CN108212228B - Whole blood separation micro-fluidic chip, detection device and whole blood detection method - Google Patents
Whole blood separation micro-fluidic chip, detection device and whole blood detection method Download PDFInfo
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Abstract
The invention discloses a whole blood separation micro-fluidic chip, a detection device and a whole blood detection method, and belongs to the technical field of medical appliances. The detection device is provided with the microfluidic chip. The microfluidic chip has a simple structure, is convenient to manufacture and is suitable for market popularization. The microfluidic chip can integrate the whole blood separation process, the serum and diluent mixing process and the antibody antigen specific binding process into the same chip, and can detect a plurality of indexes on the same chip. According to the detection device and the detection method provided by the invention, the LSPR effect of the nano particles is utilized in the test process, so that the concentration of the object to be detected can be judged by detecting the absorbance change condition without marking and without pollution in real time and with high sensitivity.
Description
Technical Field
The invention relates to the technical field of medical instruments, in particular to a whole blood separation micro-fluidic chip, a detection device and a detection method.
Background
Microfluidic is a technology for precisely controlling and manipulating microscale fluids, in particular to submicron structures. In the 80 s of the 20 th century, microfluidic technology began to rise, and has been developed in the directions of DNA chips, lab-on-a-chip, micro-sampling technology, micro-thermal technology, and the like.
Microfluidic chips, originally called "lab-on-a-chips" in the united states and "micro-integrated analytical chips" (micrototal analytical systems) in europe, are the main platform for microfluidic technology (Microfluidics) implementation, and can integrate basic operation units of sample preparation, reaction, separation, detection, etc. in biological, chemical and medical analysis processes into a chip with a micrometer scale, and automatically complete the whole analysis process. Microfluidic chips which have the advantages of light volume, small sample and reagent amount, high reaction speed, capability of being processed in a large number of parallel ways, disposability and the like, have great potential in the fields of biology, chemistry, medicine and the like, and have been developed into a brand-new research field of biology, chemistry, medicine, fluid, electronics, materials, machinery and the like.
At present, microfluidic chips are commonly used for separating serum or plasma from whole blood, and many schemes adopt a centrifugal technology or a filter layer for separation, quantification and distribution. Serum is prepared and then tested basically based on ELISA (enzyme-linked immunosorbent assay, enzyme-linked immunosorbent assay, ELISA) or chemiluminescence, and the traditional techniques have complicated processes, require horseradish enzyme markers or chemiluminescent reagents, have high cost and long time consumption, and are not suitable for rapid detection in families or communities.
The Chinese patent with the patent number ZL201310442679.5 discloses a centrifugal whole blood analysis micro-fluidic chip, but the structure is complex, the manufacturing process is too complicated, and the chip is not suitable for mass popularization and application, so that the quick detection requirement of families or communities cannot be met.
Disclosure of Invention
The invention aims to provide a whole blood separation microfluidic chip which has a simple structure, is convenient to manufacture and is suitable for market popularization, wherein a serum dilution mixing process and a protein adsorption detection process to be detected are concentrated on one chip, and a plurality of objects to be detected can be detected simultaneously by the same chip.
To achieve the purpose, the invention adopts the following technical scheme:
a whole blood separation microfluidic chip comprising:
a base;
a main channel disposed on the substrate, at least one set of functional units, and a sample well and a blood cell collection well in communication through the main channel;
the functional unit includes:
the serum collecting hole, the mixing hole and the testing hole are communicated in sequence, and the serum collecting hole is communicated with the main channel;
a diluent hole in communication with the mixing hole;
and the waste liquid hole is communicated with the test hole.
The whole blood separation microfluidic chip comprises a centrifuge and a matrix connected with the centrifuge, wherein a main channel, at least one group of functional units, a sample hole and a blood cell collecting hole which are communicated with each other through the main channel are arranged on the matrix;
the functional unit includes:
the serum collecting hole, the mixing hole and the testing hole are communicated in sequence, and the serum collecting hole is communicated with the main channel;
a diluent hole in communication with the mixing hole;
a waste well in communication with the test well;
the test hole penetrates through the substrate, and a transparent substrate for placing the detection probe and the nano particles is arranged at the bottom of the test hole.
Preferably, the substrate is a circular or rectangular light transmissive sheet.
Preferably, the matrix is made of polymethyl methacrylate (PMMA for short).
Preferably, each of the primary channel, sample well, blood cell collection well and functional unit extends through the matrix.
Preferably, each hole, the sample hole and the blood cell collecting hole in the functional unit are all round holes, and the pore diameters of the holes can be equal or unequal.
Preferably, the transparent substrate is any one of a glass sheet, a quartz sheet, a transparent ceramic sheet and a transparent polymer sheet.
Preferably, the nanoparticle is any one of pure gold, pure silver and pure platinum, or an alloy containing at least one of gold, silver and platinum.
The whole blood separation micro-fluidic chip provided by the scheme has a simple structure, is convenient to manufacture and is suitable for market popularization. The microfluidic chip integrates the serum separation process, the serum and diluent mixing process and the antibody antigen specific binding process of whole blood into the same chip, and can detect a plurality of indexes on the same chip. The testing process utilizes the LSPR effect (Localized Surface Plasmon Resonance, local surface plasmon resonance effect, LSPR effect for short) of the nano particles, can realize the detection without marking and with no pollution, real time and high sensitivity, and judges the concentration of the to-be-detected object by detecting the absorbance change condition.
In a preferred scheme, a positioning hole is added on the substrate above the structure of the whole blood separation micro-fluidic chip, and the positioning hole can be positioned at the center of the substrate, preferably, a round hole, a square hole or a diamond hole. Further preferably, the locating hole extends through the base body. The positioning holes are arranged to facilitate quick positioning of the microfluidic chip when the microfluidic chip is mounted on the centrifugal machine, and prevent the microfluidic chip from shifting in the centrifugal process, so that the microfluidic chip is ensured to shake stably in the whole centrifugal process.
In a further preferred embodiment, the number of functional units is one to ten, that is to say the number of functional units can be one, two, three, four, five, six, seven, eight, nine or ten, in order to meet different detection requirements.
In order to save space and improve the compactness of the whole blood separation microfluidic chip, a preferred scheme is that the main channel is annular, such as circular, elliptical or rectangular, so that the functional unit can be arranged around the main channel, and the structure is more compact. Preferably, the functional units are four groups and are distributed in a cross shape around the annular main channel.
In still another preferred embodiment, the number of the dilution holes in each of the functional units is at least one, and the specific number is set according to the concentration of the analyte.
The whole blood separation microfluidic chip is preferably arranged on the detection device for testing, so that the test hole is close to the edge of the substrate. In order to facilitate the inflow and outflow of the mixed solution and the reaction with the antibody serving as the detection probe, preferably, the hole wall of the test hole is provided with a sample inlet and a liquid outlet which are opposite to each other, the sample inlet is close to the top of the test hole, and the liquid outlet is close to the bottom of the test hole.
The invention also aims to provide the whole blood separation micro-fluidic chip which has a simple structure, is convenient to manufacture and is suitable for market popularization, and can rapidly realize the test of blood on the micro-fluidic chip.
To achieve the purpose, the invention adopts the following technical scheme:
the whole blood separation microfluidic chip comprises a centrifuge and a matrix connected with the centrifuge, wherein a test hole penetrating through the matrix is formed in the matrix, and a transparent substrate for placing a detection probe and nano particles is arranged at the bottom of the test hole.
Preferably, the transparent substrate is any one of a glass sheet, a quartz sheet, a transparent ceramic sheet and a transparent polymer sheet.
Preferably, the nanoparticle is any one of pure gold, pure silver and pure platinum, or an alloy containing at least one of gold, silver and platinum.
In a preferred scheme, a positioning hole is added on the substrate above the structure of the whole blood separation micro-fluidic chip, and the positioning hole can be positioned at the center of the substrate, preferably, a round hole, a square hole or a diamond hole. Further preferably, the locating hole extends through the base body. The positioning holes are arranged to facilitate quick positioning of the microfluidic chip when the microfluidic chip is mounted on the centrifugal machine, and prevent the microfluidic chip from shifting in the centrifugal process, so that the microfluidic chip is ensured to shake stably in the whole centrifugal process.
The whole blood separation microfluidic chip is preferably arranged on the detection device for testing, so that the test hole is close to the edge of the substrate. In order to facilitate the inflow and outflow of the mixed solution and the reaction with the antibody serving as the detection probe, preferably, the hole wall of the test hole is provided with a sample inlet and a liquid outlet which are opposite to each other, the sample inlet is close to the top of the test hole, and the liquid outlet is close to the bottom of the test hole.
As a preferred embodiment, the centrifuge is a multistage variable speed centrifuge.
Specifically, the centrifuge can adjust different rotating speeds at different stages of whole blood immunodetection so as to meet the processing requirements of detection samples at different stages.
The invention also aims to provide a detection device and a whole blood detection method provided with the whole blood separation microfluidic chip, and the detection process utilizes the characteristic of no marking of the LSPR technology, so that the whole detection process is simple and convenient to operate, short in time and low in cost, and the rapid detection requirement of families or communities is met.
To achieve the purpose, the invention adopts the following technical scheme:
a detection device comprising a microfluidic chip, the microfluidic chip being a whole blood separation microfluidic chip as described in any one of the above; one end of a test hole in the whole blood separation microfluidic chip is provided with a parallel light component, and the other end is provided with a spectrum analysis component.
As a preferable technical scheme, the parallel light assembly comprises a light source, an incoming optical fiber and an incoming collimator, wherein the light source is connected with the incoming collimator through the incoming optical fiber, the incoming collimator is arranged at one end of the test hole far away from the spectrum analysis assembly, and the emission direction of the incoming collimator is aligned with the test hole.
In particular, the light source is for emitting continuous visible white light in a wavelength range of 380nm to 800nm, and the light source may be a halogen lamp.
Specifically, the incoming optical fiber is used as a carrier for light transmission, so that light is transmitted from the light source to the incoming collimator.
Specifically, the incoming collimator is used for preparing parallel light rays and emitting the parallel light rays to the test hole.
As a preferred technical solution, the spectrum analysis component includes a spectrometer, an outgoing optical fiber and an outgoing collimator, the spectrometer is connected with the outgoing collimator through the outgoing optical fiber, the outgoing collimator is installed at one end of the test hole far away from the parallel light component, and the receiving direction of the outgoing collimator is aligned with the test hole.
Specifically, when the parallel light assembly emits light, the incoming collimator is aligned with the test hole, the outgoing collimator is aligned with the test hole, and the axis of the incoming collimator, the axis of the outgoing collimator and the axis of the test hole are positioned on the same straight line.
Preferably, the incoming collimator is located directly above the test aperture and the outgoing collimator is located directly below the test aperture.
Preferably, the incoming collimator is located directly below the test aperture and the outgoing collimator is located directly above the test aperture.
Specifically, the outgoing collimator is configured to receive light after passing through the test hole.
Specifically, the outgoing optical fiber is used as a carrier for light transmission, so that light is transmitted from the outgoing collimator to the spectrometer.
Specifically, the spectrometer analyzes the output signal of the outgoing optical fiber to form a spectrum analysis result, and provides data support for subsequent body health analysis.
The invention also provides a whole blood detection method, which comprises the following steps:
dripping blood into a sample hole of a whole blood separation microfluidic chip, connecting the microfluidic chip with a centrifuge, starting the centrifuge, enabling blood cells to enter a blood cell collecting hole, enabling serum to enter a serum collecting hole and flowing into a mixing hole;
the diluent in the diluent hole enters the mixing hole to be mixed with the serum, the mixed liquid flows into the testing hole, and the protein to be tested in the serum is specifically combined with the detection probe;
the mixed liquid in the test hole flows into the waste liquid hole;
absorbance testing was performed on the test wells.
Compared with the traditional detection method, the detection device and the detection method have the advantages that whole blood such as fingertip blood or venous blood can be directly added into the microfluidic chip, and serum is obtained by separating the whole blood through the centrifuge with the small size and then distributed to the test holes. The whole process is simple to operate, has low professional requirements on users, does not need to prepare serum or plasma in advance, and only needs to know how to take fingertip blood or venous blood for the users.
The principle of the scheme is based on the local surface plasmon resonance effect (Localized Surface Plasmon Resonance, LSPR effect for short) of the nano gold particles, and the principle is most remarkable in that the previous methods such as an enzyme-linked immunosorbent assay or radiolabeling and the like are avoided, and the operations such as an antibody for catalyzing enzyme, a chromogenic agent or a radioisotope and the like are required to be added in the test process. The whole use process of utilizing this scheme to detect need not to add any marker, after adding whole blood detection sample, detect the disease marker in the sample and can combine with the antibody that is the detection probe in the test hole of micro-fluidic chip, arouse the environmental change around the nano gold particle to lead to the nano gold particle to change to the absorption of light, consequently through the change of analysis absorption peak, can directly test the concentration of antibody, it is based on such principle for the test process is simple and convenient, the consumptive material that the test needs is less, thereby realize the cost reduction.
Specifically, the chip based on the LSPR effect can directly realize qualitative and quantitative measurement of the analyte by utilizing the specific reaction of the analyte in the detection process. When a certain analyte is to be detected, a substance capable of specifically binding with the corresponding analyte can be firstly used as a detection probe to be modified on the surface of the nano gold particles or between the nano gold particles, a sample containing the analyte can be specifically bound with the detection probe after entering the test hole, and the detection control system can analyze whether the sample contains the analyte or the amount of the analyte according to the extinction change of a chip provided with the nano gold.
Currently, chips are used for detection of protein-protein or protein-DNA interactions, chain coccus-biotin reactions, immunoglobulin G (IgG) tests or antigen-antibody interactions, where protein antigen antibodies can be used to detect various diseases, and in disease applications, disease markers can be detected to determine whether a disease is present.
The disease markers in the disease detection comprise tumors, myocardial infarction, liver inflammation, infection, cerebral infarction, immune function and the like, wherein the tumors specifically comprise lung cancer, liver cancer, gastric cancer, pancreatic cancer, intestinal cancer, breast cancer, prostate cancer and the like; the myocardial infarction comprises troponin I, troponin T, myoglobin, creatine hormone isozymes, N-terminal B-type natriuretic peptide precursors and the like; liver inflammation includes hepatitis b virus antibody and hepatitis c virus core antigen; the infection includes C-reactive protein, procalcitonin, interleukin 6, etc.
Different types of diseases have different specific disease markers, and when it is desired to detect a certain disease, an antibody that specifically binds to a specific disease marker can be used as a detection probe. For example, when detecting liver cancer, it is necessary to detect alpha-fetoprotein (alpha FP or AFP) as a disease marker, and AFP antibody can be used as a detection probe; when the tumor necrosis factor TNF-a of a disease marker needs to be detected, adalimumab can be used as a detection probe; when detecting hepatitis B, it is necessary to detect hepatitis B surface antigen (HBsAg) which is a disease marker, and the hepatitis B surface antibody (HBsAb) can be used as a detection probe; when the disease marker hepatitis B e antigen (HBeAg) needs to be detected, a hepatitis B e antibody (HBeAb) can be used as a detection probe; when detecting breast cancer, the disease marker human breast cancer CA15-3 needs to be detected, and a human breast cancer CA153 antibody can be used as a detection probe; in the detection of herpes virus infection, it is necessary to detect the disease marker Epstein-Barr virus (EBV), and an EBV-CA (IgM) antibody can be used as a detection probe. Therefore, a plurality of test holes are formed in the chip, and different antibodies are modified on the nanogold of different test holes to serve as detection probes, so that a plurality of different diseases can be detected simultaneously, the detection is more efficient and convenient, and the kit is suitable for families, communities and hospitals.
The invention has the beneficial effects that:
the whole blood separation micro-fluidic chip provided by the invention has a simple structure, is convenient to manufacture and is suitable for market popularization. The microfluidic chip integrates the serum separation process, the serum and diluent mixing process and the antibody antigen specific binding process of whole blood into the same chip, and can detect a plurality of indexes on the same chip. The detection device and the whole blood detection method provided by the invention can realize pollution-free, real-time and high-sensitivity detection without marking by utilizing the LSPR effect (Localized Surface Plasmon Resonance, local surface plasmon resonance effect, LSPR effect for short) of the nano particles in the test process, and judge the concentration of the to-be-detected object by detecting the absorbance change condition.
Drawings
Fig. 1 is a top view of a whole blood separation microfluidic chip provided by the present invention;
FIG. 2 is a partial view of a test well in a whole blood separation microfluidic chip provided by the present invention;
fig. 3 is a schematic diagram of a detection device provided by the present invention.
In the figure: a-substrate, 1-main channel, 2-serum collection well, 3-dilution well, 4-waste well, 5-sample well, 6-positioning well, 7-blood cell collection well, 8-mixing well, 9-test well, 10-sample inlet, 11-hole wall, 12-transparent substrate, 13-detection probe, 14-nanoparticle, 15-liquid outlet, 20-centrifuge, 31-light source, 32-incoming optical fiber, 33-incoming collimator, 41-spectrometer, 42-outgoing optical fiber, 43-outgoing collimator, 50-human-computer interaction device.
Detailed Description
The technical scheme of the invention is further described below by the specific embodiments with reference to the accompanying drawings.
Example 1
As shown in fig. 1, a whole blood separation microfluidic chip includes a substrate a, a main channel 1 provided on the top surface of the substrate a, at least one set of functional units, and a sample well 5 and a blood cell collection well 7 communicating through the main channel 1. Wherein, the matrix a is a round light-transmitting sheet and is made of polymethyl methacrylate (PMMA for short). The main channel 1 is in a circular ring shape and is coaxial with the matrix a so as to save space, improve the compactness of the whole blood separation microfluidic chip and facilitate the arrangement of a plurality of groups of functional units. In this embodiment, the functional units are four groups and are arranged in a cross shape around the circular main channel 1. The sample well 5 and the blood cell collection well 7 are in communication via a first auxiliary channel, and the first auxiliary channel intersects the main channel 1, thereby enabling the sample well 5 and the blood cell collection well 7 to be in communication via the main channel 1.
As shown in fig. 1, the functional units include a serum collection well 2, a diluent well 3, a waste well 4, a mixing well 8, and a test well 9. The serum collecting hole 2 is directly communicated with the main channel 1, the serum collecting hole 2 is communicated with the mixing hole 8 through the second auxiliary channel, the mixing hole 8 is communicated with the test hole 9 through the third auxiliary channel, furthermore, the diluent hole 3 is communicated with the mixing hole 8 through the fourth auxiliary channel, and the waste liquid hole 4 is communicated with the test hole 9 through the fifth auxiliary channel. Since the functional units are four groups, the number of the respective holes is also four. For compactness and space saving, four sets of functional units are arranged in a cross around the circular main channel 1, and the structure shown in fig. 1 is that the central connecting lines of the serum collecting holes 2, the second auxiliary channels, the mixing holes 8, the third auxiliary channels and the testing holes 9 in the two opposite sets of functional units penetrate through the common center of the base body a and the main channel 1. The dilution liquid hole 3, the fourth auxiliary channel, the waste liquid hole 4 and the fifth auxiliary channel in each group of functional units rotate anticlockwise in a top view.
The main channel 1, the sample well 5, the blood cell collection well 7 and each of the functional units described above extend through the substrate a, facilitating both the flow of liquid and processing. In the same way, in consideration of fluidity and processing, each hole, the sample hole 5 and the blood cell collecting hole 7 in the functional unit are all round holes, and the diameters of the holes can be equal or unequal so as to meet various functional requirements.
For convenience of testing after the whole blood separation microfluidic chip is mounted to the detection device, each test well 9 is close to the edge of the substrate a as shown in fig. 1. In order to facilitate the inflow and outflow of the mixed solution and the reaction with the antibody serving as the detection probe, as shown in fig. 2, the hole wall 11 of the test hole 9 is provided with a sample inlet 10 and a liquid outlet 15 which are opposite to each other, the sample inlet 10 is close to the top of the test hole 9, and the liquid outlet 15 is close to the bottom of the test hole 9.
Example two
On the basis of the first embodiment, in the present embodiment, as shown in fig. 1, a whole blood separation microfluidic chip is provided with a positioning hole 6 on a substrate a, and the positioning hole 6 is located at the center of the substrate a and penetrates through the substrate a. In the embodiment, the positioning holes 6 are diamond holes, the positioning holes 6 are convenient for quick positioning when the microfluidic chip is mounted on the centrifuge, and the microfluidic chip is prevented from shifting in the centrifugation process, so that the microfluidic chip is ensured to shake stably in the whole centrifugation process.
In the above two embodiments, the number of the dilution holes 3 in each of the functional units is one, and the number of the dilution holes 3 in each of the functional units may be two, three or more depending on the concentration of the analyte.
Example III
As shown in fig. 2, on the basis of the second embodiment, the whole blood separation microfluidic chip further includes a centrifuge, the substrate a is connected to the centrifuge, and the bottom of the test well 9 is provided with a transparent substrate 12 for placing the detection probes 13 and the nanoparticles 14. In this embodiment, the transparent substrate 12 is a glass sheet, which is resistant to high temperatures, corrosion, heat and low in cost. The transparent substrate 12 may be a quartz sheet, a transparent ceramic sheet, a transparent polymer sheet, or the like, in addition to a glass sheet. The nanoparticle 14 is made of any one of pure gold, pure silver and pure platinum, or an alloy containing at least one of gold, silver and platinum.
In this embodiment, the centrifuge is a multistage variable speed centrifuge device. The centrifugal machine can adjust different rotating speeds at different stages of whole blood immunodetection so as to meet the processing requirements of detection samples at different stages.
Example IV
As shown in fig. 3, a whole blood separation microfluidic chip includes a centrifuge 20, and a substrate a connected to the centrifuge 20. As shown in fig. 2, a test well 9 penetrating the substrate is provided on the substrate, and a transparent substrate 12 for placing a detection probe 13 and a nanoparticle 14 is provided on the bottom 9 of the test well. In this embodiment, the transparent substrate 12 is a glass sheet, which is resistant to high temperatures, corrosion, heat and low in cost. The transparent substrate 12 may be a quartz sheet, a transparent ceramic sheet, a transparent polymer sheet, or the like, in addition to a glass sheet. The nanoparticle 14 is made of any one of pure gold, pure silver and pure platinum, or an alloy containing at least one of gold, silver and platinum.
Example five
As shown in fig. 3, a detection device includes a microfluidic chip, where the microfluidic chip is a whole blood separation microfluidic chip described in the third embodiment; one end of a test hole 9 in the whole blood separation microfluidic chip is provided with a parallel light component, and the other end is provided with a spectrum analysis component.
In this embodiment, the parallel light assembly includes a light source 31, an incoming optical fiber 32 and an incoming collimator 33, the light source 31 is connected with the incoming collimator 33 through the incoming optical fiber 32, the incoming collimator 33 is installed at one end of the test hole 9 far away from the spectrum analysis assembly, and the emission direction of the incoming collimator 33 is aligned with the test hole 9. The light source 31 is for emitting continuous visible white light in a wavelength range of 3333nm to 3330nm, and the light source 31 is a halogen lamp. The incoming fiber 32 serves as a carrier for light transmission, enabling light to be transmitted from the light source 31 to the incoming collimator 33. The incoming collimator 33 is used to prepare the parallel light rays and emit the parallel light rays to the test well 9.
In this embodiment, the spectrum analysis component includes a spectrometer 41, an outgoing optical fiber 42 and an outgoing collimator 43, where the spectrometer 41 is connected to the outgoing collimator 43 through the outgoing optical fiber 42, and the outgoing collimator 43 is installed at an end of the test hole 9 far away from the parallel light component, and the receiving direction of the outgoing collimator 43 is aligned with the test hole 9. The outgoing collimator 43 is used to receive the light after passing through the test aperture 9. The outgoing optical fiber 42 serves as a carrier for light conduction, enabling light to be conducted from the outgoing collimator 43 to the spectrometer 41. The spectrometer 41 analyzes the output signal of the outgoing optical fiber 42 to form a spectrum analysis result, and provides data support for subsequent body health analysis. The whole blood immunodetection device adopts the common optical fiber as the conducting medium, does not need complex optical path design, ensures that the structure of the device is more compact, is beneficial to reducing the whole volume of the device, and is suitable for families, communities and hospitals.
When the parallel light assembly emits light, the incoming collimator 33 is aligned with the test hole 9, the outgoing collimator 43 is aligned with the test hole 4, and the axes of the incoming collimator 33 and the outgoing collimator 43 are positioned on the same straight line with the axis of the test hole 9. In this embodiment, the incoming collimator 33 is located directly above the test hole 9 and the outgoing collimator 43 is located directly below the test hole 9. In other embodiments, the incoming collimator 33 is located directly below the test well 9 and the outgoing collimator 43 is located directly above the test well 4.
In this embodiment, the whole blood immunodetection apparatus further includes a human-computer interaction device 50, and the human-computer interaction device 50 is electrically connected with the light source 31 and the spectrometer 41, respectively. The human-computer interaction device 50 is a touch display screen for controlling the parallel light assembly, the spectrum analysis assembly and the centrifuge 20 and displaying the spectrum analysis result. The man-machine interaction device 50 comprises a Bluetooth device and a WIFI transceiver device and is used for exchanging data with external electronic equipment. Specifically, the output signal from the outgoing optical fiber 42 is analyzed by the human-machine interaction device 50 to form a spectral analysis result, or further analyzed by an external electronic device to form a more perfect spectral analysis result.
Example six
The whole blood detection method comprises the following steps:
1-3 drops of blood are dripped into a sample hole 5 of the whole blood separation microfluidic chip, the whole blood separation microfluidic chip is placed into a centrifugal machine 20, and the chip is fixed through a positioning hole 6 to prevent offset. Starting a centrifugal machine 20 to perform centrifugation, and separating blood, wherein blood cells enter a blood cell collecting hole 7, and serum enters a serum collecting hole 2 and flows into a mixing hole 8 under the action of centrifugal force;
the diluent in the diluent hole 3 enters the mixing hole 8 to be fully mixed with the serum, the mixed liquid is driven by centrifugal force to flow into the test hole 9, and the protein to be tested in the serum is specifically combined with the detection probe 13;
the mixed liquid in the test hole 9 flows into the waste liquid hole 4 under the drive of centrifugal force;
absorbance test was performed on test well 9.
After the test holes 9 in one group of functional units are tested, the rotary chip tests the test holes 9 in the other group of functional units, and so on until the test holes 9 in each functional unit are tested.
The whole blood separation micro-fluidic chip in the embodiment has the advantages of simple structure, convenient manufacture, suitability for market popularization and capability of realizing the rapid detection of whole blood. In the detection device and the whole blood detection method in the embodiment, the LSPR effect of the nano particles is utilized in the test process, so that the detection is pollution-free, real-time and high-sensitivity, and the concentration of the object to be detected is judged by detecting the absorbance change condition.
It is to be understood that the above examples of the present invention are provided for clarity of illustration only and are not limiting of the embodiments of the present invention. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. Any modification, equivalent replacement, improvement, etc. which come within the spirit and principles of the invention are desired to be protected by the following claims.
Claims (7)
1. The whole blood separation micro-fluidic chip comprises a centrifugal machine and a matrix connected with the centrifugal machine, and is characterized in that: the matrix is provided with a main channel, at least one group of functional units, and a sample hole and a blood cell collecting hole which are communicated with each other through the main channel;
the functional unit includes:
the serum collecting hole, the mixing hole and the testing hole are communicated in sequence, and the serum collecting hole is communicated with the main channel;
a diluent hole in communication with the mixing hole;
a waste well in communication with the test well;
the test hole penetrates through the matrix, and a transparent substrate for placing the detection probe and the nano particles is arranged at the bottom of the test hole;
the test hole is close to the edge of the substrate;
the hole wall of the test hole is provided with a sample inlet and a liquid outlet which are opposite, the sample inlet is close to the top of the test hole, and the liquid outlet is close to the bottom of the test hole.
2. The whole blood separation microfluidic chip according to claim 1, wherein: and the base body is provided with a positioning hole.
3. The whole blood separation microfluidic chip according to claim 1, wherein: the main channel is annular.
4. A detection device comprising a microfluidic chip, characterized in that: the microfluidic chip is the whole blood separation microfluidic chip according to any one of claims 1 to 3; one end of a test hole in the whole blood separation microfluidic chip is provided with a parallel light component, and the other end is provided with a spectrum analysis component.
5. The detection apparatus according to claim 4, wherein: the parallel light assembly comprises a light source, an incoming optical fiber and an incoming collimator, wherein the light source is connected with the incoming collimator through the incoming optical fiber, the incoming collimator is arranged at one end, far away from the spectrum analysis assembly, of the test hole, and the emission direction of the incoming collimator faces the test hole.
6. The detection apparatus according to claim 4, wherein: the spectrum analysis assembly comprises a spectrometer, an outgoing optical fiber and an outgoing collimator, wherein the spectrometer is connected with the outgoing collimator through the outgoing optical fiber, the outgoing collimator is installed at one end, far away from the parallel light assembly, of the test hole, and the receiving direction of the outgoing collimator faces the test hole.
7. A whole blood detection method, characterized by being applied to the detection device according to any one of claims 4 to 6, comprising the steps of:
dripping blood into a sample hole of a whole blood separation microfluidic chip, connecting the microfluidic chip with a centrifuge, starting the centrifuge, enabling blood cells to enter a blood cell collecting hole, enabling serum to enter a serum collecting hole and flowing into a mixing hole;
the diluent in the diluent hole enters the mixing hole to be mixed with the serum, the mixed liquid flows into the testing hole, and the protein to be tested in the serum is specifically combined with the detection probe;
the mixed liquid in the test hole flows into the waste liquid hole;
absorbance testing was performed on the test wells.
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| CN110090675B (en) * | 2019-05-15 | 2021-12-10 | 京东方科技集团股份有限公司 | Micro-fluidic chip, detection method thereof and micro total analysis system |
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