Circulating microflow blood thrombogenesis observation device

Abstract

The utility model relates to a circulating microflow blood thrombogenesis observation device. The device comprises a fixing table and a thrombus generation assembly, wherein the thrombus generation assembly is arranged on the fixing table; the thrombus generating assembly comprises a fluid circulating system and a fluid shunting device which are communicated with each other; the fluid circulating system comprises a sample adding unit, a circulating pipeline and a fluid driving device which are in fluid communication; the incubation unit is arranged on the sample adding unit and part of the circulating pipeline; the fluid flow dividing device comprises a flow dividing plate, a second flow dividing pipe and a plurality of first flow dividing pipes are arranged in the flow dividing plate in a penetrating mode, the first flow dividing pipes are communicated with the second flow dividing pipe, the second flow dividing pipe is communicated with the circulating pipeline, and flow stoppers are arranged on the second flow dividing pipe and the first flow dividing pipes.

Description

Technical Field

[0001] This utility model relates to the fields of biomedical engineering and experimental medicine, and in particular to a device for observing the formation of blood clots in circulating microflow. Background Technology

[0002] Hemostasis is a vital physiological function primarily driven by platelets. Abnormalities in blood clotting can lead to serious illnesses and even life-threatening situations. The coagulation process is extremely complex, with thrombus formation in the blood flow phase being crucial. Therefore, observing the process of thrombus formation in the blood is of great significance.

[0003] Under normal physiological conditions, vascular endothelial cells produce anticoagulants such as prostacyclin and nitric oxide. When the vascular endothelium is damaged, endothelial collagen is exposed, and von Willebrand factor (VWF) is transferred from the intracellular space of the damaged endothelial cells to the cell membrane surface. At the same time, CD62P (P-selectin) from the intracellular space of the endothelial cells is transferred to the cell membrane, inducing platelets activated by multiple pathways to bind to VWF / collagen. This leads to a coagulation reaction at the site of injury and the formation of a thrombus. The thrombus then contracts until it blocks the wound.

[0004] Currently, observing the in vitro thrombosis process mostly requires anesthetizing and dissecting experimental animals to create a vascular injury model for observation and recording. This method demands highly skilled experimental techniques from the operator, and the experimental animals also face the risk of open wounds, infection, and death. Existing thrombosis observation devices have significant drawbacks, including high costs hindering widespread adoption, complex operation requiring specialized personnel for operation and maintenance, long detection times impacting rapid diagnosis and treatment, limited sensitivity and specificity leading to potential false positives or false negatives, and large device size hindering portability and storage. These issues significantly limit the widespread application of thrombosis observation devices in clinical practice and research. Utility Model Content

[0005] In view of the shortcomings of the prior art described above, the purpose of this utility model is to provide a circulating microfluidic blood thrombosis observation device to solve the problems of existing thrombosis observation devices, such as difficulty in maintaining constant physiological temperature and conditions, inability to accurately control the flow rate and volume of drugs and blood, susceptibility to cross-infection, and difficulty in accurately simulating the complex process of vascular damage and thrombosis under different conditions.

[0006] To achieve the above and other related objectives, this utility model provides a device for observing the formation of thrombi in circulating microflow blood.

[0007] This utility model provides a circulating microfluidic blood thrombus formation observation device. The device includes a fixed platform and a thrombus formation component, which is disposed on the fixed platform. The thrombus formation component includes a fluid circulation system and a fluid diversion device connected in series. The fluid circulation system includes a fluidly connected sample application unit, a circulation pipeline, and a fluid driving device. It also includes an incubation unit disposed on the sample application unit and part of the circulation pipeline. The fluid diversion device includes a diversion plate, through which a second diversion tube and a plurality of first diversion tubes pass. Each first diversion tube is connected to a second diversion tube, and the second diversion tube is connected to the circulation pipeline. A flow stopper is provided on the second diversion tube and each of the first diversion tubes.

[0008] In some embodiments of this utility model, a battery assembly is also provided on the fixed platform, and the incubation unit and the fluid drive device are respectively connected to the battery assembly.

[0009] In some embodiments of this utility model, the fluid driving device is a peristaltic pump.

[0010] In some embodiments of this utility model, the incubation unit is a constant temperature incubation plate.

[0011] In some embodiments of this utility model, the diverter plate is provided with multiple receiving channels adapted to the second diverter pipe and multiple first diverter pipes.

[0012] In some embodiments of this utility model, the diverter plate is further provided with multiple through holes adapted to the second diverter pipe and multiple first diverter pipes for accommodating the flow stopper.

[0013] In some embodiments of this utility model, the flow stopper includes a flow stop knob and a base, the base being connected to one end of the through hole; the flow stop knob is provided with a screw with a hole, and the flow stop knob is threadedly connected to the base from the other end of the through hole through the screw.

[0014] In some embodiments of this utility model, the diameter of the circulation pipe is 2-4 mm.

[0015] In some embodiments of this utility model, the diversion plate is a transparent glass plate.

[0016] In some embodiments of this utility model, the length of the diverter plate is 70-80mm; the width of the diverter plate is 20-30mm.

[0017] In some embodiments of this utility model, each of the first diversion tubes is a capillary tube.

[0018] In some embodiments of this utility model, there are at least three first shunt tubes.

[0019] In some embodiments of this utility model, the diameter of the first shunt tube is smaller than the diameter of the second shunt tube.

[0020] In some embodiments of this utility model, the diameter of each first shunt tube is 0.5-1.5 mm.

[0021] In some embodiments of this utility model, the diameter of the second diverter is 3-5 mm.

[0022] Compared with the prior art, the circulating microfluidic blood thrombus formation observation device of this invention has the following beneficial effects:

[0023] This application, through the design of a fluid circulation system and a fluid diversion device, can precisely control the flow rate and volume of drugs and blood, avoid cross-infection, and simulate the complex process of vascular damage and thrombosis in vivo under different conditions; through components such as the sample dispensing unit, fluid driving device, and flow stopper, it can achieve automated operation and reduce human error; the device is compact in design, occupies little space, and is easy to carry and store. Attached Figure Description

[0024] Figure 1 The diagram shown is a schematic diagram of the overall structure of Embodiment 1 of this utility model.

[0025] Figure 2 The image shown is a top view of Embodiment 1 of this utility model.

[0026] Figure 3 The diagram shown is a partial schematic of the flow stopper of Embodiment 1 of this utility model.

[0027] Component designation explanation:

[0028] 1. Fixed platform

[0029] 2. Fluid circulation system

[0030] 21 Sampling Unit

[0031] 211 Incubation Unit

[0032] 22. Circulation Pipeline

[0033] 23 Fluid Driven Device

[0034] 3. Fluid splitting device

[0035] 31 splitter

[0036] 311 First shunt tube

[0037] 312 Second shunt tube

[0038] 32. Flow stopper

[0039] 321 Stop valve knob

[0040] 3211 Screw

[0041] 322 through hole

[0042] 4 Battery Components Detailed Implementation

[0043] The following details the implementation of the disclosed circulating microfluidic blood thrombus formation observation device.

[0044] Please see Figures 1-3 It should be understood that the structures, proportions, sizes, etc., illustrated in the accompanying drawings are merely for illustrative purposes to aid those skilled in the art and are not intended to limit the scope of this invention. Therefore, they have no substantial technical significance. Any modifications to the structure, changes in proportions, or adjustments to size, without affecting the effectiveness and purpose of this invention, should still fall within the scope of the disclosed technical content. Furthermore, the terms "upper," "lower," "left," "right," "middle," and "one" used in this specification are merely for clarity and not intended to limit the scope of this invention. Changes or adjustments to their relative relationships, without substantially altering the technical content, should also be considered within the scope of this invention.

[0045] like Figure 1As shown, this utility model embodiment provides a circulating microfluidic blood thrombus formation observation device. The device includes a fixed platform 1 and a thrombus formation component, which is disposed on the fixed platform 1. The thrombus formation component includes a fluid circulation system 2 and a fluid diversion device 3 connected in communication. The fluid circulation system 2 includes a sample application unit 21, a circulation pipeline 22, and a fluid driving device 23 connected in fluid communication. It also includes an incubation unit 211, which is disposed on the sample application unit 21 and part of the circulation pipeline 22. The fluid diversion device 3 includes a diversion plate 31, through which a second diversion tube 312 and a plurality of first diversion tubes 311 pass. Each first diversion tube 311 is connected to a second diversion tube 312. The second diversion tube 312 is connected to the circulation pipeline 22. A flow stopper 32 is provided on the second diversion tube 312 and each of the first diversion tubes 311. Typically, the incubation unit 211 can be located on the sample dispensing unit 21 and a portion of the circulation pipe 22 near the sample dispensing unit 21. In one specific embodiment, a pair of flow stoppers 32 is provided on the first diversion pipe 311 and each of the second diversion pipes 312. Two of the flow stoppers 32 in the pair are respectively located at both ends of the diversion plate 31. The flow stoppers 32 are used to control the opening and closing of the corresponding first diversion pipe 311 or second diversion pipe 312.

[0046] like Figure 2 As shown in the embodiment of the present invention, in the circulating microfluidic blood thrombosis observation device, the fixed platform 1 is also provided with a battery assembly 4, and the incubation unit 211 and the fluid drive device 23 are respectively electrically connected to the battery assembly 4.

[0047] In the circulating microfluidic blood thrombosis observation device provided in this embodiment of the invention, the fluid driving device 23 is a peristaltic pump, preferably a micro peristaltic pump. In a specific embodiment of the invention, the fluid driving device 23 is an adjustable-speed micro peristaltic pump, sourced from Kamoe Fluid Technology (Shanghai) Co., Ltd., model M1-NKP-DC, with a power of 10-14W. The peristaltic pump ensures unidirectional flow of blood or other liquids within the system, simulating the blood flow environment in the human body. By adjusting the power of the peristaltic pump, the flow rate can be precisely controlled to simulate different physiological conditions in experiments. The peristaltic pump provides continuous and stable flow, avoiding flow rate fluctuations that may occur in traditional methods, ensuring the reliability and repeatability of experimental results. Furthermore, since the liquid flows through the pump tube within the peristaltic pump, direct contact between the liquid and the pump body is avoided, reducing the risk of cross-contamination.

[0048] In the circulating microfluidic blood thrombosis observation device provided in this embodiment of the present invention, the incubation unit 211 is a constant temperature incubation plate. In a specific embodiment of the present invention, the constant temperature incubation plate is a PTC heating element with a digital temperature controller switch, purchased from Jinhua Longkexin Technology Co., Ltd., model XH-W3001 DC12V. The constant temperature incubation plate is used to heat the sample application unit 21 and the circulation pipeline 22 to 36.1-37.2℃, optionally 36.1-37℃ or 37-37.2℃, preferably 37℃, to simulate the in vivo environment, ensuring that the blood sample is maintained at the physiological temperature of the human body, effectively avoiding degradation or loss of activity of the blood sample due to temperature instability, thereby improving the reliability and repeatability of the experiment.

[0049] In a specific embodiment of this utility model, the sample application unit 21 includes a communicating sample application port and a sample application chamber. The sample application port can be opened and closed, and the sample application chamber is filled with physiological saline to simulate the real blood environment, maintain normal cell activity, dilute active substances in the blood, and play an anticoagulant role.

[0050] like Figure 3 As shown in the embodiment of the present invention, in the circulating microflow blood thrombosis observation device, the two ends of the diversion plate 31 are also provided with multiple through holes 322 adapted to the second diversion tube 312 and multiple first diversion tubes 311, for accommodating the flow stopper 32.

[0051] See also Figure 3 In the circulating microflow blood thrombosis observation device provided in this embodiment of the utility model, the flow stopper 32 includes a flow stop knob 321 and a base, the base being connected to one end of the through hole 322; the flow stop knob 321 is provided with a screw 3211 with a hole, and the flow stop knob 321 is threadedly connected to the base from the other end of the through hole 322 through the screw 3211. In a specific embodiment of the utility model, the side of the screw 3211 has a hole, and by rotating each of the flow stop knobs 321, the opening and closing and flow rate of each of the first diversion tubes 311 and the second diversion tubes 312 can be controlled respectively.

[0052] See also Figure 1 In the circulating microfluidic blood thrombus formation observation device provided in this embodiment of the invention, the diversion plate 31 is a transparent glass plate, and the diversion plate 31 has multiple receiving channels adapted to the second diversion tube 312 and multiple first diversion tubes 311. In a specific embodiment of this invention, the thrombus formation in each of the first diversion tubes 311 in the diversion plate can be observed using a microscope. For example, the time of thrombus formation, thrombus size and morphology, stability, and the influence of blood flow velocity and shear force on thrombus formation can be observed by simulating blood flow environments under different conditions.

[0053] In the circulating microflow blood thrombosis observation device provided in this embodiment of the utility model, the length of the diversion plate 31 is 70-80mm, optionally 70-75mm or 75-80mm, preferably 75mm; the width is 20-30mm, optionally 20-25mm or 25-30mm, preferably 25mm. The size of the diversion plate 31 is designed according to the size of the microscope slide.

[0054] In the circulating microfluidic blood thrombosis observation device provided in this embodiment of the utility model, the diameter of the circulating pipe 22 is 2-4 mm, which can be selected as 2-3 mm or 3-4 mm, preferably 3 mm. In a specific embodiment of this application, the diameter of the circulating pipe 22 is smaller than that of the second diversion pipe 312, and the two ends of the circulating pipe 22 are respectively accommodated in the two ends of the second diversion pipe 312.

[0055] In the circulating microfluidic blood thrombosis observation device provided in the embodiments of this utility model, there are at least three first shunt tubes 311, and each first shunt tube 311 is a capillary tube. In a specific embodiment of this utility model, there are three first shunt tubes 311, wherein two capillary tubes are coated with a layer of von Willebrand factor (VWF) and collagen in the central region, and the third capillary tube contains an agonist (ADP / EPI / AA).

[0056] Von Willebrand factor (VWF) is a protein in the blood that plays an important role in blood clotting. VWF helps platelets adhere to the site of blood vessel damage, thereby aiding in hemostasis.

[0057] The agonists are chemical substances that bind to specific receptors and activate them. These include:

[0058] ADP (adenosine diphosphate): ADP is a potent agonist that activates platelets by binding to P2Y1 and P2Y12 receptors, promoting platelet aggregation.

[0059] EPI (adrenaline): EPI is an agonist that enhances platelet response by binding to α2-adrenergic receptors.

[0060] AA (arachidonic acid): AA is an agonist that activates platelets by promoting the production of thromboxane A2 (TXA2) via the cyclooxygenase pathway.

[0061] The coating method can be, for example, dip coating or spray coating.

[0062] In the circulating microfluidic blood thrombosis observation device provided in this embodiment of the invention, the diameter of each of the first shunt tubes 311 is smaller than the diameter of the second shunt tubes 312. The diameter of the first shunt tube 311 is 0.5-1.5 mm, optionally 0.5-1 mm or 1-1.5 mm, preferably 1 mm. The diameter of the second shunt tube 312 is 3-5 mm, optionally 3-4 mm or 4-5 mm, preferably 4 mm.

[0063] like Figure 1-3 As shown, the usage process of the circulating microfluidic blood thrombus formation observation device according to this embodiment of the present invention includes the following steps:

[0064] 1) Start the incubation unit 211 and the liquid driving device 23, inject the blood sample from the sample application unit 21, and mix it evenly with the physiological saline in the sample application unit 21 under the heating action of the incubation unit 211. Then, place the circulating microflow blood thrombus formation observation device on the microscope stage and adjust the flow divider 31 to the center of the microscope's field of view.

[0065] 2) Open the stoppers 32 at both ends of the first diversion pipe 311 containing the agonist to allow the agonist to mix into the circulation pipe 22, and adjust the flow rate through the fluid drive device 23;

[0066] 3) Open the stoppers 32 at both ends of the first shunt tube 311 coated with von Willebrand factor and collagen, and the thrombus formation process can be observed and recorded under a microscope.

[0067] The principle of thrombus formation in this invention is as follows: The first shunt tube is coated with von Willebrand factor (VWF) and collagen. These two substances are exposed and induce platelet attachment and aggregation when vascular endothelial cells are damaged. When a blood sample flows through the first shunt tube 311, platelets bind to VWF and collagen, mimicking the coagulation process after vascular injury in the body. Agonists (such as ADP / EPI / AA) are introduced into the circulation channel 22 by controlling the opening and closing of the stopcock 32, further activating and accelerating the thrombus formation reaction.

[0068] In a specific embodiment of this utility model, the blood sample volume added in step 1) is 500-2500 μL, which can be selected as 200-1000 μL or 500-1500 μL.

[0069] In a specific embodiment of this utility model, the mixing time in step 1) is 3-7 minutes, which can be 3-5 minutes or 4-7 minutes, and preferably 5 minutes.

[0070] In a specific embodiment of this utility model, the heating to a temperature of 36.1-37.2℃ in step 1) can be selected as 36.1-37℃ or 37-37.2℃, preferably 37℃;

[0071] In a specific embodiment of this utility model, step 2) involves adjusting the flow rate to 2-5 mL / min. An adjustable peristaltic pump is used, and the required flow rate can be comprehensively evaluated based on the sample volume and microscopic observation, so that the entire circulation flow can be completed in about 1 minute.

[0072] In a specific embodiment of this invention, the agonist in step 2) includes one or more of adenosine diphosphate (ADP), epinephrine (EPI), and arachidonic acid (AA). The concentration of the adenosine diphosphate (ADP) agonist is 36 μmol / L. After the flow stopper is opened, the agonist is released into the circulation system to activate platelets.

[0073] In a specific embodiment of this invention, the coated von Willebrand factor and collagen are coated in the middle region of the first shunt tube, located in the center of the shunt plate's field of view, which facilitates microscopic observation. The concentration of the coated von Willebrand factor and collagen is 20 μg / mL.

[0074] In a specific embodiment of this utility model, the mass ratio of von Willebrand factor and collagen in step 3) is 1:1.

[0075] To make the technical means, creative features, objectives and effects of this utility model easy to understand, the following describes this utility model in detail with reference to the embodiments and accompanying drawings.

[0076] In the following examples, unless otherwise stated, all reactants are commercially available products.

[0077] Unless otherwise specified, the purity of each product in each embodiment of this utility model exceeds 98%.

[0078] Example 1

[0079] like Figure 1As shown, the detection device mainly consists of a constant-temperature incubation plate, a circulation pipeline, a sample application unit, a flow divider, and a micro-peristaltic pump. The constant-temperature incubation plate heats the sample application unit and its proximal tubing to a constant temperature of 37°C. The sample application chamber of the sample application unit is directly connected to the circulation pipeline, containing 200 μL of physiological saline. The flow divider is 75 mm long and 25 mm wide, which is the standard size of a glass slide. The flow divider contains three first flow dividers and one second flow divider. All three first flow dividers are capillary tubes; two capillary tubes are coated with VWF and Collagen in the central region, and the third capillary tube contains the agonist ADP / EPI / AA. The three first flow dividers are connected to the second flow divider, which is directly connected to the circulation pipeline. Two sets of symmetrically distributed flow stoppers are located at both ends of the flow divider to control the opening and closing of the first and second flow dividers. After the micro peristaltic pump is turned on, the liquid in the system flows sequentially through the sample dispensing unit, the micro peristaltic pump, and the diversion tube in the diversion plate, and finally flows back to the sample dispensing unit, realizing continuous flow and circulation of the liquid in the system. At the same time, the flow rate can be adjusted by adjusting the power of the micro peristaltic pump.

[0080] When the experimenter needs to observe thrombosis, first turn on the constant temperature incubation plate and the micro-peristaltic pump. Inject the blood sample containing the anticoagulant to be tested into the sample application unit through the soft rubber stopper. After the blood sample and the physiological saline in the sample application unit are completely mixed (mix for 3 minutes, and heat the sample to a constant temperature of 37°C), place the detection device on the microscope stage and adjust the shunt plate to the center of the microscope's field of view. After adjustment, open the stopcocks at both ends of the first shunt containing the agonist. The agonist mixes into the circulation pipeline. After adjusting the power of the micro-peristaltic pump to a suitable flow rate, open the stopcocks at both ends of the first shunt coated with von Willebrand factor and collagen. The observation and recording of the thrombosis process can then begin.

[0081] In summary, this utility model effectively overcomes the various shortcomings of the prior art and has high industrial application value.

[0082] The above embodiments are merely illustrative of the principles and effects of this utility model and are not intended to limit the scope of this utility model. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of this utility model. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in this utility model should still be covered by the claims of this utility model.

Claims

1. A device for observing the formation of blood clots in a circulating microfluidic system, characterized in that, The device includes a fixed platform (1) and a thrombus generation component, the thrombus generation component being disposed on the fixed platform (1); the thrombus generation component includes a fluid circulation system (2) and a fluid diversion device (3) connected in series; the fluid circulation system (2) includes a fluidly connected sample application unit (21), a circulation pipeline (22) and a fluid driving device (23); it also includes an incubation unit (211), the incubation unit (211) being disposed on the sample application unit (21) and part of the circulation pipeline (22); the fluid diversion device (3) includes a diversion plate (31), a second diversion tube (312) and a plurality of first diversion tubes (311) passing through the diversion plate (31), each of the first diversion tubes (311) being connected to the second diversion tube (312), the second diversion tube (312) being connected to the circulation pipeline (22), and a flow stopper (32) being provided on the second diversion tube (312) and each of the first diversion tubes (311).

2. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, The fixed platform (1) is also provided with a battery assembly (4), and the incubation unit (211) and the fluid drive device (23) are respectively connected to the battery assembly (4).

3. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, The fluid drive device (23) is a peristaltic pump; and / or, the incubation unit (211) is a constant temperature incubation plate.

4. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, The diverter plate (31) is provided with multiple receiving channels adapted to the second diverter pipe (312) and multiple first diverter pipes (311).

5. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, The diverter plate (31) is also provided with a plurality of through holes (322) adapted to the second diverter pipe (312) and a plurality of first diverter pipes (311) for accommodating the flow stopper (32).

6. The circulating microfluidic blood thrombus formation observation device as described in claim 5, characterized in that, The flow stopper (32) includes a flow stop knob (321) and a base, the base being connected to one end of the through hole (322); the flow stop knob (321) is provided with a screw (3211) with a hole, and the flow stop knob (321) is threadedly connected to the base from the other end of the through hole (322) through the screw (3211).

7. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, The diameter of the circulation pipe (22) is 2-4 mm; And / or, the diversion plate (31) is a transparent glass plate; And / or, the length of the diverter plate (31) is 70-80 mm; And / or, the width of the diverter plate (31) is 20-30 mm.

8. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, Each of the first shunt tubes (311) is a capillary tube; And / or, there are at least three first shunt tubes (311).

9. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, The diameter of each of the first shunt pipes (311) is smaller than the diameter of the second shunt pipe (312).

10. The circulating microfluidic blood thrombus formation observation device as described in claim 1, characterized in that, The diameter of each of the first diverter tubes (311) is 0.5-1.5 mm, and the diameter of the second diverter tube (312) is 3-5 mm.

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