together, and then place them on a 95℃ heating plate for 30 minutes to bond them together;

[0081] 6) Overnight oven drying: Place the heated chip in an oven and leave it overnight; this completes the chip processing.

[0082] If the groove is also needed as part of the chip, it is completed together with the main body of the chip (except for the inlet / outlet holes). During the ultraviolet exposure process, the various structures on the mask (the chip body, the groove, and the positions of each fluorinated oil and droplet inlet / outlet) are etched onto the silicon wafer. Then, the PDMS chip is fabricated using the silicon wafer as a template.

[0083] Figure 1 This is a schematic diagram of an embodiment of the high-throughput chip for detecting platelet function provided by the present invention. Figure 5 for Figure 1 The diagram shown is a partial view from another angle of a high-throughput chip used to detect platelet function. Figure 6 This is a schematic diagram of the reaction chamber morphology of the high-throughput chip for detecting platelet function according to the present invention. Figure 1 , Figure 5 and Figure 6As shown, the high-throughput chip for platelet detection in this embodiment includes a base layer 100, a reaction layer 300, and a flow layer 200. The base layer 100 provides support for the chip to perform its detection function, and the flow layer 200 is the space between the base layer 100 and the reaction layer 300. The reaction layer 300 is mounted on the base layer 100, and its shape is preferably a parallelogram, especially a square or rectangle.

[0084] The reaction layer 300 includes several reaction chambers 350, each including a first cavity 351 and a second cavity 352, which are connected. The flow layer 200 is a space located between the base layer 100 and the reaction layer 300, used to contain fluid, and is connected to each reaction chamber. Several support members 400 are located between the base layer 100 and the reaction layer 300, with both ends of each support member connected to the reaction layer 300 and the base layer, respectively, so that the reaction layer 300 is supported by the base layer 100, and the reaction chambers 350 are suspended above the flow layer 200 with their openings facing the flow layer 200, in an "inverted" shape, so that droplets in the flow layer 200 can enter the reaction chambers 350. In this embodiment, the reaction layer 300 is a square with a side length of 6500 micrometers, and has 60×34 reaction chambers 350 distributed therein. The accommodating space of the first cavity 351 is four times larger than the accommodating space of the second cavity 352. For example, the accommodating space of the first cavity 351 is machined into a cube with a machining depth of 80 micrometers, an opening length of 90 micrometers, and a machining width of 70 micrometers. The accommodating space of the second cavity 352 is machined into a cube with a machining depth of 80 micrometers, an opening length of 40 micrometers, and a machining width of 40 micrometers.

[0085] The fluid distribution component 500 includes at least one distribution port 513 to introduce fluid into the flow layer 200 and to uniformly distribute the fluid (e.g., droplets for reaction and flow aids). To facilitate droplet flow, a motive fluid, such as fluorinated oil, is added. The droplets float on the fluorinated oil, move with it, and enter the reaction chamber 350 accordingly. Larger droplets enter the larger first cavity 351, while smaller droplets enter the smaller second cavity 352. To ensure accurate entry of large and small droplets into their respective cavities, the larger droplets are added first, followed by the smaller droplets.

[0086] The liquid inlet 600 is configured to communicate with the fluid distribution component 500 for inputting large and small droplets into the fluid distribution component 500. A driving fluid, such as fluorinated oil, can also be input through the liquid inlet 600. For example, after a large droplet is input through the liquid inlet 600, fluorinated oil is then input to move the large droplet already input into the fluid distribution component 500, causing it to enter the larger first cavity 351. Next, a small droplet is input through the liquid inlet 600, and then fluorinated oil is input again to move the small droplet already input into the fluid distribution component 500, causing it to enter the smaller second cavity 352.

[0087] By configuring the flow aid 700 to be connected to the fluid distribution component 500, so that the motive fluid, such as fluorinated oil, is input into the fluid distribution component 500 through the flow aid 700, it can not only significantly improve the synchronization between large or small droplets and fluorinated oil, prevent pipeline blockage, and enhance the uniformity of droplet and motive fluid input into the fluid distribution component, but also significantly shorten the operation time.

[0088] The connection between the flow aid 700 and the fluid distribution component 500, and the connection between the liquid guide 600 and the fluid distribution component 500, are located at one place to facilitate the uniformity of the flow of solution droplets into the fluid distribution component driven by the power fluid, and to prevent solution droplets from accumulating at the connection.

[0089] A main pipe 511 and several branch pipes 512 are provided in the fluid distribution unit 500. The main pipe 511 is connected to the liquid guide port 600 and the flow aid port 700, distributing the received solution droplets and kinetic fluid to each branch pipe 512 to achieve rapid spreading and distribution of the solution droplets and kinetic fluid, facilitating the entry of the solution droplets into each reaction chamber. The fluid flowing out of the fluid distribution unit 500 flows within the flow layer 200. The liquid collection outlet 800 is aligned with the fluid distribution unit 500, communicating with the flow layer 200 in the forward flow direction, and attracting fluid with negative pressure at the liquid collection outlet to further accelerate the fluid flow. For example, by using a syringe pump or pressure-driven technology, fluorinated oil is added to the flow port at a flow rate of 650 μl / h. Then, the fluorinated oil is extracted at a higher flow rate at the liquid collection outlet 800, such as 700 μl / h. This creates a pressure difference, shortens the movement time of the fluid in the flow layer 200, and allows the droplets to be distributed more quickly in the accommodating space of the first cavity 351 or the accommodating space of the second cavity 352.

[0090] In this embodiment, both large and small droplets can be determined using known methods, such as those described in Appl. Phys. Lett. 2003, 82, 364-366 and Phys. Rev. Lett. 2003, 90, 144505-1-4. However, some tiny droplets may still be mixed in. To ensure that all droplets entering the reaction chamber are of the required size, a droplet filter is provided at the outer edge of the reaction layer 300 to remove droplets with excessively small particle sizes.

[0091] Figure 2 This is a schematic diagram of another embodiment of the high-throughput chip for detecting platelet function provided by the present invention. Figure 2 As shown, the droplet filter 900 is disposed on the outer edge of the reaction layer 300 and includes several grooves 931. The groove openings are connected to the flow layer (not shown), so that droplets with excessively small particle size in the flow enter the groove openings and are fixed in the groove cavity. Figure 7 As shown, tiny droplets Dp3 (i.e., smaller than small droplets) are captured by the groove 931 (groove opening of 25 micrometers) of the droplet filter 900 when they flow out from the fluid distribution unit 500 into the flow layer 200, and do not enter the reaction chamber 350. Similarly, larger droplets (i.e., large droplets Dp1 and small droplets Dp2) are not captured by the droplet filter 900 and smoothly enter the flow layer 200 and are distributed in the reaction chamber 350.

[0092] Figure 3 This is a schematic diagram of another embodiment of the high-throughput chip for detecting platelet function provided by the present invention. See also... Figure 5 ,like Figure 3 As shown, two fluid distribution components are disposed thereon: a first fluid distribution component 510 and a second fluid distribution component 520. The first fluid distribution component 510 is disposed at the first edge 310 of the reaction layer 300 and includes at least one first distribution port (not shown), which guides fluid (e.g., droplets and kinetic fluid) into the flow layer and ensures uniform fluid distribution. The second fluid distribution component is disposed at the second edge 320 of the reaction layer and includes at least one second distribution port (not shown), which guides fluid (e.g., droplets and kinetic fluid) into the flow layer and ensures uniform fluid distribution. In this embodiment, the first edge 310 and the second edge 320 are adjacent edges of the square reaction layer 300, connected at one end.

[0093] Similarly, the first liquid inlet 610 is configured to connect with the first fluid distribution member 510 to input large and small droplets into the first fluid distribution member 510, and the second liquid inlet 620 is configured to connect with the second fluid distribution member 520 to input large and small droplets into the second fluid distribution member 520. The flow direction from the first fluid distribution member 510 into the flow layer (denoted as D1) intersects with the flow direction from the second fluid distribution member 520 into the flow layer (denoted as D2), and the fluid flow directions at the outlets of each fluid distribution member are staggered, for example, with an angle of 90 degrees, i.e., crossflow. This prevents the high-throughput chip in this embodiment from having "dead zones" in droplet distribution, i.e., some reaction chambers without droplets, so as to make full use of the reaction chambers and allow solution droplets to smoothly enter each reaction chamber.

[0094] The first flow aid port 710 is configured to connect with the first fluid distribution component 510, allowing the driving fluid, such as fluorinated oil, to enter the first fluid distribution component 510 through the first flow aid port 710. The second flow aid port 720 is configured to connect with the second fluid distribution component 520, allowing the driving fluid, such as fluorinated oil, to enter the second fluid distribution component 520 through the second flow aid port 720. This not only significantly improves the synchronization between large or small droplets and fluorinated oil, preventing pipeline blockage and enhancing the uniformity of droplet and driving fluid input to the fluid distribution component, but also significantly shortens the operation time.

[0095] The connection between the first flow aid port 710 and the first fluid distribution member 510, and the connection between the first liquid guide port 610 and the first fluid distribution member 510 are located at one place. The connection between the second flow aid port 720 and the second fluid distribution member 520, and the connection between the second liquid guide port 620 and the second fluid distribution member 520, are located at one place, either individually or simultaneously, to facilitate the uniformity of the flow of solution droplets into the fluid distribution member driven by the power fluid, and to prevent solution droplets from accumulating at the connection points.

[0096] The fluid flowing out of the first fluid distribution member 510 flows within the flow layer. The first liquid collection outlet 810 is aligned with the first fluid distribution member 510 and communicates with the flow layer 200 in the forward flow direction. Pressurization is applied at the first liquid collection outlet 810 to further accelerate the fluid flow. For example, if fluorinated oil is added to the flow aid port at a flow rate of 650 μl / h, the fluorinated oil will be extracted from the first liquid collection outlet 810 at a higher flow rate, such as 700 μl / h. This creates a pressure difference, shortening the fluid travel time in the flow layer 200, allowing droplets to distribute more quickly within the accommodating space of the first cavity 351 or the accommodating space of the second cavity 352. Similarly, the fluid flowing out of the second fluid distribution member 520 flows within the flow layer. The second liquid collection outlet 820 is aligned with the second fluid distribution member 520 and communicates with the flow layer 200 in the forward flow direction. Pressurization is applied at the second liquid collection outlet 820 to further accelerate the fluid flow.

[0097] Figure 4 This is a schematic diagram of another embodiment of the high-throughput chip for detecting platelet function provided by the present invention. See also... Figure 7 ,like Figure 4 As shown, the first dropper 910 is disposed on the outer edge of the first edge 310 and includes several grooves 911. The groove openings communicate with the flow layer (not shown), allowing droplets with excessively small particle sizes to enter the groove openings and be fixed within the groove cavity. The second dropper 920 is disposed on the outer edge of the second edge 320 and includes several grooves 921. Tiny droplets, when flowing out from the fluid distribution unit and entering the flow layer, are captured by the grooves of the dropper (groove openings are 25 micrometers) and do not enter the reaction chamber. Similarly, larger droplets (i.e., large and small droplets) are not captured by the dropper and smoothly enter the flow layer and are distributed within the reaction chamber.

[0098] After large droplets containing platelets and small droplets carrying drug molecules (anticoagulants such as aspirin) are successfully distributed in reaction chamber 350, the high-throughput chip of this embodiment is corona-treated using an external corona generator. This causes the corresponding large and small droplets within reaction chamber 350 to fuse, ultimately resulting in platelet aggregation. The aggregation of platelets in the fused droplets within the reaction chamber is observed under a microscope. Figure 8 As shown, visual observation revealed that the platelet aggregation level in sample droplet T1 was significantly higher than that in sample droplet T2. Free platelets were identified and counted using BaxterAlgorithms software to assess platelet aggregation function.

[0099] The high-throughput chip used in this embodiment can not only detect the aggregation effect of platelets interacting with different concentrations of the same molecule, understand the platelet aggregation of patients under different drug doses, and provide personalized drug dosing plans, but also simultaneously detect the aggregation effect of platelets interacting with different drug molecules, providing patients with personalized drug dosing plans, and can obtain test results quickly, greatly improving convenience.

Claims

1. Use of a chip in the manufacture of a medical device for detecting the function of platelets, for observing and analyzing the aggregation of platelets, characterized in that, The chip comprises: a base layer for providing support for the chip to realize detection function; a reaction layer erected on the base layer, comprising a plurality of reaction cavities, each reaction cavity comprising a first cavity and a second cavity, the first cavity and the second cavity being in communication, and the accommodation space of the first cavity being at least 2 times larger than that of the second cavity; a flow layer, which is a space between the base layer and the reaction layer, for accommodating fluid and being in communication with each reaction cavity; a fluid distribution member provided at the outer edge of the flow layer, comprising at least one distribution port for guiding fluid into the flow layer and making the fluid uniformly distributed; a liquid guide port in communication with the fluid distribution member for inputting solution droplets and motive fluid into the fluid distribution member; a flow assisting port in communication with the fluid distribution member for inputting motive fluid into the fluid distribution member, the communication part of the flow assisting port with the fluid distribution member and the communication part of the liquid guide port with the fluid distribution member being at one place to facilitate the motive fluid to move the solution droplets; a droplet filter provided at the outer edge of the reaction layer, comprising at least one groove, the groove opening being in communication with the flow layer so that the tiny droplets in the flow enter the groove opening and are fixed in the groove cavity; further comprising a flow assisting port in communication with the fluid distribution member; the number of the fluid distribution members is 2, i.e. a first fluid distribution member and a second fluid distribution member, the flow direction of the fluid entering the flow layer from the first fluid distribution member intersecting with the flow direction of the fluid entering the flow layer from the second fluid distribution member, the intersection angle being greater than or equal to 30 degrees and less than or equal to 120 degrees; the first fluid distribution member is provided at the first edge of the reaction layer, comprising at least one first distribution port for guiding fluid into the flow layer and making the fluid uniformly distributed; the second fluid distribution member is provided at the second edge of the reaction layer, comprising at least one second distribution port for guiding fluid into the flow layer and making the fluid uniformly distributed; one end of the first edge is connected with one end of the second edge; further comprising: a first liquid guide port in communication with the first fluid distribution member for inputting solution droplets and motive fluid into the first fluid distribution member; a second liquid guide port in communication with the second fluid distribution member for inputting solution droplets and motive fluid into the second fluid distribution member; further provided with: at least one first flow assisting port in communication with the first fluid distribution member for inputting motive fluid into the first fluid distribution member; at least one second flow assisting port in communication with the second fluid distribution member for inputting motive fluid into the second fluid distribution member; at least one first liquid collecting outlet in communication with the flow layer and arranged opposite to the first fluid distribution member; at least one second liquid collecting outlet in communication with the flow layer and arranged opposite to the second fluid distribution member.

2. Use according to claim 1, characterized in that further comprising at least one liquid collecting outlet in communication with the flow layer and arranged opposite to the fluid distribution member.

3. Use according to claim 1, characterized in that Also included are a plurality of support members, each of said support members having two ends, one end of each of said support members being connected to said reaction layer and the other end of each of said support members being connected to said base layer, such that said reaction layer is supported by said base layer, and such that said reaction layer is suspended over said base layer, such that a space is formed between said base layer and said reaction layer for containing fluid in said flow layer.

4. A medical device for detecting platelet function, characterized in that A chip as claimed in claim 1.

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