WO2019205780A1 - 一种微流控芯片及具有该微流控芯片的分析仪器 - Google Patents

一种微流控芯片及具有该微流控芯片的分析仪器 Download PDF

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Publication number
WO2019205780A1
WO2019205780A1 PCT/CN2019/075460 CN2019075460W WO2019205780A1 WO 2019205780 A1 WO2019205780 A1 WO 2019205780A1 CN 2019075460 W CN2019075460 W CN 2019075460W WO 2019205780 A1 WO2019205780 A1 WO 2019205780A1
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Prior art keywords
liquid
chamber
inlet
microfluidic chip
reagent
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PCT/CN2019/075460
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English (en)
French (fr)
Inventor
蒙玄
万惠芳
胡海升
李文美
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广州万孚生物技术股份有限公司
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Application filed by 广州万孚生物技术股份有限公司 filed Critical 广州万孚生物技术股份有限公司
Priority to US17/051,172 priority Critical patent/US20210123903A1/en
Priority to EP19792300.6A priority patent/EP3779435A4/en
Publication of WO2019205780A1 publication Critical patent/WO2019205780A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • B01L2200/146—Employing pressure sensors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N2035/00178—Special arrangements of analysers
    • G01N2035/00237—Handling microquantities of analyte, e.g. microvalves, capillary networks

Definitions

  • the present invention relates to the field of medical devices, and in particular to a microfluidic chip and an analytical instrument having the microfluidic chip.
  • IPD In Vitro Diagnosis
  • samples blood, body fluids, tissues, etc.
  • the instruments and reagents are required for the detection process.
  • An in vitro diagnostic system The system for in vitro diagnosis is roughly divided into two types; one is represented by the testing center laboratory, which has modular, automated, and pipelined sample testing, which also has high throughput, high efficiency, and high sensitivity. Advantages, but the entire system is also expensive, the volume is large, and the defects of professional operation are required. It is mainly used in large hospitals. The other is represented by point-of-care testing (POCT). Its system has integrated, miniaturized, sample testing anytime and anywhere, which also has the advantages of affordable price, simple operation and timely report. However, its test results have the disadvantages of sensitivity and low stability compared with the central laboratory.
  • POCT point-of-care testing
  • Microfluidic technology has been applied to in vitro diagnostic products at home and abroad.
  • Microfluidics is an interdisciplinary subject that controls microfluidics on a chip with microchannels, involving biological, chemical, fluid physics, electronics, optics, mechanical engineering, and more.
  • Microfluidic devices are commonly referred to as microfluidic chips and are also known as Lab on a Chips.
  • the basic operations of sample preparation, reaction, separation, and detection of biological, chemical, and medical analysis processes are usually concentrated on one chip to complete a system function.
  • the existing microfluidic chips are mainly qualitative detection, and the microfluidic chips for quantitative detection are few, and the existing quantitative microfluidic chips are complicated to prepare and have low production efficiency, such as the Chinese patent with the publication number “CN105214744”.
  • the application discloses "a magnetic particle chemiluminescence microfluidic chip", the microfluidic chip comprising a top plate and a bottom plate, wherein the top plate comprises an air pump, a sample filling port, a sample filling area, a labeled ligand storage pool and a sample a mixing zone; the bottom plate comprises a filtering zone, a magnetic particle coating zone, a cleaning zone, a detection zone, a cleaning liquid storage pool, a luminescent base liquid storage tank, and a liquid release channel; the top plate and the bottom plate both comprise a liquid sensing device, To determine the flow state of the liquid in the microfluidic chip and whether or not to mix air bubbles, etc., the chip in this patent adopts a multi-layer structure, and
  • the error is even larger. Compared with the microfluidic chip, the amount required is several tens of microliters. Therefore, the quantitative accuracy of the receiving bag cannot meet the requirements, and the quantitative accuracy is poor, which affects the detection result. At the same time, the bag needs to be built in. In the chip, the production difficulty of the chip is increased.
  • the present invention provides a microfluidic chip, which can realize quantitative detection, has a simple structure, and reduces the difficulty in manufacturing a chip.
  • a microfluidic chip comprising a chip body, an injection port disposed on the chip body, a liquid driving force inlet, a main fluid channel and a plurality of functional chambers;
  • the main fluid passage communicates with the plurality of functional chambers, the inlet and the liquid driving force inlet are respectively in communication with the main fluid passage, and the liquid driving force inlet is used for connecting a liquid driving device to drive the liquid Flowing into or out of the functional chamber;
  • At least one of the plurality of functional chambers is a liquid dosing chamber; the liquid dosing chamber has a predetermined volume, and a liquid identification site is provided at a liquid outlet of the liquid dosing chamber, and the liquid to be quantified Flowing from the liquid inlet of the liquid metering chamber into the liquid metering chamber, filling the liquid metering chamber and reaching the liquid outlet.
  • the liquid metering chamber includes a reagent metering chamber, the liquid inlet of the reagent metering chamber is in communication with one end of the reagent branch channel, and the other end of the reagent branch channel is in communication with the reagent inlet.
  • a liquid identification site is also provided at the inlet of the liquid metering chamber.
  • the liquid drive is a plunger pump.
  • the liquid dosing chamber further includes a sample quantification chamber through which the liquid sample flows into the sample dosing chamber for quantification; the sample dosing chamber is located in the reagent dosing chamber Upstream of the room;
  • the microfluidic chip is further provided with an air inlet and an air branch channel communicating therewith, one end of the air branch channel is connected to the air inlet, and the other end is connected to the sample quantification chamber and the inlet port.
  • the intermediate fluid passages are in communication, and the other end of the air branch passages is in communication with the main fluid passages adjacent to the sample metering chamber.
  • the functional chamber includes a detection chamber having a predetermined volume, and a liquid identification site is provided at a liquid outlet of the detection chamber, the liquid to be detected
  • the inlet of the detection chamber flows into the detection chamber, and fills the detection chamber to reach the liquid outlet.
  • a liquid identification site is also provided at the inlet of the detection chamber.
  • the liquid identification site is for positioning a liquid identification device;
  • the liquid identification device comprises a light source generation module and a photoelectric sensor;
  • the liquid recognition site includes an upper site for positioning the light source generating module and a lower site for positioning the photosensor, and the upper site and the lower site are respectively disposed outside the chip body.
  • the positions of the upper point and the lower point correspond to corresponding liquid outlets or liquid inlets, so that the positioned light source generating module, corresponding liquid outlet or liquid inlet, and the photoelectric sensor are vertical lines Layout in sequence.
  • the liquid metering chamber is a chamber of hexagonal configuration.
  • the liquid inlet chamber has a width of 0.3-3 mm and a height of 0.3-3 mm; the liquid metering chamber has a width of 0.3-3 mm and a height of 0.3. -3mm; or
  • the surface of the liquid metering chamber is a surface formed by modification of a hydrophilic surface;
  • the liquid inlet of the liquid metering chamber has a width of 0.3-5 mm and a height of 0.3-3 mm;
  • the outlet has a width of 0.3-5 mm and a height of 0.3-3 mm; or
  • the surface of the liquid metering chamber is a surface formed by a hydrophobic surface modification, the liquid metering chamber has a width of 0.3-2 mm and a height of 0.3-3 mm; the liquid metering chamber The outlet has a width of 0.3-2 mm and a height of 0.3-3 mm.
  • the chip body includes a top plate and a bottom plate; the top plate is laminatedly connected to the bottom plate, and the main fluid passage and the plurality of functional chambers are disposed at a connection between the top plate and the bottom plate room.
  • the bottom plate is a smooth plate
  • the top plate is provided with micropores, microchannels or microcavities to form the inlet, the liquid driving force inlet, and the main fluid channel with the bottom plate. Or function chamber.
  • the inlet and the liquid driving force inlet are respectively disposed at both ends of the main fluid passage.
  • the present invention also provides an analytical instrument having a microfluidic chip, comprising an instrument frame, at least one reagent storage pool, a liquid driving device, a detecting device, and the microfluidic chip described above;
  • the microfluidic chip is mounted in the instrument frame;
  • the liquid driving device is connected to a liquid driving force inlet of the microfluidic chip;
  • the reagent storage pool and the corresponding reagent inlet are communicably connected;
  • the detecting device It is used for receiving and processing a detection signal sent by the microfluidic chip.
  • the liquid drive device is a plunger pump; each of the reagent storage tanks is provided with an opening in communication with outside air.
  • the present invention has the following beneficial effects:
  • the microfluidic chip provided by the invention realizes the quantification of the liquid by a specific liquid quantification chamber combined with the liquid driving device, and the liquid quantification chamber of the present invention is compared with the prior art to realize the quantification by pressing the reagent package embedded in the chip.
  • the chamber can improve the accuracy of the quantification; and the reagent can be externally placed on the chip, and the multi-layer chip combination and the reagent package are embedded in the chip in the prior art, which reduces the manufacturing process difficulty of the chip and improves the detection accuracy.
  • the chip body of the microfluidic chip of the invention may comprise a top plate and a bottom plate which are arranged in a stack, and the top plate which can be arranged by the processed structure, the bottom plate is only a smooth flat plate, which can further reduce the manufacturing process difficulty of the chip and improve the production. effectiveness.
  • FIG. 1 is a schematic structural view of an embodiment of a microfluidic chip provided by the present invention.
  • Figure 2 is a schematic cross-sectional view of a liquid identification device provided by the present invention.
  • FIG. 3 is a structural view showing a sensor arrangement of an embodiment of a microfluidic chip provided by the present invention.
  • FIG. 4 is a schematic cross-sectional view showing a position where a magnet is disposed in use of the microfluidic chip provided by the present invention
  • Figure 5 is a schematic structural view of an embodiment of a liquid driving device provided by the present invention.
  • the embodiment provides a microfluidic chip, the microfluidic chip includes a chip body, and an inlet, a liquid driving force inlet, a main fluid channel and a plurality of functional chambers disposed on the chip body; Detailed instructions are given.
  • the primary fluid channel communicates with a plurality of functional chambers to direct the flow of fluid between the functional chambers.
  • the functional chamber has at least a function of accommodating.
  • the functional chamber has other functions in addition to the accommodating function.
  • Other functions may be implemented in the functional chamber, or may be combined with the functional chamber.
  • the necessary external components (these necessary components can be fixed outside the chip, without being placed inside the chip or on the surface).
  • the inlet and the liquid driving force inlet are respectively in communication with the main fluid passage, the liquid driving force inlet is for connecting the liquid driving device to drive the liquid to flow into or out of the functional chamber, and the inlet is for introducing the liquid sample In the main fluid channel, the liquid sample enters each functional chamber through the main fluid channel.
  • At least one of the plurality of functional chambers is a liquid dosing chamber; the liquid dosing chamber has a predetermined volume, and a liquid identification site is disposed at a liquid outlet of the liquid dosing chamber, The metered liquid flows from the liquid inlet of the liquid metering chamber into the liquid metering chamber, and fills the liquid metering chamber to reach the liquid outlet.
  • the liquid identification site is used to position or fix the liquid identification device, and the liquid identification device is used to identify the liquid.
  • the liquid identification device can recognize the liquid and provide the liquid arrival signal; thus when the liquid reaches the liquid discharge At the mouth, the liquid identification device can provide a liquid arrival signal indicating that the liquid has filled the liquid metering chamber, at which point the liquid drive device is controlled to stop driving the liquid to achieve metering of the liquid in the liquid metering chamber.
  • the microfluidic chip provided by the embodiment realizes the quantification of the liquid by a specific liquid quantification chamber combined with the liquid driving device, and the liquid quantification of the present invention is compared with the prior art to realize the quantification by pressing the reagent package embedded in the chip.
  • the chamber increases the accuracy of the quantification.
  • the reagent can be externally placed on the chip, and the multi-layer chip combination and the reagent package are embedded in the chip in the prior art, which reduces the manufacturing process difficulty of the chip and improves the detection accuracy.
  • the main fluid channel and the plurality of functional chambers may be formed inside the chip body by laser processing, model injection molding, or the like, or may be processed on the top plate or the bottom plate by being provided as a separate top plate and bottom plate.
  • the chip body includes a top plate and a bottom plate; the top plate is laminated with the bottom plate; and the connection between the top plate and the bottom plate is provided
  • the main fluid channel and the plurality of functional chambers more preferably, the bottom plate is a smooth plate, and the top plate is provided with micropores, microchannels or microcavities to cooperate with the bottom plate to form an inlet, a liquid driving force inlet, a main fluid
  • the channel or the plurality of functional chambers, such a microfluidic chip is more convenient to prepare, reduces the difficulty of the production process, and only needs to process the specific structure required on the top plate, thereby improving the production efficiency.
  • the bottom plate is a smooth flat plate
  • the top plate is provided with a plurality of microchannels to form a main fluid passage with the bottom plate
  • the top plate is provided with a plurality of microcavities to be combined with the bottom plate to form a a plurality of functional chambers
  • the top plate being provided with a plurality of holes for combining with the bottom plate to form the inlet or the liquid driving force inlet; for facilitating the injection, the size of the inlet is generally larger than the size of the other inlets .
  • the number of liquid dosing chambers, the type of liquid they are metered in eg, liquid samples, reagents, sample processing reagents, etc.
  • the location of the set, and the type of other functional chambers can be selected based on actual needs.
  • the liquid quantification chamber comprises a reagent quantification chamber
  • the liquid inlet of the reagent quantification chamber is in communication with one end of the reagent branch channel
  • the other end of the reagent branch channel is in communication with the reagent inlet
  • the reagent enters the reagent through the reagent inlet and the reagent branch channel.
  • the quantification chamber is quantified.
  • the amount of liquid sample ie, the liquid to be tested flowing from the inlet
  • reagents eg, reagents, sample processing reagents, etc.
  • the reagent needs to be quantified in the chip.
  • the liquid sample can optionally be quantified outside of the microfluidic chip.
  • the reagent quantification chamber is used for quantification of the reagent.
  • the "liquid metering chamber includes a reagent metering chamber” as used herein shall be understood to mean that the liquid metering chamber should have at least a type of reagent metering chamber for metering reagents, and of course may further comprise a liquid for quantifying other liquid types.
  • a quantification chamber such as a sample quantification chamber for quantifying a liquid sample, and the like.
  • the number of reagent quantification chambers may be one, two or more, and may be selected according to the actual needs of the microfluidic chip.
  • the microfluidic chip is a chemiluminescent microfluidic chip
  • at least one reagent quantification chamber should be provided for quantifying the substrate illuminating liquid, and the remaining reaction essential substances such as the enzyme standard primary antibody.
  • the magnetic standard secondary antibody can be respectively embedded in the two functional chambers of the enzyme standard primary anti-embedding chamber and the magnetic standard secondary anti-embedding chamber; preferably, the magnetic standard secondary anti-embedding chamber is a reagent quantitative chamber
  • the magnetic secondary antibody is not only embedded in the magnetic secondary antibody, but also used to quantify the substrate luminescent liquid; more preferably, the magnetic secondary anti-embedding chamber is also used for magnetic bead cleaning.
  • the reagent inlet and the reagent storage pool are connected to each other through a valve, and the reagent storage pool is provided with an opening communicating with the outside air, and the reagent storage pool is provided with an opening to facilitate the liquid driving device to drain the liquid therein into the chip.
  • the reagent storage pool is disposed outside the microfluidic chip, and in use, the reagent storage pool is installed at the reagent inlet to introduce the reagent into the chip.
  • a liquid identification site is also provided at the inlet of the liquid metering chamber.
  • the liquid identification site can also be used to position or immobilize a liquid identification device that is arranged to facilitate monitoring and control of the flow of liquid within the chip, which also enables mixing between two metered liquids, such as between a liquid sample and a reagent. the mix of.
  • the microfluidic chip of the invention needs to simultaneously realize the quantification of the liquid and the contact of the two liquids, which requires
  • a metered liquid stays at a predetermined position, and another liquid preferably flows from the predetermined position into the liquid metering chamber to achieve quantification in the liquid metering chamber, and the optimal choice of the predetermined position is the liquid metering chamber
  • a liquid identification point is provided at the liquid inlet to position the liquid identification device, and the liquid identification device can provide a stop indication signal of one of the liquids and an infusion signal of the other liquid in the liquid quantitative chamber
  • the liquid identification device can provide a stop indication signal of one of the liquids and an infusion signal of the other liquid in the liquid quantitative chamber
  • the reagent inlet and the reagent storage pool can be connected to each other through a valve, and the reagent storage pool is provided with an opening communicating with the outside air.
  • the liquid sample (in order to facilitate accurate mixing or reaction with the quantitative reagent, the liquid sample is preferably a predetermined amount of liquid sample) flows from the inlet through the main fluid channel to the inlet of the reagent dosing chamber under the action of the liquid driving device.
  • the liquid identification device positioned at the liquid identification site of the reagent quantification chamber acquires a signal, and controls the liquid drive device to stop driving.
  • the liquid sample stops flowing, and the front end of the liquid sample stays at the liquid inlet; then, Close the air inlet, open the valve between the reagent inlet and the reagent storage tank, and the reagent enters the reagent quantitative chamber from the reagent inlet through the reagent inlet channel and the reagent inlet chamber through the liquid driving device, and flows to the reagent quantitative solution.
  • the reagent is filled with the reagent quantification chamber, at which time the valve between the reagent inlet and the reagent storage tank is closed, the air inlet is opened, and the reagent and liquid sample of the reagent quantification chamber are continued under the action of the liquid drive device. Flow, and can realize positive reverse motion under the action of positive pressure and negative pressure of liquid driving device Mixing and / or reaction.
  • one end of the reagent branch channel passes through the main fluid channel and the reagent dosing chamber
  • the liquid port is connected, and the communication branch channel and the main fluid channel are adjacent to the liquid inlet of the reagent quantitative chamber, so that the quantitative is within a controllable range, and the quantitative error is reduced, for example, the reagent branch channel
  • the distance from the inlet of the main fluid passage from the inlet of the reagent metering chamber is 0.5 to 10 mm (preferably 0.5 to 2 mm).
  • the inlet and the liquid driving force inlet are respectively disposed at both ends of the main fluid passage.
  • the functional chamber comprises a detection chamber, the detection chamber has a predetermined volume, and a liquid identification site is arranged at the liquid outlet of the detection chamber, and the liquid to be detected flows into the detection chamber, and the detection is full. After the chamber reaches the liquid outlet.
  • the liquid identification site provided at the liquid outlet of the detection chamber can be used for positioning or fixing the liquid identification device.
  • the liquid identification device sends a signal, and the liquid drive device controls The detected liquid stops flowing and can be detected at this time.
  • a liquid identification site is also provided at the liquid inlet of the detection chamber.
  • the liquid quantification chamber further comprises a sample quantification chamber, wherein the liquid sample flows into the sample quantification chamber through the injection port for quantification; the sample quantification chamber is located upstream of the reagent quantification chamber; and the microfluidic chip is further provided with An air inlet and an air branch passage communicating therewith, one end of the air branch passage communicating with the air inlet and the other end communicating with a main fluid passage between the sample metering chamber and the inlet, the air The communication between the other end of the branch channel and the main fluid channel is adjacent to the sample dosing chamber; herein "adjacent” is generally understood to mean “0.5 to 10 mm (preferably 0.5-2 mm) from the inlet of the sample dosing chamber. )".
  • the air inlet and the air pipe outside the chip can be connected to each other through the valve to control the air to enter the inside of the chip.
  • the liquid sample flows into the sample quantification chamber from the inlet of the sample quantification chamber through the inlet through the inlet of the liquid driving device, and when the liquid sample flows to the liquid outlet of the sample quantification chamber, the sample dosing chamber is filled.
  • the liquid identification device located at the liquid identification point of the liquid outlet emits an indication signal to control the opening of the air inlet, and the driving force required for the flow of the liquid sample is small, and the driving force required for the flow of the liquid sample is small.
  • the liquid sample stays in the communication between the air branch channel and the main fluid channel and no longer flows into the sample dosing chamber, so that the liquid sample can be quantified in the sample dosing chamber.
  • the quantified liquid sample can continue to flow to the inlet of the reagent quantification chamber under the action of the liquid driving device.
  • the reagent quantification chamber completes the quantification of the reagent (the process is as described above)
  • the liquid sample and the reagent are driven in the liquid.
  • the mixing and/or reaction of the device is achieved by alternating positive and negative pressures.
  • the microfluidic chip in this embodiment can facilitate the quantification of the liquid sample by setting the sample quantification chamber, and does not need to be separately quantified outside the chip, so that the chip is more convenient to use.
  • the liquid sample is whole blood
  • a whole blood filtering chamber is disposed between the inlet and the sample quantification chamber
  • a whole blood filter membrane is disposed in the whole blood filter chamber
  • whole blood is a common test sample.
  • the chamber is convenient for detection and use, and compared with the method of first quantifying whole blood and then separating the whole blood
  • a whole blood filtering chamber is arranged between the inlet and the sample quantification chamber, and the sample quantification chamber can be passed. The amount of serum or plasma is directly quantified in the room, and the measurement result is more accurate.
  • the whole blood filter membrane may be made of glass fiber, cotton linters, polyester fibers, fibers or blended fibers; optionally, the whole blood filter mat has a thickness of 0.2-2.5 mm; the whole blood
  • the filtration filter has an adsorption rate of 4-150 s/4 cm and a water absorption of 30-250 mg/cm 2 .
  • the outlet of the whole blood filtration zone is a triangular outlet;
  • the whole blood filtration area is 30-300 mm 2 , the width is 2-20 mm, the length is 5-25 mm, the depth is 0.3-3 mm, and the front end is triangular.
  • the angle is 15-160 °C.
  • the embodiment provides a chemiluminescent microfluidic chip, which comprises a chip body, an inlet 2 disposed on the chip body, a liquid driving force inlet 13, and a substrate illuminating liquid inlet.
  • a chemiluminescent microfluidic chip which comprises a chip body, an inlet 2 disposed on the chip body, a liquid driving force inlet 13, and a substrate illuminating liquid inlet.
  • the primary fluid channel communicates with a plurality of functional chambers to direct the flow of fluid between the functional chambers.
  • the functional chamber includes an enzyme-labeled anti-embedding region 5, a magnetic-labeled secondary antibody-embedded region 7, and a chemiluminescent detection region 9 that are sequentially connected through the main fluid channel.
  • the enzyme-labeled primary anti-embedding zone 5 is embedded with the enzyme-labeled primary antibody;
  • the magnetic-labeled secondary-resistance zone 7 is embedded with the magnetic-labeled secondary antibody;
  • the magnetic-labeled secondary antibody-embedded zone 7 is the liquid quantitative chamber;
  • the chamber is used to quantify the liquid, and after the liquid to be quantified (for example, the substrate illuminating liquid) enters the liquid quantification chamber, the quantification can be achieved in the liquid quantification chamber (ie, the desired amount of liquid is obtained) to be used with the quantitative liquid sample or Other reagents react to achieve quantitative detection.
  • the liquid metering chamber has a predetermined volume
  • a liquid identification site is disposed at the liquid outlet of the liquid metering chamber, and the liquid to be quantitatively flows from the liquid inlet of the liquid metering chamber into the liquid metering chamber.
  • the chamber is filled with the liquid quantification chamber and reaches the liquid outlet; the liquid identification site is used to locate or fix the liquid identification device, and the liquid identification device is used to identify the liquid.
  • the liquid identification device can provide a liquid arrival signal indicating that the liquid has filled the liquid metering chamber, and at this time, controlling the liquid driving device to stop driving the liquid, the liquid can be quantified in the liquid quantitative chamber.
  • the chemiluminescent microfluidic chip achieves the quantification of the liquid by a specific liquid dosing chamber combined with a liquid driving device, which can improve the accuracy of the quantification.
  • the chemiluminescence detection zone 9 is used to house the chemiluminescent reaction product to complete the detection process in combination with an external detection device.
  • the inlet 2 and the liquid driving force inlet 13 are respectively in communication with the main fluid passage, the driving force inlet 13 is for connecting the liquid driving device to drive the liquid into or out of the functional chamber; the inlet 2 is for introducing the liquid sample into the main fluid passage The liquid sample enters each functional chamber through the main fluid channel.
  • one end of the substrate illuminating liquid branch channel 17 is in communication with the substrate luminescent liquid inlet 11, and the other end thereof is in communication with the liquid inlet of the magnetic standard secondary anti-embedding region 7, and the substrate illuminating liquid is illuminated by the substrate.
  • the liquid inlet 11, the substrate illuminating liquid branch channel 17 enters the magnetic standard secondary anti-embedding zone 7 for quantification.
  • One end of the cleaning liquid branch passage 18 communicates with the cleaning liquid inlet 12, and the other end thereof communicates with the liquid inlet of the magnetic standard secondary anti-embedding area 7, and the cleaning liquid enters the magnetic standard secondary antibody through the cleaning liquid inlet 12 and the cleaning liquid branch passage 18.
  • the embedding area 7 performs magnetic bead cleaning.
  • the substrate illuminating liquid inlet 11 and the cleaning liquid inlet 12 are respectively connected to the substrate illuminating liquid storage tank 22 and the cleaning liquid storage tank 23 through the valves V2 and V3, and the substrate is connected.
  • the illuminating liquid storage tank 22 and the cleaning liquid storage tank 23 are respectively provided with openings communicating with the outside air;
  • the liquid driving device is installed at the liquid driving force inlet 13 for driving the liquid flow in the chip;
  • the magnetic standard secondary anti-embedding area 7 Magnets (for example, magnets 25a, 25b) are fixed to the outside to fix the magnetic beads 26.
  • the magnetically labeled secondary anti-embedding zone is a liquid dosing chamber that can be used to quantify the substrate illuminating liquid and, optionally, can be further used to quantify the cleaning solution.
  • the working mode of the microfluidic chip of the embodiment is as follows: a predetermined amount of liquid sample (such as serum or plasma diluted by the diluent) flows from the inlet 2 through the main fluid channel under the action of the liquid driving device to The enzyme-labeled primary antibody-embedded region 5 is mixed with the enzyme-labeled primary antibody, and then the reaction solution reaches the magnetic-labeled secondary antibody-embedded region 7, and the magnetic hybrid secondary antibody is continuously mixed and reacted therein.
  • a predetermined amount of liquid sample such as serum or plasma diluted by the diluent
  • the reactants of the double-anti-sandwich structure are formed on the beads, the magnetic beads are adsorbed by the magnets, and the reactants are stabilized in the magnetic-labeled secondary anti-embedding zone 7 by the magnetic beads, and the remaining reaction liquids are under the action of the liquid driving device.
  • the liquid driving force inlet 13 discharges the chip; then, the air inflow port (such as the sample inlet) on the chip is closed, and the valve V3 between the cleaning liquid storage tank 23 and the cleaning liquid inlet 12 is opened, and the cleaning liquid is subjected to the action of the liquid driving device.
  • the cleaning liquid branch channel 18 enters the magnetic standard secondary anti-embedding area 7 to clean the magnetic beads therein, and when the magnetic standard secondary anti-embedding area 7 completes the quantification of the cleaning liquid, the cleaning liquid storage tank 23 can be closed and cleaned.
  • Valve V3 between liquid inlet 12, open The gas flows into the port, and the cleaned liquid is discharged from the chip through the liquid driving force inlet 13 under the action of the liquid driving device.
  • the cleaning may be repeated several times (the magnetic bead cleaning method is not limited to the method described herein, and may also pass For example, moving the magnet in the cleaning solution to achieve cleaning of the magnetic beads; then closing the air inflow port on the chip (such as the sample inlet), opening the valve V2 between the substrate luminescent liquid storage pool 22 and the substrate luminescent liquid inlet 11,
  • the substrate illuminating liquid enters the magnetic standard secondary anti-embedding area 7 through the substrate illuminating liquid branch channel 19 under the action of the liquid driving device, and when the magnetic standard secondary anti-embedding area 7 completes the quantification of the substrate luminescent liquid, the bottom is closed.
  • the valve V2 between the illuminating liquid storage tank 22 and the substrate illuminating liquid inlet 11 stops the driving action of the liquid driving device, and the substrate illuminating liquid no longer flows into the magnetic label secondary anti-embedding area 7, and opens the air inflow port on the chip ( For example, if the sample is imported, the substrate illuminating liquid after quantification of the magnetic standard secondary antibody is luminescent reaction with the reactant captured by the magnetic bead, and then the magnet is removed, and the reaction liquid in the magnetic standard secondary anti-embedding region 7 is in the liquid driving device. Chemiluminescent detection by the inflow region 9 is detected.
  • the above-mentioned chemiluminescent microfluidic chip has a compact structure, for example, the magnetic standard secondary anti-embedding region is not only used for embedding the magnetic secondary antibody, but also can be used as a liquid quantitative chamber for quantifying the substrate illuminating liquid without further setting of liquid quantification.
  • the chamber and the magnetic standard secondary anti-embedding area can further serve as a region for magnetic bead cleaning, without separately providing a magnetic bead cleaning area, which greatly saves the volume of the chip; at the same time, the reagent storage pool (such as the substrate illuminating liquid)
  • the storage pool, the cleaning liquid storage pool, etc. can be externally placed on the chip, and is embedded in the chip compared with the prior art reagent package, which reduces the manufacturing process difficulty of the chip and improves the detection accuracy.
  • the main fluid channel and the plurality of functional chambers may be formed inside the chip body by laser processing, model injection molding, or the like, or may be processed on the top plate or the bottom plate by being provided as a separate top plate and bottom plate.
  • the chip body comprises a top plate 1 and a bottom plate 20; the top plate 1 and the bottom plate 20 are laminatedly connected; the top plate 1 and the bottom plate 20 are The connection is provided with a main fluid channel and a plurality of functional chambers; more preferably, the bottom plate 20 is a smooth flat plate, and the top plate 20 is provided with micropores, microchannels or microcavities to cooperate with the bottom plate to form an inlet 2, a liquid driving force
  • the bottom plate 20 is a smooth flat plate
  • the top plate 1 is provided with a plurality of microchannels to form a main fluid passage with the bottom plate 20, and the top plate 1 is provided with a plurality of microcavities for combining with the bottom plate 20 to form a plurality of functions.
  • a chamber the top plate 1 is provided with a plurality of holes to combine with the bottom plate 20 to form an injection port 2, a liquid driving force inlet 13, a substrate illuminating liquid inlet 11 and a cleaning liquid inlet 12; for the purpose of facilitating the injection, the inlet 2
  • the size is usually larger than the size of the other inlets.
  • the chip body of the above-mentioned chemiluminescent microfluidic chip may include a top plate and a bottom plate which are arranged in a stack, and the top plate which can be disposed on the structure to be processed is only a smooth flat plate, which can further reduce the manufacturing process difficulty of the chip. Increase productivity.
  • a liquid identification site is also provided at the inlet of the liquid metering chamber.
  • the liquid identification site is arranged to facilitate monitoring and control of the flow of liquid in the chip and possible air bubbles, and also enables mixing between two quantitative liquids, such as liquid samples and reagents (eg, reagents, sample processing). Mixing between reagents, etc.).
  • the enzyme-labeled primary anti-embedding region 5 is also a liquid quantitative chamber, and the chip body is further provided with a diluent inlet 10 and a diluent branch channel 16; one end of the diluent branch channel 16 is connected to the diluent inlet 10, and One end is connected with the inlet port of the enzyme-labeled primary anti-embedding zone 5, and the sample diluent is quantified by the diluent inlet and the diluent branch channel into the enzyme-labeled primary anti-embedding zone 5.
  • a liquid identification site is respectively arranged at the liquid inlet and the liquid outlet of the enzyme-labeled anti-embedding zone 5, and the liquid to be quantitatively flows from the liquid inlet into the enzyme-labeled primary anti-embedding zone 5, filled with the enzyme After the first anti-embedding zone 5 is reached, the liquid outlet is reached.
  • the sample diluent can not only dilute the liquid sample (such as serum, plasma, etc.), but also reduce its concentration and viscosity.
  • the substance contained in it can also reduce the background value of the liquid sample, making the detection more accurate, and the sample diluent can be more
  • a good reconstituted enzyme is labeled as a primary antibody; in this technical solution, the enzyme-labeled primary anti-embedding region can be used to quantify the sample diluent without the need to quantify the sample diluent outside the chip, and the quantitative sample diluent can be in the enzyme label.
  • the mixing of the primary anti-embedding zone with the quantitative liquid sample saves manpower and makes the operation more convenient.
  • the diluent inlet 10 and the diluent storage tank 21 are connected to each other through the valve V1, and the diluent storage tank 21 is provided with an opening communicating with the outside air; a predetermined amount of the liquid sample (if diluted by the diluent) The serum or plasma) is flowed from the inlet 2 through the main fluid channel to the inlet of the enzyme-labeled anti-embedding zone 5 by the liquid driving device, and the air inlet (such as the sample inlet) on the chip is closed.
  • the sample diluent enters the enzyme-labeled primary anti-embedding zone 5 through the diluent branch channel 16 under the action of the liquid driving device, when it is filled with the enzyme-labeled primary antibody
  • the chemiluminescence detection zone 9 has a predetermined volume, and liquid identification sites are respectively disposed at the liquid inlet and the liquid outlet of the chemiluminescence detection zone 9, and the liquid to be detected passes through the chemiluminescence detection zone 9
  • the liquid port flows into the chemiluminescence detection zone 9, and is filled with the chemiluminescence detection zone 9 to reach the liquid outlet, and the volume of the chemiluminescence detection zone 9 is less than or equal to the volume of the magnetic resonance secondary embedding zone 7.
  • the liquid identification site provided at the liquid outlet of the chemiluminescence detection zone 9 can be used to locate or fix the liquid identification device, and the reaction liquid after the substrate luminescent liquid reacts with the reactant captured by the magnetic beads reaches the chemiluminescence detection zone.
  • the liquid identification device sends a signal, and the liquid drive device controls the reaction liquid to stop flowing, and the detection can be performed at this time.
  • the main fluid channel includes a first mixing channel 6 and a second mixing channel 8; the first mixing channel 6
  • the second mixing channel 8 is disposed between the magnetic standard secondary antibody embedding region 7 and the chemiluminescent detection region 9.
  • the injection port 2 and the liquid driving force inlet 13 are respectively disposed at both ends of the main fluid passage.
  • the chip main body is provided with a magnet fixing site at a position corresponding to the magnetic standard secondary anti-embedding area 7; further, since the cleaning of the magnetic beads can be performed on the magnetic label
  • the second anti-embedding area 7 is carried out.
  • a magnet fixing point for positioning the magnets 25a, 25b and two magnets 25a are disposed above and below the magnetic labeling secondary anti-embedding area 7.
  • 25b corresponds to the oblique diagonal arrangement of the magnetic-labeled secondary anti-embedding zone 7.
  • the liquid driving device is the plunger pump 19, and the description of the plunger pump in Embodiment 3 is applicable to the present embodiment.
  • the function chamber further includes a sample quantification chamber 4, the sample quantification chamber 4 is also a liquid quantification chamber, and the liquid sample flows into the sample quantification chamber 4 through the inlet port for quantification; the sample quantification chamber 4 is located at the enzyme label An upstream of the anti-embedding zone 5; the microfluidic chip is further provided with an air inlet 14 and an air branch channel 27 communicating therewith, one end of the air branch channel 27 is in communication with the air inlet 14, and the other end is connected to the sample dosing chamber 4 and The main fluid passage between the injection ports 2 communicates, and the other end of the air branch passage 27 communicates with the main fluid passage adjacent to the sample dosing chamber 4.
  • adjacent is generally understood to mean “1 to 10 mm from the inlet of the sample dosing chamber 4.”
  • the quantification of the liquid sample can be facilitated without additional quantification outside the chip, making the chip more convenient to use.
  • a liquid identification site is disposed at the liquid outlet of the sample quantification chamber 4, and the liquid to be quantified flows from the liquid inlet to the sample quantification chamber 4, and fills the sample quantification chamber 4 and reaches the liquid outlet. Further, a liquid identification site is also provided at the liquid inlet of the sample dosing chamber 4.
  • the air inlet and the air pipe outside the chip can be connected to each other through the valve to control the air to enter the inside of the chip.
  • the liquid sample flows into the sample quantification chamber from the inlet of the sample quantification chamber through the inlet through the inlet of the liquid driving device, and when the liquid sample flows to the liquid outlet of the sample quantification chamber, the sample dosing chamber is filled.
  • the liquid identification device located at the liquid identification point of the liquid outlet emits an indication signal to control the opening of the air inlet, and the driving pressure required for the flow of the air in the air branch passage is small, and the flow of the liquid sample is required to be driven.
  • the pressure is greater, so that the liquid sample stays in the communication between the air branch channel and the main fluid channel and no longer flows into the sample dosing chamber, thereby completing the quantification of the liquid sample in the sample dosing chamber.
  • the quantified liquid sample can continue to flow to the enzyme-labeled primary anti-embedding zone under the action of the liquid driving device.
  • the liquid sample is whole blood
  • a whole blood filtration zone 3 is disposed between the injection port 7 and the sample quantification chamber 4, and a whole blood filter membrane is disposed in the whole blood filtration zone 3; when the microfluidic chip is used In clinical diagnosis, whole blood is a common test sample. When testing, it is usually necessary to separate whole blood to separate the serum or plasma in the whole blood, and then react with the reagent; the whole blood filtration zone is set in the chip, which is convenient for detection and use.
  • a whole blood filtration zone is arranged between the inlet and the sample quantification chamber, and the amount of serum or plasma can be directly quantified through the sample quantification chamber, and the measurement is performed.
  • the whole blood filter membrane may be made of glass fiber, cotton linters, polyester fibers, fibers or blended fibers; optionally, the thickness of the whole blood filter mat is 0.2-2.5 mm; the adsorption speed of the whole blood filter mat It is 4-150 s/4 cm and has a water absorption of 30-250 mg/cm 2 .
  • Example 4 The description of the liquid dosing chamber in Example 4 is applicable to the description of the liquid dosing chamber described above (including the magnetic target secondary anti-embedding zone 7, the enzyme-labeled primary anti-embedding zone 5, and the sample quantification compartment 4). I will not repeat them here.
  • Embodiment 5 The description of the liquid identification site and the liquid identification device in Embodiment 5 is applicable to the description of the liquid recognition site and the liquid identification device described above, and details are not described herein again.
  • the other end of the substrate illuminating liquid branch channel 17 and the inlet port of the magnetic standard secondary anti-embedding area 7 are located on the main fluid channel of the liquid inlet of the magnetic standard secondary anti-embedding area 7;
  • "adjacent” herein is understood to mean “0.5 to 10 mm (preferably 0.5 to 2 mm) from the inlet of the magnetic secondary interference zone 7".
  • the cleaning liquid enters the magnetic standard secondary anti-embedding area 7 through the cleaning liquid inlet 12 and the cleaning liquid branch channel 18 for quantification; the other end of the cleaning liquid branch channel 18 and the liquid inlet of the magnetic standard secondary anti-embedding area 7
  • the communication is located on the main fluid passage adjacent to the liquid inlet; in one embodiment, "adjacent” is understood herein to mean “0.5 to 10 mm (preferably 0.5 to) from the inlet of the magnetic secondary interference zone 7. 2mm)".
  • the other end of the cleaning liquid branch channel 18 communicates with the liquid inlet of the magnetic standard secondary anti-blocking zone 7 at the other end of the substrate illuminating liquid branching channel 17 and the magnetic standard secondary anti-embedding zone 7 Downstream of the mouth, this prevents the substrate luminescent liquid from being diluted by the cleaning solution.
  • the other end of the diluent branch channel 16 and the inlet of the enzyme-labeled anti-embedding zone 5 are located on the main fluid channel adjacent to the liquid inlet of the enzyme-labeled primary anti-embedding zone 5;
  • "adjacent” herein is understood to mean “0.5 to 10 mm (preferably 0.5 to 2 mm) from the inlet of the enzyme-labeled primary anti-embedding zone 5".
  • the inlet 2 has a volume of 5 ul to 300 ul.
  • the liquid outlet of the whole blood filtration zone 3 is a triangular liquid outlet; the whole blood filtration zone 3 has an area of 30-300 mm 2 , a width of 2-20 mm, a length of 5-25 mm, and a depth of 0.3-3 mm.
  • the angle of the triangle is 15-160 °C.
  • the sample dosing chamber 4 has a volume of 1-50 ul.
  • the volume of the enzyme-labeled primary antibody-resistant region 5 is 5-50 ul.
  • the first mixing pipe 6 and the second mixing pipe 8 have a width of 200-2000 um, a length of 5 mm-40 mm, and a depth of 0.2-3 mm.
  • the magnetic secondary interference zone 7 has a volume of 10-200 ul.
  • the chemiluminescence detection zone 9 has a volume of 10-200 ul.
  • the method includes steps 101 to 110, and the steps are as follows:
  • Step 101 inserting a steel needle respectively connected to the diluent storage tank 21, the substrate illuminating liquid storage tank 22, the cleaning liquid storage tank 23, the plunger pump 19, and the air into the sealing pad 15 in the chip, wherein the steel needle is separately diluted
  • the liquid inlet 10, the substrate illuminating liquid inlet 11, the cleaning liquid inlet 12, the liquid driving force inlet 13, and the air inlet 14 are connected; the whole blood sample is added to the inlet 2, the electromagnetic valve V4 is opened, and the plunger pump 19 generates a negative
  • the whole blood sample is drawn into the whole blood filtration zone 3 by pressure suction.
  • Step 102 The serum after the completion of the filtration of the whole blood sample is taken into the sample quantification chamber 4, and the quantitative measurement of the serum is performed by the photoelectric sensors (a1, a2) provided on the inlet port and the liquid outlet of the sample quantification chamber 4.
  • the output voltage value of the sensor changes, giving the system an identification signal to determine the flow position of the liquid in the chip.
  • the sample passes through the photosensor a2, it is judged that the sample fills the sample dosing chamber 4, and the intrinsic volume of the region is the quantitative value of the sample.
  • Step 103 Block the injection port 2 and open the solenoid valve V5 so that the serum is inhaled into the enzyme-labeled primary antibody-resistant region 5.
  • Step 104 When the serum is detected by the photoelectric sensor (b1) provided on the inlet of the enzyme-labeled anti-embedding zone 5, the solenoid valve V5 is closed, and the solenoid valve V1 is opened, so that the external sample diluent enters from the solenoid valve V1. Enzyme-labeled primary antibody-embedded zone 5.
  • Step 105 When the external sample diluent is detected by the photoelectric sensor (b2) provided on the liquid outlet of the enzyme-labeled anti-embedding area 5, the electromagnetic valve V1 is closed, the electromagnetic valve V5 is opened, and the positive pressure is sequentially generated by the plunger pump 19. The pressure and the negative pressure suction cause the serum, the external diluent, and the pre-embedded enzyme-labeled primary antibody to re-dissolve back and forth between the enzyme-labeled primary antibody-resistant region 5 and the first mixed conduit 6 to obtain a first mixed solution.
  • Step 106 The first mixed liquid is sucked into the magnetic target secondary anti-embedding area 7, and the first mixed liquid is combined with the antigen-antibody through the second mixing pipe 8, and the formed reactant is captured by the magnetic beads, and the magnetic beads are magnetically
  • the magnet outside the standard anti-embedding zone 7 is adsorbed and stabilized in the magnetic-labeled secondary anti-embedding zone 7, and the remaining reaction liquid is discharged from the chip through the liquid driving force inlet under the negative pressure suction of the plunger pump 19, and then proceeds to the next Cleaning step.
  • Step 107 Close the electromagnetic valve V5, and open the electromagnetic valve V3, so that the external cleaning liquid enters the magnetic standard secondary anti-embedding area 7, and passes the photoelectric induction provided on the liquid inlet and the liquid outlet of the magnetic standard secondary anti-embedding area 7.
  • the devices (c1, c2) control the amount of injection of the cleaning liquid.
  • Step 108 After the external cleaning liquid and the magnetic beads are repeatedly cleaned, the magnets 25a and 25b adsorb the magnetic beads, and the negative pressure suction force is generated by the plunger pump, and the cleaned liquid is sucked and discharged into the external waste liquid pool 24.
  • Step 109 Close the electromagnetic valve V3, open the electromagnetic valve V2, and let the external substrate illuminating liquid enter the magnetic standard secondary anti-embedding area 7, and control the injection amount of the substrate luminescent liquid through the photoelectric sensors (c1, c2).
  • Step 110 After the substrate luminescent solution and the antigen antibody on the magnetic beads are fully reacted, the reaction solution is obtained, and the reaction solution is transported to the chemiluminescence detection zone 9 to complete the chemiluminescence detection; wherein the chemiluminescence detection zone 9 inlet port
  • the photoelectric sensors (d1, d2) provided on the liquid outlet are used to detect the capacity and position of the reaction liquid.
  • the reaction principle between the substances in the chemiluminescent microfluidic chip of the present embodiment is the same as that of the magnetic particle immunochemiluminescence reaction, that is, the antigen passing through the sample and the enzyme-labeled primary antibody (the primary antibody is labeled with a catalytic group such as HRP or AP). Binding, and then combined with the magnetic secondary antibody (the secondary antibody is immobilized on the magnetic beads) to form a double-anti-sandwich composite, the magnet adsorbs the magnetic beads, washes away the unbound antigen and the enzyme-labeled primary antibody, and adds the substrate reaction solution, one
  • the enzyme group such as HRP or AP which is resistant to the labeling catalyzes the luminescence of the substrate reaction solution.
  • the intensity of the luminescence is directly proportional to the amount of antigen.
  • the present invention provides a liquid drive apparatus that can implement the functions described in Embodiment 1 or 2.
  • the liquid driving device is a plunger pump 19.
  • the liquid driving device can be provided in various types, such as an existing syringe pump, a diaphragm pump, a peristaltic pump, etc., and any predetermined area capable of driving the liquid under pressure to the chip should fall into the present.
  • the scope of protection of the invention Although the syringe pump, diaphragm pump, and peristaltic pump can drive the liquid flow, they do not control the liquid to stay at a specific position, and the plunger pump can better solve this problem.
  • a plunger pump suitable for use in the present invention may be a plunger pump well known to those skilled in the art, which typically includes a pump chamber 194 and a plunger 193 having a liquid inlet 191 and a liquid outlet 192, a plunger The top end of the 193 is inserted into the pump chamber, and the plunger 193 reciprocates in the axial direction along the inner wall of the pump chamber 194; the inlet port 191 and the liquid outlet 192 are respectively provided with valves V4 and V6. Since the plunger pump is used more for liquid absorption and drainage, the two ports provided on the pump chamber are often referred to as "inlet and outlet", but it should be noted that the "inlet” here The mouth and the liquid outlet are not limited to the liquid inlet and the outlet.
  • the valve V4 at the inlet port 191 is opened, and the plunger moves downward, and the liquid is approaching.
  • the pressure at one end of the plunger pump inlet port 191 becomes small, causing a pressure difference between the two ends of the liquid, and the liquid moves toward the inlet port 191 under the pressure difference.
  • the outlet port is opened.
  • the valve V6 makes the inside of the chip communicate with the outside atmosphere, and the air on both sides of the liquid is on both sides of the air (the air on one side enters the inside of the chip through the liquid outlet and the liquid inlet, and the air on the other side can flow from the air into the port (such as Under the action of the inlet or the separately set air to the inside of the chip, the pressure is balanced and the liquid can stay at the predetermined position.
  • the present invention provides a liquid metering chamber that can perform the functions described in Embodiments 1 or 2.
  • the liquid quantitative chamber of the present invention can realize that “the liquid to be quantitatively flows from the liquid inlet of the liquid quantitative chamber into the liquid quantitative chamber, and after filling the liquid quantitative chamber to reach the liquid outlet”, the shape thereof
  • the structure can be selected as needed, and the present invention is not limited thereto, and for example, it may be a pipe shape, a polygonal shape, or the like.
  • the liquid metering chamber is a chamber of a hexagonal structure.
  • the liquid inlet and the liquid outlet of the liquid metering chamber are respectively two opposite corners of the hexagonal structure; the angles of the two diagonals are less than 120°.
  • the liquid inlet of the liquid metering chamber has a width of 0.3-3 mm (preferably 0.8-1.5 mm) and a height of 0.3-3 mm; and the outlet of the liquid metering chamber has a width of 0.3-3 mm (preferably 0.8-1.5mm), height 0.3-3mm.
  • the width of the inlet is too wide or too narrow, and the height is too high or too low, it is not conducive to the quantitative operation.
  • the width of the inlet is too wide or the height is too high, it is easy to cause the liquid to not fill the liquid quantitative chamber and flow to it.
  • Liquid port which can not achieve accurate liquid quantification, and when the inlet port width is too narrow or the height is too low, the corresponding length needs to be increased to meet the volume requirement, which may lead to an increase in chip length and an increase in chip volume. .
  • the surface of the liquid metering chamber is a surface formed by a hydrophilic surface modification;
  • the liquid inlet chamber has a width of 0.3-5 mm and a height of 0.3-3 mm; and the liquid quantitative chamber is out.
  • the liquid port has a width of 0.3 to 5 mm and a height of 0.3 to 3 mm.
  • Hydrophilic surface modifications include, but are not limited to, plasma, hydroxylation, carboxylation modifications.
  • the hydrophilic modification of the surface of the liquid quantification chamber is more conducive to the filling of the liquid in the cavity.
  • the width of the liquid inlet port and the liquid outlet of the liquid dosing chamber can be appropriately increased, thereby reducing the liquid quantification.
  • the length of the chamber and chip is a surface formed by a hydrophilic surface modification;
  • the liquid inlet chamber has a width of 0.3-5 mm and a height of 0.3-3 mm; and the liquid quantitative chamber is out.
  • the liquid port has a width of 0.3 to 5 mm and
  • the surface of the liquid quantitative chamber is a surface formed by a hydrophobic surface modification
  • the liquid inlet chamber has a width of 0.3-2 mm and a height of 0.3-3 mm; the liquid quantitative chamber is discharged.
  • the mouth has a width of 0.3-2 mm and a height of 0.3-3 mm.
  • Hydrophobic modifications include, but are not limited to, hydrophobic physical modifications, hydrophobic chemical modifications (e.g., nanoparticle coatings, extended chain alkyl groups, etc.).
  • the surface of the liquid metering chamber is modified by a hydrophobic surface to prevent the liquid from hanging on the wall, and to ensure that the liquid reaches the liquid outlet after being filled in the liquid metering chamber.
  • the present invention provides a liquid identification site and liquid identification device that can perform the functions described in Embodiment 1 or 2.
  • the liquid identification site is used for positioning or fixing the liquid identification device.
  • the present invention does not limit the structure of the liquid identification device as long as the liquid can be recognized.
  • the liquid sensing device disclosed in the patent application of the publication No. 105214744A can be used as the liquid identification device of the present invention, but such a liquid sensing device has a complicated structure, and the conductive needle needs to be built into the inside of the chip, and the conductive pin and the conductive needle The contact of the reaction liquid may affect the experimental results under certain circumstances, and the chip preparation is difficult. In this embodiment, a more preferable liquid identification device is provided.
  • the liquid identification site is used to position the liquid identification device, and the liquid identification device includes a light source generation module 28 and a photoelectric sensor 29; the liquid recognition site includes an upper point for positioning the light source generation module 28 and is used for positioning The lower point, the upper point and the lower point of the photoelectric sensor 29 are respectively arranged on the outer side of the chip body, and the upper point, the corresponding liquid inlet or the liquid outlet and the lower point are arranged in a vertical line.
  • the light source generating module 28, the corresponding liquid inlet or the liquid outlet, and the first photoelectric sensor 29 are arranged in a vertical line.
  • the "corresponding liquid inlet or outlet” herein corresponds to the liquid metering chamber or the detection chamber.
  • a liquid inlet or a liquid outlet for example, when a liquid identification device is provided at a liquid outlet of the liquid metering chamber, the light source generating module, the liquid outlet of the liquid metering chamber, and the photoelectric sensor are arranged in a vertical line;
  • the light source generating module, the liquid inlet port of the liquid metering chamber, and the photoelectric sensor are arranged in a vertical line;
  • the liquid outlet of the sample metering chamber is provided with the liquid identifying device, The light source generating module, the liquid outlet of the sample quantitative chamber, and the photoelectric sensor are arranged in a vertical line.
  • Optical sensing is used to identify, quantify and control liquids. Compared with conductive contact methods, this method reduces the intervention of metal on the intra-chip reaction system, improves the detection efficiency, and thus improves the accuracy of quantification.
  • the identification device can be disposed outside the microfluidic chip, and is convenient to be fixed in the instrument without being disposed on the chip, thereby reducing the processing difficulty of the chip. When using, simply align the light source generation module and the photoelectric sensor with the liquid recognition site.
  • the chip body includes a top plate 1 and a bottom plate 20; the top plate 1 and the bottom plate 20 are laminatedly connected; a connection of the top plate 1 and the bottom plate 20 is provided with a main fluid channel and a plurality of functional chambers; and the light source generating module 28 is disposed to be positioned and liquid quantitatively Immediately above the corresponding position of the top plate 1 corresponding to the liquid inlet or the liquid outlet of the chamber, the photosensor 29 is positioned directly below the corresponding position of the bottom plate 20 corresponding to the liquid inlet or the liquid outlet of the liquid metering chamber. .
  • the light source generating module is a module capable of providing a light source, which may be an LED, a halogen lamp, a laser lamp or the like. Under the illumination of the light source, due to the difference in transmittance and refractive index of the gas and liquid to the light, the intensity of the light irradiated to the photoelectric sensor is different, and the photoelectric sensor can recognize the gas and the liquid, thereby discriminating whether the liquid reaches the sensing point. . When the liquid flows to the liquid inlet or the liquid outlet, the liquid identification device can perform rapid identification to control the liquid driving device.
  • An embodiment of the present invention also provides an analytical instrument having a microfluidic chip, comprising an instrument frame, at least one reagent storage pool, a liquid driving device, a detecting device, and a microfluidic chip in any of the above embodiments; a microfluidic chip is mounted in the instrument frame; the liquid driving device is connected to the liquid driving force inlet of the microfluidic chip; the reagent storage pool and the corresponding reagent inlet are communicably connected; the detecting device is configured to receive the processing microfluid The detection signal sent by the control chip.
  • the liquid driving device is a plunger pump; each of the reagent storage pools is provided with an opening communicating with outside air.

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Abstract

一种微流控芯片，包括芯片主体、以及设置在该芯片主体上的进样口（2）、液体驱动力入口（13）、主流体通道和多个功能腔室；多个功能腔室中的至少一个为液体定量腔室（4，5，7），其具有预定的容积，并且在液体定量腔室（4，5，7）的出液口处设置有液体识别位点，需定量的液体从该液体定量腔室（4，5，7）的进液口流入该液体定量腔室（4，5，7），充满该液体定量腔室（4，5，7）后到达出液口。该微流控芯片通过特定的液体定量腔室来实现液体的识别定位和定量，降低了芯片的制作工艺难度，提高了定量的准确性。

Description

一种微流控芯片及具有该微流控芯片的分析仪器 技术领域
本发明涉及医疗器械领域，尤其涉及一种微流控芯片及具有该微流控芯片的分析仪器。
背景技术
体外诊断(In Vitro Diagnosis,IVD)是指从人体中取出样本(血液、体液、组织等)进行检测分析从而对疾病进行诊断，检测过程中需要相应的仪器和试剂，而这些仪器和试剂就组成了体外诊断系统。体外诊断的系统大致分为两种；一种是以检测中心实验室为代表，它具有系统模块化、自动化，流水线式的进行样本检验，从而也具有高通量、高效率、高敏感度的优势，但是整套系统也有费用昂贵，所占体积大，需要专业人员操作的缺陷，它主要应用于大型医院。另外一种是以即时检测(point-of-care testing，POCT)为代表，它的系统具有集成化、小型化、随时随地进行样本检验，从而也具有价格实惠、操作简单，结果报告及时的优势，但是它的测试结果和中心实验室相比还存在灵敏度，稳定性不高的缺点。
对于POCT，国内外都有把微流控技术运用到体外诊断的产品中。微流控(microfluidics)是在一块具有微管道的芯片上对微流体进行控制操作的一门交叉学科，涉及到生物、化学、流体物理、电子、光学、机械工程等领域。微流控装置通常被称为微流控芯片，也被称为芯片实验室(Lab on a Chip)。通常把生物、化学、医学分析过程的样品制备、反应、分离、检测等基本操作集中在一块芯片上，完成一个系统功能。现有的微流控芯片主要以定性检测为主，定量检测的微流控芯片较少，且现有的定量微流控芯片制备复杂，生产效率低，如公开号为“CN105214744”的中国专利申请公开了“一种磁微粒化学发光微流控芯片”，所述微流控芯片包括顶板和底板，其中，所述顶板包含气泵、加样口、样本填充区、标记配体存储池和样本混合区；所述底板包含过滤区、磁微粒包被区、清洗区、检测区、清洗液存储池、发光基底液存储池和液体释放通道；所述顶板和底板都包含液体传感装置，用来确定微流控芯片内液体的流动状态以及是否混入气泡等，此专利中的芯片采用多层结构，且其采用特定容积的容纳袋来实现液体的定量，此种定量结构虽然简单，但容纳袋表面极易出现液体挂袋现象(即将液体从容纳袋中压出时，部分液体挂留在袋中，无法保证将液体全部压出)，且容纳袋每次受到挤压的变形量都不一样，所以每次残留在容纳袋中的液体量不一致，进而液体被挤出的液体量不一 样，特别是需要小量液体的时候，容纳袋的误差更加大，相对于微流控芯片，需要的都是几十微升的量，所以容纳袋的定量精准度无法达到要求，定量准确性较差，从而影响检测结果，同时容纳袋需要内置到芯片中，增加了芯片的生产难度。
发明内容
为解决上述问题，一方面本发明提供了一种微流控芯片，其可以实现定量检测，且结构简单，降低了芯片的制作工艺难度。
本发明采用的技术方案为：一种微流控芯片，包括芯片主体、以及设置在所述芯片主体上的进样口、液体驱动力入口、主流体通道和多个功能腔室；
所述主流体通道连通所述多个功能腔室，所述进样口和所述液体驱动力入口分别与所述主流体通道连通，所述液体驱动力入口用于连接液体驱动装置以驱动液体流入或流出功能腔室；
所述多个功能腔室中的至少一个为液体定量腔室；所述液体定量腔室具有预定的容积，且在液体定量腔室的出液口处设置有液体识别位点，需定量的液体从所述液体定量腔室的进液口流入所述液体定量腔室，充满所述液体定量腔室后到达所述出液口。
在其中一个实施例中，所述液体定量腔室包括试剂定量腔室，所述试剂定量腔室的进液口与试剂支通道的一端连通，所述试剂支通道的另一端与试剂入口连通。
在其中一个实施例中，所述液体定量腔室的进液口处也设置有液体识别位点。
在其中一个实施例中，所述液体驱动装置为柱塞泵。
在其中一个实施例中，所述液体定量腔室还包括样品定量腔室，液体样品经所述进样口流入所述样品定量腔室进行定量；所述样品定量腔室位于所述试剂定量腔室的上游；
所述微流控芯片上还设有空气入口及与其连通的空气支通道，所述空气支通道的一端与所述空气入口连通，另一端与所述样品定量腔室和所述进样口之间的主流体通道连通，所述空气支通道的另一端与所述主流体通道的连通处邻近所述所述样品定量腔室。
在其中一个实施例中，所述功能腔室包括检测腔室，所述检测腔室具有预定的容积，且在所述检测腔室的出液口处设置有液体识别位点，待检测的液体经所述检测腔室的进液口流入所述检测腔室，充满所述检测腔室后到达出液口。
在其中一个实施例中，所述检测腔室的进液口处也设置有液体识别位点。
在其中一个实施例中，所述液体识别位点用于定位液体识别装置；所述液体识别装置包括光源生成模块和光电感应器；
所述液体识别位点包括用于定位所述光源生成模块的上位点和用于定位所述光电感应器的下位点，所述上位点和所述下位点分别设于所述芯片主体的外侧，所述上位点和下位点的位置与相应的出液口或进液口对应，以使得定位后的所述光源生成模块、相应的出液口或进液口、所述光电感应器呈垂直线依次布设。
在其中一个实施例中，所述液体定量腔室为六边形结构的腔室。
在其中一个实施例中，所述液体定量腔室的进液口的宽度为0.3-3mm，高度为0.3-3mm；所述液体定量腔室的出液口的宽度为0.3-3mm，高度为0.3-3mm；或
所述液体定量腔室的表面为经亲水性表面修饰后而形成的表面；所述液体定量腔室的进液口的宽度为0.3-5mm，高度为0.3-3mm；所述液体定量腔室的出液口的宽度为0.3-5mm，高度为0.3-3mm；或
所述液体定量腔室的表面为经疏水性表面修饰后而形成的表面，所述液体定量腔室的进液口的宽度为0.3-2mm，高度为0.3-3mm；所述液体定量腔室的出液口的宽度为0.3-2mm，高度为0.3-3mm。
在其中一个实施例中，所述芯片主体包括顶板和底板；所述顶板与所述底板层叠连接，所述顶板与所述底板的连接处设置有所述主流体通道和所述多个功能腔室。
在其中一个实施例中，所述底板为光滑的平板，所述顶板上设置有微孔、微通道或微腔体以与所述底板形成所述进样口、液体驱动力入口、主流体通道或功能腔室。
在其中一个实施例中，所述进样口和所述液体驱动力入口分别设置在所述主流体通道的两端。
另一方面，本发明还提供了一种具有微流控芯片的分析仪器，其包括仪器框架、至少一个试剂存储池、液体驱动装置、检测装置和以上所述的微流控芯片；其中，所述微流控芯片安装在所述仪器框架中；所述液体驱动装置与微流控芯片的液体驱动力入口相连；所述试剂存储池与对应的试剂入口可通断地连通；所述检测装置用于接收处理微流控芯片发出的检测信号。
在其中一个实施例中，所述液体驱动装置为柱塞泵；每个所述试剂存储池上均设有与外界空气连通的开口。
相对于现有技术，本发明具有如下有益效果：
本发明提供的微流控芯片通过特定的液体定量腔室结合液体驱动装置来实现液体的定量，相对于现有通过挤压镶嵌于芯片中试剂包来实现定量的技术，本发明的液体定量腔室，提高了定量的准确性；而且试剂可外置于芯片，相对于现有技术中多层芯片结合和试剂包镶嵌于芯片中，降低了芯片的制作工艺难度，提高了检测的准确性。
本发明的微流控芯片的芯片主体可包括层叠设置的顶板和底板，需要加工完成的结构均可设置的顶板上，底板仅为光滑的平板，这样可进一步降低芯片的制作工艺难度，提高生产效率。
附图说明
图1是本发明提供的微流控芯片的一种实施例的结构示意图；
图2是本发明提供的液体识别装置的截面示意图；
图3是本发明提供的微流控芯片的一种实施例的传感器设置结构图；
图4是本发明提供的微流控芯片使用时磁铁设置位置的截面示意图；
图5是本发明提供的液体驱动装置的一种实施例的结构示意图；
其其中，1、顶板；2、进样口、3、全血过滤区；4、样品定量区；5、酶标一抗包埋区；6、第一混匀通道；7、磁标二抗包埋区；8、第二混匀通道；9、化学发光检测区；10、稀释液入口；11、底物发光液入口；12、清洗液入口；13、液体驱动力入口；14、空气入口；15、密封垫；16、稀释液支通道；17、底物发光液支通道；18、清洗液支通道；19、柱塞泵；20、底板；21、稀释液存储池；22、底物发光液存储池；23、清洗液存储池；24、废液池；25a/25b、磁铁；26、磁珠；27、空气支通道；28、光源生成模块；29、光电感应器；191、柱塞泵的进液口；192、柱塞泵的出液口；193、柱塞；194、泵室。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
实施例1
本实施例提供了一种微流控芯片，该微流控芯片包括芯片主体、以及设置在所述芯片主体 上的进样口、液体驱动力入口、主流体通道和多个功能腔室；下面进行详细说明。
在本实施例中，主流体通道连通多个功能腔室，以引导流体在功能腔室之间的流动。
在本实施例中，功能腔室至少具有容纳的功能，优选地，功能腔室除具容纳功能外，还具有其他功能，其他功能可以是功能腔室内实现的，也可以是功能腔室结合其外部的必要部件(这些必要的部件可在芯片外部固定，无需设置在芯片内或其表面)共同实现的。
在本实施例中，进样口和液体驱动力入口分别与主流体通道连通，液体驱动力入口用于连接液体驱动装置以驱动液体流入或流出功能腔室，进样口用于将液体样品引入主流体通道中，液体样品经主流体通道进入各功能腔室。
在本实施例中，多个功能腔室中的至少一个为液体定量腔室；所述液体定量腔室具有预定的容积，且在液体定量腔室的出液口处设置有液体识别位点，需定量的液体从所述液体定量腔室的进液口流入所述液体定量腔室，充满所述液体定量腔室后到达所述出液口。液体识别位点用于定位或固定液体识别装置，液体识别装置用于识别液体，当液体流至液体识别位点处时，液体识别装置可识别液体并提供液体到达信号；因此当液体到达出液口处时，液体识别装置可提供液体到达信号，指示液体已将液体定量腔室充满，此时控制液体驱动装置停止驱动液体，实现液体在液体定量腔室中的定量。
本实施例提供的微流控芯片通过特定的液体定量腔室结合液体驱动装置来实现液体的定量，相对于现有通过挤压镶嵌于芯片中试剂包来实现定量的技术，本发明的液体定量腔室，提高了定量的准确性。试剂可外置于芯片，相对于现有技术中多层芯片结合和试剂包镶嵌于芯片中，降低了芯片的制作工艺难度，提高了检测的准确性。
应当说明的是，主流体通道和多个功能腔室可以通过激光加工、模型注塑加工等多种方式在芯片主体内部成形，也可通过设置为分离式的顶板和底板，在顶板或底板上加工出特定形状，然后相互封装在一起；由于前一种加工方式较为繁琐，在一个优选的实施例中，所述芯片主体包括顶板和底板；顶板与底板层叠连接；顶板与底板的连接处设置有所述主流体通道和多个功能腔室；更优选地，底板为光滑的平板，顶板上设置微孔、微通道或微腔体以与底板配合形成进样口、液体驱动力入口、主流体通道或多个功能腔室，这样的微流控芯片制备起来更为方便，降低了生产工艺难度，只需在顶板上加工所需的特定结构即可，提高了生产效率。具体地，所述底板为光滑的平板，所述顶板上设有多个微通道以与所述底板结合形成主流体通道，所述顶板上设有多个微腔以与所述底板结合形成所述多个功能腔室，所述顶板上设有多个孔以与所述 底板结合形成所述进样口或液体驱动力入口；为了便于进样，进样口的尺寸通常大于其他入口的尺寸。
液体定量腔室的数量、其定量的液体类型(例如液体样品、反应试剂、样品处理试剂等)、设置位置以及其他功能腔室的类型可根据实际需要进行选择。
优选地，液体定量腔室包括试剂定量腔室，试剂定量腔室的进液口与试剂支通道的一端连通，试剂支通道的另一端与试剂入口连通，试剂经试剂入口、试剂支通道进入试剂定量腔室进行定量。在利用微流控芯片进行定量检测时，液体样品(即从进样口流入的待测液体)和试剂(例如反应试剂、样品处理试剂等)的量需要进行定量，通常试剂的定量需要在芯片内部完成，液体样品可选择地在微流控芯片外实现定量，本实施例中试剂定量腔室即用于试剂的定量。此处的“液体定量腔室包括试剂定量腔室”应理解为液体定量腔室应至少具有用于定量试剂的试剂定量腔室这种类型，当然也可以进一步包括用于定量其他液体类型的液体定量腔室，如用于定量液体样品的样品定量腔室等。
试剂定量腔室的数量可为一个、两个或多个，可根据微流控芯片的实际需要进行选择。例如当微流控芯片为化学发光微流控芯片时，为了实现化学发光定量检测，试剂定量腔室应至少设置一个以用于定量底物发光液，其余的反应必须的物质如酶标一抗、磁标二抗可分别包埋在酶标一抗包埋腔室和磁标二抗包埋腔室这两个功能腔室内；优选地，磁标二抗包埋腔室为试剂定量腔室，其内不仅包埋有磁标二抗，还可用于定量底物发光液；更优选地，磁标二抗包埋腔室还用于磁珠清洗。
可选地，试剂入口与试剂储存池通过阀门可通断地连接，试剂存储池设有与外界空气连通的开口，试剂存储池设有开口可便于液体驱动装置将其内的液体引流至芯片内；为了便于芯片的制备，优选地，试剂存储池设于微流控芯片的外部，使用时将试剂存储池安装在试剂入口处，以将试剂引入芯片内。
可选地，液体定量腔室的进液口处也设置有液体识别位点。此液体识别位点同样可用于定位或固定液体识别装置，其设置可便于对芯片内液体的流动进行监控、控制，其还可实现两种定量液体之间的混合，比如液体样品与试剂之间的混合。在芯片内部，若要实现两种液体的混合，需要两种液体接触，中间不存在空隙，而本发明的微流控芯片若要同时实现液体的定量以及两种液体的接触，这就要求其中一种定量后的液体停留在预定位置处，另一种液体最好从此预定位置开始流入液体定量腔室，在液体定量腔室内实现定量，而此预定位置的最佳选择即液 体定量腔室的进液口处；在进液口处设置液体识别位点以定位液体识别装置，此液体识别装置即可提供其中一种液体的停留指示信号及另一种液体的进液信号，在液体定量腔室的出液口处的液体识别装置的配合下，即可实现液体的定量以及两种液体的接触。接下来，以液体样品与试剂的混合为例来具体说明下微流控芯片的一个使用方法：
微流控芯片使用时，试剂入口与试剂储存池通过阀门可通断地连接，试剂存储池设有与外界空气连通的开口。液体样品(为了便于与定量的试剂精确混合或反应，液体样品优选为预定量的液体样品)在液体驱动装置的作用下从进样口处经主流体通道流至试剂定量腔室的进液口处时，定位于试剂定量腔室的液体识别位点上的液体识别装置获取信号，控制液体驱动装置停止驱动作用，此时液体样品停止流动，液体样品的前端停留在进液口处；接着，关闭空气入口，打开试剂入口与试剂储存池之间的阀门，试剂在液体驱动装置的作用下从试剂入口经试剂支通道、试剂定量腔室的进液口进入试剂定量腔室内，当流至试剂定量腔室的出液口时，试剂充满试剂定量腔室，此时关闭试剂入口与试剂储存池之间的阀门，打开空气入口，试剂定量腔室定量的试剂及液体样品在液体驱动装置的作用下继续流动，并可在液体驱动装置的正压、负压交替作用下实现正向逆向运动而实现混合和/或反应。
为了减小试剂支通道对于试剂定量腔室进液口处的液体识别装置的影响，同时便于加工制作，具体地，所述试剂支通道的一端通过所述主流体通道与试剂定量腔室的进液口连通，所述试剂支通道与所述主流体通道的连通处邻近试剂定量腔室的进液口处，以使定量在可控范围内，减小定量误差，例如，所述试剂支通道与所述主流体通道的连通处距离试剂定量腔室的进液口的距离为0.5～10mm(优选为0.5-2mm)。
可选地，所述进样口和所述液体驱动力入口分别设置在所述主流体通道的两端。
可选地，所述功能腔室包括检测腔室，检测腔室具有预定的容积，且在检测腔室的出液口处设置有液体识别位点，待检测的液体流入检测腔室，充满检测腔室后到达出液口。检测腔室的出液口处设置的液体识别位点可用于定位或固定液体识别装置，当待检测的液体到达检测腔室的出液口处时，液体识别装置发出信号，液体驱动装置控制待检测的液体停止流动，此时即可进行检测。进一步地，检测腔室的进液口处也设有液体识别位点。
可选地，液体定量腔室还进一步包括样品定量腔室，液体样品经进样口流入样品定量腔室进行定量；样品定量腔室位于试剂定量腔室的上游；微流控芯片上还设有空气入口及与其连通的空气支通道，所述空气支通道的一端与所述空气入口连通，另一端与所述样品定量腔室和所 述进样口之间的主流体通道连通，所述空气支通道的另一端与所述主流体通道的连通处邻近所述样品定量腔室；此处“邻近”通常可理解为“距离样品定量腔室的进液口0.5～10mm(优选为0.5-2mm)”。
微流控芯片使用时，将空气入口与芯片外部的空气管道通过阀门可通断地连接，以控制空气进入芯片内部。液体样品在液体驱动装置的作用下经进样口从样品定量腔室的进液口流入样品定量腔室内，当液体样品流至样品定量腔室的出液口处时，即充满样品定量腔室，此时位于出液口的液体识别位点上定位的液体识别装置发出指示信号，控制空气入口打开，由于空气支通道中空气的流动所需驱动力小，而液体样品的流动所需驱动力更大，因此液体样品停留在空气支通道与主流体通道的连通处不再继续流入样品定量腔室，即可完成液体样品在样品定量腔室中的定量。定量后的液体样品可在液体驱动装置的作用下继续流至试剂定量腔室的进液口处，待试剂定量腔室完成试剂的定量后(过程如上所述)，液体样品和试剂在液体驱动装置的正压、负压交替作用下实现混合和/或反应。
本实施例中的微流控芯片，由于设置了样品定量腔室，可便于液体样品的定量，而无需在芯片外另行定量，使得芯片使用更为方便。
可选地，所述液体样品为全血，所述进样口与所述样品定量腔室之间设有全血过滤腔室，所述全血过滤腔室中设有全血滤膜；当微流控芯片用于临床诊断时，全血是常见的检测样品，检测时通常需要进行全血分离以将全血中的血清或血浆分离出来，再与试剂进行反应；芯片中设置全血过滤腔室，便于检测使用，同时相较于先定量全血，再进行全血分离的方式，在进样口与所述样品定量腔室之间设有全血过滤腔室，可通过样品定量腔室直接定量血清或血浆的用量，测量结果更为精确。所述全血滤膜的材质可为玻璃纤维、棉短绒纤维、聚酯纤维、纤维或混纺纤维；可选地，所述全血过滤滤垫的厚度为0.2-2.5mm；所述全血过滤滤垫的吸附速度为4-150s/4cm，吸水性为30-250mg/cm 2。
可选地，全血过滤区的出液口为三角形出液口；全血过滤区面积为30-300mm 2，宽为2-20mm，长为5-25mm，深为0.3-3mm，前端三角形的角度为15-160℃。
实施例2
请参照图1～图5，本实施例提供了一种化学发光微流控芯片，其包括芯片主体、以及设置在芯片主体上的进样口2、液体驱动力入口13、底物发光液入口11、清洗液入口12、底物发光液支通道17、清洗液支通道18、主流体通道和多个功能腔室；下面进行详细说明。
在本实施例中，主流体通道连通多个功能腔室，以引导流体在功能腔室之间的流动。
功能腔室包括通过主流体通道依次连通的酶标一抗包埋区5、磁标二抗包埋区7和化学发光检测区9。
其中，酶标一抗包埋区5包埋有酶标一抗；磁标二抗包埋区7包埋有磁标二抗；磁标二抗包埋区7为液体定量腔室；液体定量腔室用于定量液体，待定量的液体(例如底物发光液)进入液体定量腔室后，可在液体定量腔室内实现定量(即得到所需用量的液体)，以与定量的液体样品或其他反应试剂反应，从而实现定量检测。
在本实施例中，液体定量腔室具有预定的容积，且在液体定量腔室的出液口处设置有液体识别位点，需定量的液体从液体定量腔室的进液口流入液体定量腔室，充满液体定量腔室后到达出液口；液体识别位点用于定位或固定液体识别装置，液体识别装置用于识别液体。当液体到达出液口处时，液体识别装置可提供液体到达信号，指示液体已将液体定量腔室充满，此时控制液体驱动装置停止驱动液体，即可实现液体在液体定量腔室中的定量。化学发光微流控芯片通过特定的液体定量腔室结合液体驱动装置来实现液体的定量，可提高了定量的准确性。
在本实施例中，化学发光检测区9用于容置化学发光反应产物，以与外部检测装置结合完成检测过程。
进样口2与液体驱动力入口13分别与主流体通道连通，驱动力入口13用于连接液体驱动装置以驱动液体流入或流出功能腔室；进样口2用于将液体样品引入主流体通道中，液体样品经主流体通道进入各功能腔室。
在本实施例中，底物发光液支通道17的一端与底物发光液入口11连通，其另一端与磁标二抗包埋区7的进液口连通，底物发光液经底物发光液入口11、底物发光液支通道17进入磁标二抗包埋区7进行定量。
清洗液支通道18的一端与清洗液入口12连通，其另一端与磁标二抗包埋区7的进液口连通，清洗液经清洗液入口12、清洗液支通道18进入磁标二抗包埋区7进行磁珠清洗。
本实施例的微流控芯片使用时，底物发光液入口11、清洗液入口12分别与底物发光液储存池22、清洗液储存池23通过阀门V2、V3可通断地连接，底物发光液储存池22、清洗液储存池23上分别设有与外界空气连通的开口；液体驱动装置安装在液体驱动力入口13处，用以驱动芯片内液体流动；磁标二抗包埋区7的外侧固定有磁铁(例如磁铁25a，25b)，以便固定磁珠26。磁标二抗包埋区为液体定量腔室，其可用于定量底物发光液，可选地，还可进一步 用于定量清洗液。
本实施例的微流控芯片的一工作方式如下：预定量的液体样品(如经稀释液稀释后的血清或血浆)在液体驱动装置的作用下从进样口2处经主流体通道流至酶标一抗包埋区5，与其中包埋的酶标一抗混合反应，其后反应液到达磁标二抗包埋区7，与其中包埋的磁标二抗继续混合反应，在磁珠上形成双抗夹心结构的反应物，磁珠被磁铁吸附，反应物在磁珠的作用下稳定在磁标二抗包埋区7内，而其余的反应液在液体驱动装置的作用下经液体驱动力入口13排出芯片；然后，关闭芯片上的空气流入端口(如样品进口)，打开清洗液储存池23与清洗液入口12之间的阀门V3，清洗液在液体驱动装置的作用下经清洗液支通道18进入磁标二抗包埋区7以对其中的磁珠进行清洗，当磁标二抗包埋区7完成对清洗液的定量时，即可关闭清洗液储存池23与清洗液入口12之间的阀门V3，打开空气流入端口，清洗后的液体在液体驱动装置的作用下经液体驱动力入口13排出芯片，为了保证清洗效果，可反复清洗数次(磁珠清洗方式不限于此处描述的方式，也可以通过例如在清洗液中移动磁铁的方式实现磁珠的清洗)；接着关闭芯片上的空气流入端口(如样品进口)，打开底物发光液储存池22与底物发光液入口11之间的阀门V2，底物发光液在液体驱动装置的作用下经底物发光液支通道19进入磁标二抗包埋区7，当磁标二抗包埋区7完成对底物发光液的定量时，关闭底物发光液储存池22与底物发光液入口11之间的阀门V2，液体驱动装置停止驱动作用，底物发光液不再流入磁标二抗包埋区7，打开芯片上的空气流入端口(如样品进口)，磁标二抗定量后的底物发光液与磁珠捕获的反应物进行发光反应，之后移除磁铁，磁标二抗包埋区7中的反应液在液体驱动装置的作用下流入化学发光检测区9进行检测。
上述化学发光微流控芯片结构紧凑，例如磁标二抗包埋区不仅用于包埋磁标二抗，其作为液体定量腔室还可用于定量底物发光液，而无须再另行设置液体定量腔室，磁标二抗包埋区还可进一步作为用于磁珠清洗的区，而无须再另行设置磁珠清洗区，大大节省了芯片的体积；同时，试剂存储池(如底物发光液存储池、清洗液存储池等)可外置于芯片，相对于现有技术试剂包镶嵌于芯片中，降低了芯片的制作工艺难度，提高了检测的准确性。
应当说明的是，主流体通道和多个功能腔室可以通过激光加工、模型注塑加工等多种方式在芯片主体内部成形，也可通过设置为分离式的顶板和底板，在顶板或底板上加工出特定形状，然后相互封装在一起；由于前一种加工方式较为繁琐，在一个优选的实施例中，芯片主体包括顶板1和底板20；顶板1与底板20层叠连接；顶板1与底板20的连接处设置有主流体通道和多个功能腔室；更优选地，底板20为光滑的平板，顶板20设置微孔、微通道或微腔体以与 底板配合形成进样口2、液体驱动力入口13、底物发光液入口11、清洗液入口12、底物发光液支通道17、清洗液支通道18、主流体通道或多个功能腔室，这样的微流控芯片制备起来更为方便，进一步降低了生产工艺难度，只需在顶板上加工所需的特定结构即可，进一步提高了生产效率。在一个实施例中，底板20为光滑的平板，顶板1上设有多个微通道以与底板20结合形成主流体通道，顶板1上设有多个微腔以与底板20结合形成多个功能腔室，顶板1上设有多个孔以与底板20结合形成进样口2、液体驱动力入口13、底物发光液入口11和清洗液入口12；为了便于进样，进样口2的尺寸通常大于其他入口的尺寸。
因此，上述化学发光微流控芯片的芯片主体可包括层叠设置的顶板和底板，需要加工完成的结构均可设置的顶板上，底板仅为光滑的平板，这样可进一步降低芯片的制作工艺难度，提高生产效率。
可选地，液体定量腔室的进液口处也设置有液体识别位点。此液体识别位点的设置可便于对芯片内液体的流动及可能存在的气泡进行监控、控制，其还可实现两种定量液体之间的混合，比如液体样品与试剂(如反应试剂、样品处理试剂等)之间的混合。进一步地，酶标一抗包埋区5也为液体定量腔室，芯片主体上还设有稀释液入口10和稀释液支通道16；稀释液支通道16的一端与稀释液入口10连通，另一端与酶标一抗包埋区5的进液口连通，样品稀释液经稀释液入口、稀释液支通道进入酶标一抗包埋区5进行定量。更进一步地，酶标一抗包埋区5的进液口和出液口处分别设置有液体识别位点，需定量的液体从其进液口流入酶标一抗包埋区5，充满酶标一抗包埋区5后到达出液口。样品稀释液不仅能够稀释液体样品(如血清、血浆等)，降低其浓度及粘稠度，其中含有的物质还可降低液体样品的本底值，使得检测更为精确，同时样品稀释液可以更好的复溶酶标一抗；在此技术方案中，酶标一抗包埋区可用于定量样品稀释液，而无需在芯片外部实现样品稀释液的定量，定量的样品稀释液可在酶标一抗包埋区与定量的液体样品进行混合，可节省人力，操作更加便捷。使用时，将稀释液入口10与稀释液储存池21通过阀门V1可通断地连接，稀释液储存池21上设有与外界空气连通的开口；预定量的液体样品(如经稀释液稀释后的血清或血浆)在液体驱动装置的作用下从进样口2处经主流体通道流至酶标一抗包埋区5的进液口处，关闭芯片上的空气入口(如样品进口)，打开稀释液储存池21与稀释液入口10之间的阀门V1，样品稀释液在液体驱动装置的作用下经稀释液支通道16进入酶标一抗包埋区5，当其充满酶标一抗包埋区5，到达酶标一抗包埋区5的出液口处时，关闭稀释液储存池21与稀释液入口10之间的阀门V1，打开空气流入端口(如样品进口)，液体样品和样品稀释液即可在液体驱动装置的负压作用下继续流动，并可在液体驱 动装置的正压、负压交替作用在主流体通道、酶标一抗包埋区5中实现混合，当然也可通过设置的混合通道实现更好的混合。
可选地，化学发光检测区9具有预定的容积，且在化学发光检测区9的进液口和出液口处分别设置有液体识别位点，待检测的液体经化学发光检测区9的进液口流入化学发光检测区9，充满化学发光检测区9后到达出液口，化学发光检测区9的容积小于等于磁标二抗包埋区7的容积。化学发光检测区9的出液口处设置的液体识别位点可用于定位或固定液体识别装置，当底物发光液与磁珠捕获的反应物进行反应后的反应液到达化学发光检测区的出液口处时，液体识别装置发出信号，液体驱动装置控制反应液停止流动，此时即可进行检测。
可选地，为了便于液体样品、试剂(样品稀释液、底物发光液等)之间的混合，主流体通道包括第一混匀通道6和第二混匀通道8；第一混匀通道6设于酶标一抗包埋区5和磁标二抗包埋区7之间；第二混匀通道8设于磁标二抗包埋区7和化学发光检测区9之间。
可选地，进样口2与液体驱动力入口13分别设置在主流体通道的两端。
如图4所示，可选地，为了便于固定磁珠，芯片主体与磁标二抗包埋区7对应的位置处设置有磁铁固定位点；进一步地，由于磁珠的清洗可在磁标二抗包埋区7进行，为了更好地实现磁珠清洗，磁标二抗包埋区7的上方和下方各布设一个用于定位磁铁25a，25b的磁铁固定位点，两个磁铁25a，25b对应于磁标二抗包埋区7的斜对角布设。
可选地，液体驱动装置为柱塞泵19，实施例3中关于柱塞泵的描述适用于本实施例。
可选地，功能腔室还包括样品定量腔室4，样品定量腔室4也为液体定量腔室，液体样品经进样口流入样品定量腔室4进行定量；样品定量腔室4位于酶标一抗包埋区5的上游；微流控芯片上还设有空气入口14及与其连通的空气支通道27，空气支通道27的一端与空气入口14连通，另一端与样品定量腔室4和进样口2之间的主流体通道连通，空气支通道27的另一端与主流体通道的连通处邻近样品定量腔室4。此处“邻近”通常可理解为“距离样品定量腔室4的进液口1～10mm”。通过设置了样品定量腔室，可便于液体样品的定量，而无需在芯片外另行定量，使得芯片使用更为方便。进一步地，样品定量腔室4的出液口处设置有液体识别位点，需定量的液体从其进液口流入样品定量腔室4，充满样品定量腔室4后到达出液口。更进一步地，样品定量腔室4的进液口处也设置有液体识别位点。
微流控芯片使用时，将空气入口与芯片外部的空气管道通过阀门可通断地连接，以控制空气进入芯片内部。液体样品在液体驱动装置的作用下经进样口从样品定量腔室的进液口流入样 品定量腔室内，当液体样品流至样品定量腔室的出液口处时，即充满样品定量腔室，此时位于出液口的液体识别位点上定位的液体识别装置发出指示信号，控制空气入口打开，由于空气支通道中空气的流动所需驱动压力小，而液体样品的流动所需驱动的压力更大，因此液体样品停留在空气支通道与主流体通道的连通处不再继续流入样品定量腔室，即可完成液体样品在样品定量腔室中的定量。定量后的液体样品可在液体驱动装置的作用下继续流至酶标一抗包埋区。
可选地，液体样品为全血，进样口7与样品定量腔室4之间设有全血过滤区3，全血过滤区3中设有全血滤膜；当微流控芯片用于临床诊断时，全血是常见的检测样品，检测时通常需要进行全血分离以将全血中的血清或血浆分离出来，再与试剂进行反应；芯片中设置全血过滤区，便于检测使用，同时相较于先定量全血，再进行全血分离的方式，在进样口与样品定量腔室之间设有全血过滤区，可通过样品定量腔室直接定量血清或血浆的用量，测量结果更为精确。全血滤膜的材质可为玻璃纤维、棉短绒纤维、聚酯纤维、纤维或混纺纤维；可选地，全血过滤滤垫的厚度为0.2-2.5mm；全血过滤滤垫的吸附速度为4-150s/4cm，吸水性为30-250mg/cm 2。
实施例4中关于液体定量腔室的描述适用于以上所述的液体定量腔室(包括磁标二抗包埋区7、酶标一抗包埋区5和样品定量腔室4)的描述，在此不再赘述。
实施例5中关于液体识别位点及液体识别装置的描述适用于以上所述的液体识别位点及液体识别装置的描述，在此不再赘述。
可选地，底物发光液支通道17的另一端与磁标二抗包埋区7的进液口的连通处位于磁标二抗包埋区7的进液口的主流体通道上；在一个实施例中，此处“邻近”理解为“距离磁标二抗包埋区7的进液口0.5～10mm(优选为0.5～2mm)”。
可选地，清洗液经清洗液入口12、清洗液支通道18进入磁标二抗包埋区7进行定量；清洗液支通道18的另一端与磁标二抗包埋区7的进液口的连通处位于与进液口邻近的主流体通道上；在一个实施例中，此处“邻近”理解为“距离磁标二抗包埋区7的进液口0.5～10mm(优选为0.5～2mm)”。优选地，清洗液支通道18的另一端与磁标二抗包埋区7的进液口的连通处在底物发光液支通道17的另一端与磁标二抗包埋区7的进液口的连通处的下游，这样可避免底物发光液被清洗液稀释。
可选地，稀释液支通道16的另一端与酶标一抗包埋区5的进液口的连通处位于与酶标一抗包埋区5的进液口邻近的主流体通道上；在一个实施例中，此处“邻近”理解为“距离酶标一抗包埋区5的进液口0.5～10mm(优选为0.5～2mm)”。
可选地，进样口2的体积为5ul-300ul。
可选地，全血过滤区3的出液口为三角形出液口；全血过滤区3面积为30-300mm 2，宽为2-20mm，长为5-25mm，深为0.3-3mm，前端三角形的角度为15-160℃。
可选地，样品定量腔室4的体积为1-50ul。
可选地，酶标一抗包埋区5的体积为5-50ul。
可选地，第一混匀管道6和第二混匀管道8的宽为200-2000um，长为5mm-40mm，深为0.2-3mm。
可选地，磁标二抗包埋区7的体积为10-200ul。
可选地，化学发光检测区9的体积为10-200ul。
接下来，结合图1～图5，描述根据本发明的一种实施方式的微流控芯片的检测方法。该方法包括步骤101至步骤110，各步骤具体如下：
步骤101：将分别与稀释液存储池21、底物发光液存储池22、清洗液存储池23、柱塞泵19、空气连通的钢针插入芯片中的密闭垫15，其中钢针分别与稀释液入口10、底物发光液入口11、清洗液入口12、液体驱动力入口13、空气入口14连接；将全血样本加入到进样口2，打开电磁阀V4并由柱塞泵19产生负压吸力，将全血样品吸入全血过滤区3。
步骤102：全血样品完成过滤后的血清被吸入到样品定量腔室4，并由样品定量腔室4进液口和出液口上设置的光电感应器(a1、a2)完成血清的定量测量。
当全血样品经过光电传感器a1上方是，感应器输出电压值发生变化，给系统一个识别信号，判断液体在芯片中的流动位置。当样品经过光电传感器a2时，判断样品把样品定量腔室4充满，该区域的固有体积即为样品的定量值。
步骤103：封堵进样口2并打开电磁阀V5，使得血清被吸入到酶标一抗包埋区5。
步骤104：当酶标一抗包埋区5的进液口上设置的光电感应器(b1)检测到血清时，关闭电磁阀V5，打开电磁阀V1，使得外部样品稀释液从电磁阀V1进入到酶标一抗包埋区5。
步骤105：当酶标一抗包埋区5出液口上设置的光电感应器(b2)检测到外部样品稀释液时，关闭电磁阀V1，打开电磁阀V5，并通过柱塞泵19依次产生正压和负压吸力，使得血清、外部稀释液、预先包埋的酶标一抗在酶标一抗包埋区5和第一混匀管道6之间来回流动复溶，获得第一混合液。
步骤106：第一混合液被吸入到磁标二抗包埋区7，并通过第二混匀管道8使第一混合液与抗原抗体结合，形成的反应物被磁珠捕捉，磁珠被磁标二抗包埋区7外侧的磁铁吸附而稳定 在磁标二抗包埋区7内，其余的反应液在柱塞泵19的负压吸力下经液体驱动力入口排出芯片，接着进行下一清洗步骤。
步骤107：关闭电磁阀V5，并打开电磁阀V3，使外部清洗液进入到磁标二抗包埋区7，并通过磁标二抗包埋区7进液口和出液口上设置的光电感应器(c1、c2)控制清洗液的注入量。
步骤108：在外部清洗液与磁珠反复清洗后，磁铁25a、25b吸附磁珠，通过柱塞泵产生负压吸力，将清洗过后的液体吸出排到外部废液池24中。
步骤109：关闭电磁阀V3，打开电磁阀V2，使外部底物发光液进入到磁标二抗包埋区7，并通过光电感应器(c1、c2)控制底物发光液的注入量。
步骤110：在底物发光液与磁珠上的抗原抗体充分反应后，获得反应液，反应液被运输到化学发光检测区9，以完成化学发光检测；其中，化学发光检测区9进液口和出液口上设置的光电感应器(d1、d2)用于检测反应液的容量及位置。
本实施例的化学发光微流控芯片中物质之间的反应原理同磁微粒免疫化学发光反应原理，即样品中的抗原通过和酶标一抗(一抗标记有HRP、AP等催化基团)结合，接着与磁标二抗(二抗被固定在磁珠上)结合形成双抗夹心复合物，磁铁吸附磁珠，清洗掉未结合的抗原和酶标一抗，加入底物反应液，一抗上标记的HRP、AP等酶基团催化底物反应液发光。发光强度和抗原的量成正比。
实施例3
请参考图5，本发明提供了可实现实施例1或2中所述功能的液体驱动装置。在本实施例中，液体驱动装置为柱塞泵19。
就结构而言，液体驱动装置可设置为多种，如现有的注射泵、隔膜泵、蠕动泵等，凡是能够实现将液体在压力作用下驱动至芯片内的预定区域，均应落入本发明的保护范围。注射泵、隔膜泵、蠕动泵虽然能够驱动液体流动，但它们不能很好地控制液体在特定位置停留，而柱塞泵能够较好地解决这个问题。适用于本发明的柱塞泵可为本领域技术人员所熟知的柱塞泵，其通常包括泵室194和柱塞193，泵室194上设有进液口191和出液口192，柱塞193的顶端插入泵室内，柱塞193沿着泵室194的内壁在其轴向上作往复运动；进液口191、出液口192处分别设有阀门V4、V6。由于柱塞泵被较多的应用于吸液、排液，泵室上设置的两个口通常被称为“进液口和出液口”，但需要说明的是，此处的“进液口和出液口”并不限于用于进液和出液，在本实施例中，柱塞泵工作时，进液口191处的阀门V4打开后，柱塞向下运动，此时液体临近柱塞泵进液口191的一端的压力变小，造成液体两端产生压力差，液体在压力差的作 用下向进液口191方向运动，当液体到达预定位置处时，打开出液口处的阀门V6，使得芯片内部与外部大气连通，液体两侧分别在两侧空气(其中一侧的空气经出液口、进液口进入芯片内部，另一侧的空气可从空气流入端口(如进样口或另行设置的空气至通道)进入芯片内部)的作用下，压力保持平衡，液体即可停留在预定位置处。
实施例4
请参考图1和图3，本发明提供了可实现实施例1或2中所述功能的液体定量腔室。
需要说明的是，本发明的液体定量腔室能够实现“需定量的液体从液体定量腔室的进液口流入液体定量腔室，充满液体定量腔室后到达出液口”即可，其形状结构可根据需要进行选择，本发明不对此作任何限制，例如其可为管道形状、多边形形状等。
实现“需定量的液体从所述液体定量腔室的进液口流入所述液体定量腔室，充满所述液体定量腔室后到达所述出液口”的方式有多种，例如控制液体定量腔室的宽度和高度、在液体定量腔室的表面进行亲疏水性处理等。
在本实施例中，所述液体定量腔室为六边形结构的腔室。可选地，所述液体定量腔室的进液口和出液口分别为所述六边形结构的两个对角；所述两个对角的角度小于120°。
可选地，液体定量腔室的进液口的宽度为0.3-3mm(优选为0.8-1.5mm)，高度为0.3-3mm；液体定量腔室的出液口的宽度为0.3-3mm(优选为0.8-1.5mm)，高度为0.3-3mm。进液口宽度过宽或过窄、高度过高或过低均不利于定量的进行，当进液口宽度过宽或高度过高时，容易造成液体无法充满液体定量腔室即流至其出液口，这样无法实现准确的液体定量，而当进液口宽度过窄或高度过低时，则需要相应增加长度来满足容积的要求，这样可能会导致芯片长度的增加及芯片体积的增大。
可选地，液体定量腔室的表面为经亲水性表面修饰后而形成的表面；液体定量腔室的进液口的宽度为0.3-5mm，高度为0.3-3mm；液体定量腔室的出液口的宽度为0.3-5mm，高度为0.3-3mm。亲水性表面修饰包括但不限于等离子、羟基化、羧基化修饰。液体定量腔室的表面进行亲水性修饰后，更有利于液体在腔体内的填充，此时可适当的加大液体定量腔室的进液口、出液口的宽度，从而可缩小液体定量腔室及芯片的长度。
可选地，液体定量腔室的表面为经疏水性表面修饰后而形成的表面，液体定量腔室的进液口的宽度为0.3-2mm，高度为0.3-3mm；液体定量腔室的出液口的宽度为0.3-2mm，高度为0.3-3mm。疏水性修饰包括但不限于疏水性物理修饰、疏水性化学修饰(如纳米粒子涂层、加 长链的烷基等)。液体定量腔室的表面经疏水性表面修饰后，可防止液体挂壁，而且可保证液体在液体定量腔室内充满后到达出液口。
实施例5
请参见图2，本发明提供了可实现实施例1或2中所述功能的液体识别位点及液体识别装置。
需要说明的是，液体识别位点用于定位或固定液体识别装置，本发明对于液体识别装置的结构不作限制，只要能实现液体的识别即可。如公开号为“105214744A”的专利申请中公开的液体传感装置即可作为本发明的液体识别装置，但这样的液体传感装置结构较为复杂，导电针需要内置入芯片内部，并且导电针与反应液体接触，在一定情况下会影响实验结果，且芯片制备难度较大，在本实施例中提供了一种更优选的液体识别装置。
在本实施例中，液体识别位点用于定位液体识别装置，液体识别装置包括光源生成模块28和光电感应器29；液体识别位点包括用于定位光源生成模块28的上位点和用于定位光电感应器29的下位点，上位点和下位点分别设于芯片主体的外侧，上位点、相应的进液口或出液口、下位点呈垂直线依次布设。相应地，光源生成模块28、相应的进液口或出液口、第一光电感应器29呈垂直线依次布设。由于液体识别装置可设于液体定量腔室或检测腔室的进液口或出液口处，因此此处的“相应的进液口或出液口”对应于液体定量腔室或检测腔室的进液口或出液口；例如，当液体定量腔室的出液口设置液体识别装置时，光源生成模块、液体定量腔室的出液口、光电感应器呈垂直线依次布设；当液体定量腔室的进液口设置液体识别装置时，光源生成模块、液体定量腔室的进液口、光电感应器呈垂直线依次布设；当样品定量腔室的出液口设置液体识别装置时，光源生成模块、样品定量腔室的出液口、光电感应器呈垂直线依次布设。
采用光学传感来对液体识别、定量和控制，相对于导电式的接触方式，此方法减少了金属对芯片内反应体系的干预，可提高检测效率，进而提高定量的准确性，同时这样的液体识别装置可设于微流控芯片外部，便于固定在仪器中，而无需设置在芯片上，降低了芯片的加工难度。使用时，只需将光源生成模块和光电感应器对准液体识别位点放置即可。具体地，芯片主体包括顶板1和底板20；顶板1与底板20层叠连接；顶板1与底板20的连接处设置有主流体通道和多个功能腔室；光源生成模块28设置定位在与液体定量腔室的进液口或出液口对应的顶板1的相应位置的正上方，光电感应器29定位在与液体定量腔室的进液口或出液口对应的底 板20的相应位置的正下方。
光源生成模块即能够提供光源的模块，其可为LED、卤素灯、激光灯等。在光源的照射下，由于气体、液体对光的的透射率和折射率不同，照射到光电感应器的光强不同，光电感应器便可以识别气体和液体，从而辨别液体是否到感应的点位。，当液体流至进液口或出液口时，液体识别装置可进行快速识别，从而控制液体驱动装置。
实施例6
本发明的实施例还提供了一种具有微流控芯片的分析仪器，其包括仪器框架、至少一个试剂存储池、液体驱动装置、检测装置和以上任一实施例中的微流控芯片；其中，微流控芯片安装在所述仪器框架中；液体驱动装置与微流控芯片的液体驱动力入口相连；试剂存储池与对应的试剂入口可通断地连通；检测装置用于接收处理微流控芯片发出的检测信号。
可选地，所述液体驱动装置为柱塞泵；每个所述试剂存储池上均设有与外界空气连通的开口。
以上所述是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也视为本发明的保护范围。

Claims (14)

  1. 一种微流控芯片，其特征在于，包括芯片主体、以及设置在所述芯片主体上的进样口、液体驱动力入口、主流体通道和多个功能腔室；
    所述主流体通道连通所述多个功能腔室，所述进样口和所述液体驱动力入口分别与所述主流体通道连通，所述液体驱动力入口用于连接液体驱动装置以驱动液体流入或流出功能腔室；
    所述多个功能腔室中的至少一个为液体定量腔室；所述液体定量腔室具有预定的容积，且在液体定量腔室的出液口处设置有液体识别位点，需定量的液体从所述液体定量腔室的进液口流入所述液体定量腔室，充满所述液体定量腔室后到达所述出液口。
  2. 根据权利要求1所述的微流控芯片，其特征在于，所述液体定量腔室包括试剂定量腔室，所述试剂定量腔室的进液口与试剂支通道的一端连通，所述试剂支通道的另一端与试剂入口连通。
  3. 根据权利要求1所述的微流控芯片，其特征在于，所述液体定量腔室的进液口处也设置有液体识别位点。
  4. 根据权利要求1所述的微流控芯片，其特征在于，所述液体驱动装置为柱塞泵。
  5. 根据权利要求1所述的微流控芯片，其特征在于，所述功能腔室包括检测腔室，所述检测腔室具有预定的容积，且在所述检测腔室的出液口处设置有液体识别位点，待检测的液体经所述检测腔室的进液口流入所述检测腔室，充满所述检测腔室后到达出液口。
  6. 根据权利要求5所述的微流控芯片，其特征在于，所述检测腔室的进液口处也设置有液体识别位点。
  7. 根据权利要求1～6中任一项所述的微流控芯片，其特征在于，所述液体识别位点用于 定位液体识别装置；所述液体识别装置包括光源生成模块和光电感应器；
    所述液体识别位点包括用于定位所述光源生成模块的上位点和用于定位所述光电感应器的下位点，所述上位点和所述下位点分别设于所述芯片主体的外侧，所述上位点和下位点的位置与相应的出液口或进液口对应，以使得定位后的所述光源生成模块、相应的出液口或进液口、所述光电感应器呈垂直线依次布设。
  8. 根据权利要求1所述的微流控芯片，其特征在于，所述液体定量腔室为六边形结构的腔室。
  9. 根据权利要求1所述的微流控芯片，其特征在于，所述液体定量腔室的进液口的宽度为0.3-3mm，高度为0.3-3mm；所述液体定量腔室的出液口的宽度为0.3-3mm，高度为0.3-3mm；或
    所述液体定量腔室的表面为经亲水性表面修饰后而形成的表面；所述液体定量腔室的进液口的宽度为0.3-5mm，高度为0.3-3mm；所述液体定量腔室的出液口的宽度为0.3-5mm，高度为0.3-3mm；或
    所述液体定量腔室的表面为经疏水性表面修饰后而形成的表面，所述液体定量腔室的进液口的宽度为0.3-2mm，高度为0.3-3mm；所述液体定量腔室的出液口的宽度为0.3-2mm，高度为0.3-3mm。
  10. 根据权利要求1所述的微流控芯片，其特征在于，所述芯片主体包括顶板和底板；所述顶板与所述底板层叠连接，所述顶板与所述底板的连接处设置有所述主流体通道和所述多个功能腔室。
  11. 根据权利要求10所述的微流控芯片，其特征在于，所述底板为光滑的平板，所述顶板上设置有微孔、微通道或微腔体以与所述底板形成所述进样口、液体驱动力入口、主流体通道或功能腔室。
  12. 根据权利要求1所述的微流控芯片，其特征在于，所述进样口和所述液体驱动力入口分别设置在所述主流体通道的两端。
  13. 一种具有微流控芯片的分析仪器，其特征在于，包括仪器框架、至少一个试剂存储池、液体驱动装置、检测装置和权利要求1～12中任一项所述的微流控芯片；其中，所述微流控芯片安装在所述仪器框架中；所述液体驱动装置与微流控芯片的液体驱动力入口相连；所述试剂存储池与对应的试剂入口可通断地连通；所述检测装置用于接收处理微流控芯片发出的检测信号。
  14. 根据权利要求13所述的具有微流控芯片的分析仪器，其特征在于，所述液体驱动装置为柱塞泵；每个所述试剂存储池上均设有与外界空气连通的开口。
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