Self-driven disc type blood separation and detection device and method

Abstract

The application discloses a self-driven disc type blood separation and detection device and method, which comprises a sample adding layer, a separation layer and a detection layer, and the layers are sequentially connected through screw fixation. The sample adding layer is provided with a sample adding port and a first screw hole; the separation layer comprises four repeated blood separation units and a second screw hole, and each separation unit comprises a sample inlet, a micro-fluidic channel, a filter groove, a sample outlet and a waste liquid collecting chamber; and the detection layer is provided with a screen-printed electrode interface and a third screw hole. Blood samples are dripped into the device from the sample adding port, and under the driving of the internal negative pressure of the device, the blood samples enter the micro-fluidic channel, and blood filtration is realized by using the gravity difference between blood cells and blood plasma. The filtered blood plasma flows into the detection layer and contacts with the screen-printed electrode detection unit, so that electrochemical detection is realized. The device does not need external power supply, thereby reducing sample operation, reducing the volume of the separation device, improving the blood separation efficiency, and facilitating instant detection and multi-channel detection of blood samples.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of microfluidics, and particularly relates to a self-driven disc type blood separation and detection device and method. BACKGROUND

[0002] Blood is composed of blood cells and plasma, and is a treasure house of human body function information, and is closely related to the tissues and organs of each system of the human body. The types and contents of biomarkers in blood can reflect the health status of the body. Therefore, blood detection is also one of the most common detection methods in our daily life.

[0003] Generally, blood detection needs to filter out blood cells in blood through centrifugation, sedimentation and the like, so as to avoid the interference of blood cells on subsequent detection. However, these traditional blood separation methods need to be carried out in a laboratory, and depend on large centrifugal equipment, and need more blood samples, which is easy to cause waste. Moreover, the traditional centrifugal equipment is too large in size, and is not convenient to carry and carry, and cannot meet the needs of timely detection. The time interval between plasma separation and analysis is usually relatively long, and the separation time of most centrifuges is about 1 h, which will affect the accuracy and consistency of plasma component analysis to some extent, and it is difficult to meet the needs of real-time and accurate detection of blood components. Moreover, most blood detection fees are too high, which restricts the development of disease blood census detection. SUMMARY

[0004] The application aims to solve the problems in the background art, and provides a self-driven disc type blood separation and detection device and method.

[0005] In order to achieve the above-mentioned purpose, the application adopts the following technical scheme: a self-driven disc type blood separation and detection device, comprising: a sample adding layer, a separation layer and a detection layer, the sample adding layer, the separation layer and the detection layer are connected with each other in sequence from top to bottom by screws.

[0006] The sample adding layer is provided with a sample adding port.

[0007] The separation layer is provided with four repeated separation units, each separation unit comprises a sample inlet, a microfluidic channel, a filter groove, a sample outlet and a waste liquid collection chamber, the sample inlet, the filter groove, the sample outlet and the waste liquid collection chamber are connected in sequence through the microfluidic channel, the sample inlet and the waste liquid collection chamber are arranged at two ends of the microfluidic channel, the inlet of the sample inlet and the outlet of the sample adding port are overlapped and connected, and the filter groove and the sample outlet are arranged at the bottom end of the microfluidic channel.

[0008] The detection layer is provided with a screen printing electrode interface, the screen printing electrode interface is provided with a screen printing electrode, and the screen printing electrode is located at the outlet of the sample outlet.

[0009] Further, the screen-printed electrode interface is connected with the outlet of the sample outlet, the size of the screen-printed electrode interface matches the size of the outlet of the sample outlet, and the screen-printed electrode interface is attached to the outlet of the sample outlet.

[0010] Further, the sample adding layer above the filter groove is coated with a hydrophobic pattern layer, and the size of the hydrophobic pattern layer matches the size of the groove of the filter groove.

[0011] Further, the separation layer is in a low-pressure state before detection.

[0012] Further, the four corners of the sample adding layer are provided with first screw holes, the four corners of the separation layer are provided with second screw holes, and the four corners of the detection layer are provided with third screw holes, the diameters of the first screw holes, the second screw holes and the third screw holes are 1mm, and the positions of the first screw holes, the second screw holes and the third screw holes overlap with each other.

[0013] Further, the sample adding layer and the detection layer are both circular glass slides with a diameter of 50mm and a thickness of 2mm.

[0014] Further, the sample adding port has a diameter of 5mm and a depth of 2mm, and the sample adding port is located at the center of the sample adding layer.

[0015] Further, the separation layer is a circular PDMS with a diameter of 50mm and a thickness of 3mm.

[0016] Further, the sample inlet is a small hole with a diameter of 5mm and a depth of 300μm, the microfluidic channel has a width of 2mm and a depth of 300μm, the filter groove is a cylindrical micro-well with a diameter of 2mm and a height of 2mm, the sample outlet is a small hole with a diameter of 4mm and a depth of 3mm, and the waste liquid collection chamber is a cuboid with a length of 2mm, a width of 2mm and a depth of 300μm.

[0017] The application also provides a detection method of the self-driven disc type blood separation and detection device, comprising:

[0018] Step 1: first determine the measured object, and modify the screen-printed electrode based on the characteristics of the measured object;

[0019] Step 2: first store the separation layer PDMS chip in a vacuum for more than 15 minutes, so that the microfluidic channel is in a negative pressure state;

[0020] Step 3: take out the PDMS chip, and fix the sample adding layer, the separation layer and the detection layer in sequence by using screws;

[0021] Step 4: inject 5ml of whole blood sample from the sample inlet;

[0022] Step 5: under the negative pressure driving of the device, the blood sample starts to flow along the microfluidic channel;

[0023] Step 6: when the blood sample flows through the filter groove, due to the difference in gravity suffered by the blood cells and the plasma, the heavier blood cells drop into the filter groove, and the lighter plasma continues to flow forward;

[0024] Step 7: the separated blood sample reaches the sample outlet, and the screen-printed electrode is connected below the sample outlet; when the plasma completely covers the three-electrode area of the screen-printed electrode, the electrochemical workstation is started to detect;

[0025] Step 8: according to different detection types, different detection modes of the electrochemical workstation are selected, and the results given by the electrochemical workstation are analyzed, so that the blood detection purpose is achieved.

[0026] Beneficial effects:

[0027] 1. The blood sample can be separated and detected quickly and efficiently, and external power supply is not needed, so that the operation steps are optimized, the sample pollution is reduced, and the blood detection cost is greatly reduced.

[0028] 2. The blood sample can be detected in time.

[0029] 3. The application can detect multiple disease markers at the same time, and the detection efficiency is improved.

[0030] 4. The application has expandability, and more samples can be detected by increasing the blood separation unit in the disc structure.

[0031] 5. The application uses a screen-printed electrode instead of a traditional three-electrode, which can realize plug and play, greatly simplifies the detection process, and saves cost. DETAILED DESCRIPTION

[0032] Figure 1 It is a sampling layer structure schematic diagram of the embodiment of the application;

[0033] Figure 2 It is a separation layer top view of the embodiment of the application;

[0034] Figure 3 It is a separation unit section view of the embodiment of the application;

[0035] Figure 4 It is a detection layer structure schematic diagram of the embodiment of the application;

[0036] Figure 5 It is a screen-printed electrode structure schematic diagram of the embodiment of the application.

[0037] In the figure: 1, sample layer, 1.1, sample port, 1.2, first screw hole, 1.3, hydrophobic layer, 2, separation layer, 2.1, sample inlet, 2.2, microfluidic channel, 2.3, filter groove, 2.4, sample outlet, 2.5, waste liquid collection chamber, 2.6, second screw hole, 3, detection layer, 3.1, screen printed electrode interface, 3.2, third screw hole, 4, screen printed electrode, 4.1, reference electrode, 4.2, counter electrode, 4.3, working electrode, 4.4, working electrode lead, 4.5, reference electrode lead, 4.6, counter electrode lead. DETAILED DESCRIPTION

[0038] The application will be further explained below with reference to the accompanying drawings.

[0039] Reference Figures 1-4 The embodiment discloses a self-driven disc type blood separation and detection device, which comprises a sample layer 1, a separation layer 2 and a detection layer 3, and the sample layer 1, the separation layer 2 and the detection layer 3 are sequentially connected with each other from top to bottom, wherein the connection mode between the sample layer 1, the separation layer 2 and the detection layer 3 can adopt a mode of mutual connection by means of screws and bolts.

[0040] The sample layer 1 is provided with a sample port 1.1 and a first screw hole 1.2.

[0041] The separation layer 2 is provided with four repeated separation units and a second screw hole 2.6, and each separation unit comprises a sample inlet 2.1, a microfluidic channel 2.2, a filter groove 2.3, a sample outlet 2.4 and a waste liquid collection chamber 2.5. The sample inlet 2.1, the filter groove 2.3, the sample outlet 2.4 and the waste liquid collection chamber 2.5 are sequentially connected in communication through the microfluidic channel 2.2. The sample inlet 2.1, the microfluidic channel 2.2, the filter groove 2.3, the sample outlet 2.4 and the waste liquid collection chamber 2.5 are photoetched on the upper surface of the separation layer 2, and the upper portions of the sample inlet 2.1, the microfluidic channel 2.2, the filter groove 2.3, the sample outlet 2.4 and the waste liquid collection chamber 2.5 are all open. The sample layer 1 is attached and covers the upper surface of the separation layer 2, and seals the upper openings of the microfluidic channel 2.2, the filter groove 2.3, the sample outlet 2.4 and the waste liquid collection chamber 2.5. The sample inlet 2.1 and the waste liquid collection chamber 2.5 are arranged at two ends of the microfluidic channel 2.2, the inlet of the sample inlet 2.1 and the outlet of the sample port 1.1 are mutually overlapped and connected in communication, and the filter groove 2.3 and the sample outlet 2.4 are arranged at the bottom end of the microfluidic channel 2.2.

[0042] The detection layer 3 is provided with a screen printed electrode interface 3.1 and a third screw hole 3.2, the screen printed electrode interface 3.1 is installed with a screen printed electrode 4, and the screen printed electrode 4 is located at the outlet of the sample outlet 2.4.

[0043] In this embodiment, the first screw hole 1.2, the second screw hole 2.6 and the third screw hole 3.2 are all 1mm in diameter and are coincident with each other, and can be fixed by screws during assembly to make the sample layer 1, the separation layer 2 and the detection layer 3 tightly adhere to each other, wherein the sample layer 1 and the detection layer 3 are made of glass slides, and the separation layer 2 is made of PDMS.

[0044] Reference Figures 1-3 The sample layer 1 is provided with a hole with a diameter of 5mm and a depth of 2mm at the center, and the sample layer 1 is placed on the top of the sample inlet 2.1, the microfluidic channel 2.2, the filter groove 2.3, the sample outlet 2.4 and the waste liquid collection chamber 2.5, and the sample layer 1 is tightly combined with the separation layer 2 in the device by screws, so as to prevent the blood sample from being contaminated by external factors, as the sampling port 1.1 of the blood sample, and coincides with the sample inlet 2.1 in the separation layer 2, and the sample layer 1 above the filter groove 2.3 is coated with a hydrophobic layer 1.3, and the size of the hydrophobic layer 1.3 matches the size of the filter groove 2.3, so as to prevent the blood cells in the filter groove 2.3 from overflowing and affecting the detection result.

[0045] In this embodiment, the sample layer 1 is provided with a sample port for collecting blood samples, and covering the separation layer can also prevent the pollution of the external environment, and the sample layer 1 is coated with a hydrophobic material at the overlapping position above the filter groove 2.3, so that the filter groove 2.3 can fully filter the blood cells, and thus the detection result is more accurate.

[0046] Reference Figures 2-3, the separation layer 2 is provided with four repeated separation units and screw holes 2.6, each separation unit has a sample inlet 2.1, a microfluidic channel 2.2, a filter groove 2.3, a sample outlet 2.4 and a waste liquid collection chamber 2.5; before starting work, the separation layer 2 needs to be placed in a low pressure condition (<0.3 atm) for more than 15 min, then taken out for assembly and fixed with screws; the user drops the blood sample into the sampling port 1.1 with a diameter of 5 mm and a depth of 2 mm, then sends it to the sample inlet 2.1 with a diameter of 5 mm and a depth of 300 μm, and waits for further operation; the PDMS stores a certain potential in a low pressure state, when it comes into contact with the outside world, due to the difference from atmospheric pressure, energy is generated to push the blood along the microfluidic channel 2.2 with a width of 2 mm and a depth of 300 μm, then the blood sample is separated by using the different masses of blood cells and plasma, the blood cells with larger weight are left in the cylindrical filter groove 2.3 with a diameter of 2 mm and a depth of 2 mm, and the plasma continues to advance to the sample outlet 2.4 with a diameter of 4 mm and a depth of 3 mm (the diameter of the sample outlet changes according to the screen printed electrode three-electrode area, and can cover the three-electrode area); the screen printed electrode 4 is connected below the sample outlet 2.4, when the plasma falls on the screen printed electrode 4, electrochemical detection can be carried out; the waste liquid collection chamber 2.5 with a length of 2 mm, a width of 2 mm and a depth of 300 μm is connected behind the sample outlet 2.4, which collects the excess blood sample and stops flowing before the filter groove 2.3 is full.

[0047] In this embodiment, the separation layer 2 needs to be in a low pressure state before detection (time > 15 min, pressure < 0.3 atm), so that the PDMS material stores energy, and the separation layer 2 has four repeated separation units, which can realize multi-channel detection at the same time and improve the detection efficiency. The sample inlet 2.1 is used to store the blood sample, the filter groove 2.3 is used to filter the blood cells in the blood sample, the sample outlet 2.4 is used to send the filtered blood directly to the screen printed electrode for detection, the waste liquid collection chamber 2.5 is used to collect the excess blood sample and stop flowing before the filter groove 2.3 is full, preventing the blood cells from overflowing and affecting the detection.

[0048] Reference Figure 4The detection layer 3 is provided with a screen-printed electrode interface 3.1, and a screen-printed electrode 4 is installed on the screen-printed electrode interface 3.1. The screen-printed electrode 4 is located at the outlet of the sample outlet 2.4. The screen-printed electrode interface 3.1 is in communication with the outlet of the sample outlet 2.4, and the size of the screen-printed electrode interface 3.1 matches the size of the outlet of the sample outlet 2.4. The screen-printed electrode interface 3.1 is attached to the outlet of the sample outlet 2.4, so that the screen-printed electrode interface 3.1 is located corresponding to the sample outlet 2.4. The length and depth of the screen-printed electrode interface 3.1 are determined according to the selected screen-printed electrode 4. The above arrangement ensures that the screen-printed electrode 4 can be inserted into the detection layer, and the three-electrode area of the screen-printed electrode is opposite to the sample outlet 2.4. The inserted screen-printed electrode 4 can seal the blood in the sample outlet 2.4, preventing the blood sample from flowing out. The detection layer 3 also plays a supporting and sealing role in the device.

[0049] In the embodiment, the detection layer 3 is located at the lowermost layer of the self-driven disc type blood separation and detection device, and plays a supporting role while preventing the blood sample from flowing out.

[0050] Reference Figure 5 The screen-printed electrode 4 includes a reference electrode 4.1, a counter electrode 4.2, a working electrode 4.3, a working electrode lead 4.4, a reference electrode lead 4.5, and a counter electrode lead 4.6. The plasma in the sample outlet 2.4 needs to completely cover the reference electrode 4.1, the counter electrode 4.2, and the working electrode 4.3, so that electrochemical detection can be performed. The working electrode lead 4.4, the reference electrode lead 4.5, and the counter electrode lead 4.6 are respectively connected to corresponding positions of an electrochemical workstation. After the electrochemical workstation is set, detection is performed.

[0051] The embodiment of the application can quickly and efficiently separate a small amount of blood sample and realize multi-channel electrochemical detection, thereby improving the detection efficiency of the blood sample, reducing the volume, simplifying the structure, being easy to carry, saving time, and reducing cost. The embodiment of the application can completely meet the requirements of rapid and efficient blood separation and detection. The self-driven disc type blood separation and detection device separates and detects the blood sample without manual intervention. The device automatically operates as soon as the blood sample is dropped, and does not need external power supply. The complexity of operation is greatly reduced, and the pollution of the sample is also reduced. The device can output results in about ten minutes, thereby greatly shortening the detection time. The device does not need to use professional equipment, and the use cost is reduced.

[0052] The application does not need external power supply, thereby reducing the sample operation and improving the efficiency of blood separation. The blood separation and detection operations are integrated, thereby reducing the volume of the device and realizing instant and multi-channel detection of the whole blood sample. In addition, the screen-printed electrode is connected to the interface, thereby replacing the traditional three-electrode and realizing plug-and-play.

[0053] The embodiment also provides a detection method of the self-driven disc type blood separation and detection device, comprising the following steps.

[0054] Step 1: first determine the to-be-tested substance, and modify the screen-printed electrode 4 according to the to-be-tested substance.

[0055] Step 2: first store the separation layer 2 in a low-pressure state (<0.3 atm) for more than 15 min, so that the inside of the separation layer is in a low-pressure state.

[0056] Step 3: take out the separation layer 2 from the low-pressure environment, and fix the sampling layer 1, the separation layer 2 and the detection layer 3 in sequence by using a screw.

[0057] Step 4: inject 5 ml of a whole blood sample from the sampling port 1.1 into the sample inlet 2.1.

[0058] Step 5: due to the pressure difference between the inside and outside of the separation layer, the blood sample is driven to flow along the microfluidic channel 2.2.

[0059] Step 6: when the blood sample flows through the filter groove 2.3, due to the difference in gravity suffered by the blood cells and the plasma, the heavier blood cells drop into the filter groove 2.3, and the lighter plasma continues to flow forward.

[0060] Step 7: the separated blood sample reaches the sample outlet 2.4, and the screen-printed electrode 4 is connected below the sample outlet 2.4; when the blood completely covers the three-electrode area of the screen-printed electrode 4, the electrochemical workstation is started to detect.

[0061] Step 8: different detection methods of the electrochemical workstation are selected according to different detection types, and the blood detection purpose is achieved by analyzing the results given by the electrochemical workstation.

[0062] The above only describes the preferred embodiments of the present application, and it should be noted that, for those skilled in the art, some improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A self-driven disc type blood separation and detection device, characterized in that, The utility model relates to a kind of microfluidic chip, including: Sample layer (1), separation layer (2) and detection layer (3), the sample layer (1), separation layer (2) and detection layer (3) are sequentially connected from top to bottom with each other; The sample layer (1) is provided with sample port (1.1);The four corners of the sample layer (1) are provided with first screw hole (1.2), the four corners of the separation layer (2) are provided with second screw hole (2.6), the four corners of the detection layer (3) are provided with third screw hole (3.2), the diameter of the first screw hole (1.2), second screw hole (2.6) and third screw hole (3.2) is 1mm, and the first screw hole (1.2), second screw hole (2.6) and third screw hole (3.2) are overlapped with each other; The separation layer (2) is provided with four repeated separation units, each separation unit includes sample inlet (2.1), microfluidic channel (2.2), filter groove (2.3), sample outlet (2.4) and waste liquid collection chamber (2.5), and is opened on the upper surface of separation layer (2), the sample inlet (2.1), filter groove (2.3), sample outlet (2.4) and waste liquid collection chamber (2.5) are sequentially communicated by microfluidic channel (2.2), the sample inlet (2.1) and waste liquid collection chamber (2.5) are arranged at both ends of microfluidic channel (2.2), the inlet of the sample inlet (2.1) and the outlet of the sample port (1.1) are overlapped and connected, the filter groove (2.3) and the sample outlet (2.4) are arranged at the bottom end of the microfluidic channel (2.2); The separation layer (2) is in a low pressure state of <0.3atm before detection; The detection layer (3) is provided with screen printing electrode interface (3.1), the screen printing electrode interface (3.1) is installed with screen printing electrode (4), the screen printing electrode (4) is located at the outlet of sample outlet (2.4);The screen printing electrode interface (3.1) is communicated with the outlet of sample outlet (2.4), the size of the screen printing electrode interface (3.1) matches the size of the outlet of sample outlet (2.4), and the screen printing electrode interface (3.1) is attached to the outlet of sample outlet (2.4);The sample layer (1) directly above the filter groove (2.3) is coated with hydrophobic layer (1.3), and the size of the hydrophobic layer (1.3) matches the size of the slot of the filter groove (2.3).

2. The self-driven disc type blood separation and detection device according to claim 1, wherein, The sample layer (1) and detection layer (3) are both circular glass slides with a diameter of 50mm and a thickness of 2mm.

3. The self-driven disc type blood separation and detection device according to claim 1, wherein, The diameter of the sample port (1.1) is 5mm, and the depth is 2mm, and the sample port (1.1) is located at the center of the sample layer (1).

4. The self-driven disc type blood separation and detection device according to claim 1, wherein, The separation layer (2) is a circular PDMS with a diameter of 50mm and a thickness of 3mm.

5. The self-driven disc type blood separation and detection device according to claim 1, wherein, The sample inlet (2.1) is a small hole with a diameter of 5mm and a depth of 300μm, the microfluidic channel (2.2) is 2mm wide and 300μm deep, the filter groove (2.3) is a cylindrical micro-well with a diameter of 2mm and a height of 2mm, the sample outlet (2.4) is a small hole with a diameter of 4mm and a depth of 3mm, and the waste liquid collection chamber (2.5) is a cuboid with a length of 2mm, a width of 2mm, and a depth of 300μm.

6. A detection method based on the self-driven disc type blood separation and detection device of claim 1, characterized in that, Comprise: Step 1: First determine the test substance, and modify the screen-printed electrode based on the characteristics of the test substance; Step 2: First, store the separation layer PDMS chip in a vacuum for more than 15 minutes to make the microfluidic channel inside in a negative pressure state; Step 3: Take out the PDMS chip and use screws to fix the sample layer, separation layer and detection layer in order; Step 4: Inject 5ml of whole blood sample from the sample inlet; Step 5: Under the driving of the negative pressure inside the device, the blood sample begins to flow along the microfluidic channel; Step 6: When the blood sample flows through the filter groove, due to the difference in gravity between blood cells and plasma, the heavier blood cells fall into the filter groove, while the lighter plasma continues to flow forward; Step 7: After separation, the blood sample reaches the sample outlet, which is connected to the screen-printed electrode. When the plasma completely covers the three-electrode area of the screen-printed electrode, turn on the electrochemical workstation for detection; Step 8: According to different detection types, select different detection methods of the electrochemical workstation, and analyze the results given by the electrochemical workstation to achieve the purpose of blood detection.

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