CN108761055A - A kind of micro-fluidic chip and the analytical instrument with the micro-fluidic chip - Google Patents
A kind of micro-fluidic chip and the analytical instrument with the micro-fluidic chip Download PDFInfo
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- CN108761055A CN108761055A CN201810394846.6A CN201810394846A CN108761055A CN 108761055 A CN108761055 A CN 108761055A CN 201810394846 A CN201810394846 A CN 201810394846A CN 108761055 A CN108761055 A CN 108761055A
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Classifications
-
- 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
-
- 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
Abstract
The invention discloses a kind of micro-fluidic chips comprising chip body and the injection port being arranged in the chip body, liquid driven power entrance, main fluid passageway and multiple function chambers.The micro-fluidic chip of the present invention realizes that the identification of liquid is positioned and quantified by specific liquid quantitative chamber, reduces the manufacture craft difficulty of chip, improves quantitative accuracy.
Description
Technical field
The present invention relates to medical instruments field more particularly to a kind of micro-fluidic chip and with the analysis of the micro-fluidic chip
Instrument.
Background technology
In-vitro diagnosis (In Vitro Diagnosis, IVD) refers to taking-up sample (blood, body fluid, the tissue from human body
Deng) be detected analysis to being diagnosed to disease, need corresponding instrument and reagent in detection process, and these instruments and
Reagent just constitutes extracorporeal diagnostic system.The system of in-vitro diagnosis is roughly divided into two kinds；One is be with inspection center laboratory
Represent, it have system modular, automation, the carry out Sample of pipeline system, to also have high throughput, high efficiency,
The advantage of high sensitive, but whole system also has somewhat expensive, shared volume is big, the defect for needing professional to operate, it
It is mainly used in large hospital.Another be with detect immediately (point-of-care testing, POCT) be representative, it
System have it is integrated, miniaturization, carry out Sample whenever and wherever possible, to also have price material benefit, it is easy to operate, as a result
Report timely advantage, but its test result compares that there is also sensitivity, the not high disadvantages of stability with central laboratory.
For POCT, has both at home and abroad and microflow control technique is applied in the product of in-vitro diagnosis.It is micro-fluidic
(microfluidics) it is a cross discipline for carrying out control operation on the chip with microchannel to microfluid at one piece,
It is related to the fields such as biology, chemistry, fluid physics, electronics, optics, mechanical engineering.Micro fluidic device is commonly known as micro-fluidic
Chip, also referred to as chip lab (Lab on a Chip).Usually the sample system of biology, chemistry, medical analysis process
The basic operations such as standby, reaction, separation, detection are concentrated on one chip, and a system function is completed.Existing micro-fluidic chip
Mainly based on qualitative detection, the micro-fluidic chip quantitatively detected is less, and existing quantitative micro-fluidic chip prepares complicated, life
Production efficiency is low, and as the Chinese patent application of Publication No. " CN105214744 " discloses, " a kind of magnetic microparticle chemiluminescence is micro-fluidic
Chip ", the micro-fluidic chip include top plate and bottom plate, wherein the top plate includes air pump, adding mouth, sample fill area, mark
Remember ligand storage pool and sample mixed zone；The bottom plate includes filtering area, magnetic particle coating area, cleaning area, detection zone, cleaning solution
Storage pool, luminous substrate liquid storage pool and liquid release channel；The top plate and bottom plate all include liquid sensing device, are used for true
Determine the flow regime of liquid in micro-fluidic chip and whether be mixed into bubble etc., the chip in this patent uses multilayered structure, and
It realizes quantifying for liquid using the bag container of specific volume, although such D-M (Determiner-Measure) construction is simple, bag container surface is easily
Liquid hanging bag phenomenon occur, (when extruding liquid from bag container, partially liq extension stays in bag, can not ensure liquid is complete
Portion extrudes), and the deflection that bag container is squeezed every time is all different, so each amount of liquid remained in bag container is not
Unanimously, and then the amount of liquid that is extruded of liquid is different, and when in particular for gobbet, the error of bag container is more
Greatly, relative to micro-fluidic chip, what is needed is all tens microlitres of amount, so the quantitative accurate degree of bag container is unable to reach and wants
It asks, dosing accuracy is poor, and to influence testing result, while bag container needs to be built into chip, increases the life of chip
Produce difficulty.
Invention content
To solve the above problems, on the one hand the present invention provides a kind of micro-fluidic chip, quantitative detection may be implemented, and
It is simple in structure, reduce the manufacture craft difficulty of chip.
The technical solution adopted by the present invention is：A kind of micro-fluidic chip, including chip body and setting are in the chip
Injection port, liquid driven power entrance, main fluid passageway in main body and multiple function chambers；
The main fluid passageway is connected to the multiple function chamber, the injection port and liquid driven power entrance difference
It is connected to the main fluid passageway, the liquid driven power entrance is for connecting liquid driving device to drive liquid to flow into or flow
Go out function chamber；
At least one of the multiple function chamber is liquid quantitative chamber；The liquid quantitative chamber has scheduled
Volume, and Liquid identification site is provided at the liquid outlet of liquid quantitative chamber, the liquid that need to be quantified is from the liquid quantitative
The inlet of chamber flows into the liquid quantitative chamber, full of reaching the liquid outlet after the liquid quantitative chamber.
The liquid quantitative chamber includes reagent quantitative chamber in one of the embodiments, the reagent quantitative chamber
Inlet be connected to one end of reagent subchannel, the other end of the reagent subchannel is connected to reagent inlet.
In one of the embodiments, Liquid identification site is also equipped at the inlet of the liquid quantitative chamber.
The liquid driving device is plunger pump in one of the embodiments,.
The liquid quantitative chamber further includes sample amounts chamber in one of the embodiments, described in fluid sample warp
Injection port flows into the sample amounts chamber and is quantified；The sample amounts chamber is located at the upper of the reagent quantitative chamber
Trip；
The air subchannel for being additionally provided with air intake on the micro-fluidic chip and communicating therewith, the air subchannel
One end is connected to the air intake, and the main fluid passageway between the other end and the sample amounts chamber and the injection port connects
Logical, the other end of the air subchannel is with the connectivity part of the main fluid passageway adjacent to the sample amounts chamber.
The function chamber includes detection chambers in one of the embodiments, and the detection chambers have scheduled appearance
Product, and Liquid identification site is provided at the liquid outlet of the detection chambers, liquid to be detected is through the detection chambers
Inlet flows into the detection chambers, full of reaching liquid outlet after the detection chambers.
In one of the embodiments, Liquid identification site is also equipped at the inlet of the detection chambers.
The Liquid identification site is for positioning Liquid identification device in one of the embodiments,；The Liquid identification
Device includes light source generation module and photoelectric sensor；
The Liquid identification site includes upper site for positioning the light source generation module and for positioning the light
The lower site of electric inductor, the upper site and the lower site are respectively arranged on the outside of the chip body, the upper site
It is corresponding with the position in lower site and corresponding liquid outlet or inlet, so that the light source generation module after positioning, corresponding
Liquid outlet or inlet, the perpendicular line of the photoelectric sensor lay successively.
The liquid quantitative chamber is the chamber of hexagonal structure in one of the embodiments,.
The width of the inlet of the liquid quantitative chamber is 0.3-3mm in one of the embodiments, is highly 0.3-
3mm；The width of the liquid outlet of the liquid quantitative chamber is 0.3-3mm, is highly 0.3-3mm；Or
The surface of the liquid quantitative chamber is the surface formed after hydrophilic surface is modified；The liquid quantitative chamber
The width of the inlet of room is 0.3-5mm, is highly 0.3-3mm；The width of the liquid outlet of the liquid quantitative chamber is 0.3-
5mm is highly 0.3-3mm；Or
The surface of the liquid quantitative chamber is the surface formed after hydrophobic surface is modified, the liquid quantitative chamber
The width of the inlet of room is 0.3-2mm, is highly 0.3-3mm；The width of the liquid outlet of the liquid quantitative chamber is 0.3-
2mm is highly 0.3-3mm.
The chip body includes top plate and bottom plate in one of the embodiments,；The top plate is laminated with the bottom plate
The junction of connection, the top plate and the bottom plate is provided with the main fluid passageway and the multiple function chamber.
The bottom plate is smooth tablet in one of the embodiments, be provided on the top plate micropore, microchannel or
Microcavity body with the bottom plate to form the injection port, liquid driven power entrance, main fluid passageway or function chamber.
The injection port and the liquid driven power entrance are separately positioned on the main fluid in one of the embodiments,
The both ends in channel.
On the other hand, the present invention also provides a kind of analytical instrument with micro-fluidic chip comprising apparatus frame, extremely
A few reagent storage pool, liquid driving device, detection device and above-described micro-fluidic chip；Wherein, described micro-fluidic
Chip is mounted in the apparatus frame；The liquid driving device is connected with the liquid driven power entrance of micro-fluidic chip；Institute
Reagent storage pool is stated can be connected to break-make with corresponding reagent inlet；The detection device is for receiving processing micro-fluidic chip hair
The detection signal gone out.
The liquid driving device is plunger pump in one of the embodiments,；It is all provided on each reagent storage pool
There is the opening being connected to outside air.
Compared with the existing technology, the present invention has the advantages that：
Micro-fluidic chip provided by the invention realizes liquid by specific liquid quantitative chamber combination liquid driving device
Body quantifies, and is embedded in chip reagent packet by squeezing relative to existing and realizes that quantitative technology, liquid of the invention are fixed
Chamber is measured, quantitative accuracy is improved；And reagent is external to chip, compared with the existing technology in multilayer chiop combine and
Reagent plates in chip, reduces the manufacture craft difficulty of chip, improves the accuracy of detection.
The chip body of micro-fluidic chip of the present invention may include the top plate and bottom plate being stacked, and need to machine
On the top plate that structure can be arranged, bottom plate is only smooth tablet, can further decrease the manufacture craft difficulty of chip in this way, carry
High efficiency.
Description of the drawings
Fig. 1 is a kind of structural schematic diagram of embodiment of micro-fluidic chip provided by the invention；
Fig. 2 is the schematic cross-section of Liquid identification device provided by the invention；
Fig. 3 is a kind of arrangement structure for sensor figure of embodiment of micro-fluidic chip provided by the invention；
Fig. 4 is the schematic cross-section of magnet installation position when micro-fluidic chip provided by the invention uses；
Fig. 5 is a kind of structural schematic diagram of embodiment of liquid driving device provided by the invention；
Its wherein, 1, top plate；2, injection port, 3, Whole Blood Filtration area；4, sample amounts area；5, enzyme mark primary antibody embeds area；6,
First mixing channel；7, magnetic mark secondary antibody embeds area；8, the second mixing channel；9, chemiluminescence detection area；10, dilution entrance；
11, substrate luminescent solution entrance；12, filter washing water inlet；13, liquid driven power entrance；14, air intake；15, gasket；16, dilute
Release liquid subchannel；17, substrate luminescent solution subchannel；18, cleaning solution subchannel；19, plunger pump；20, bottom plate；21, dilution is deposited
Reservoir；22, substrate luminescent solution storage pool；23, cleaning solution storage pool；24, waste liquid pool；25a/25b, magnet；26, magnetic bead；27, empty
Gas subchannel；28, light source generation module；29, photoelectric sensor；191, the inlet of plunger pump；192, the liquid outlet of plunger pump；
193, plunger；194, pump chamber.
Specific implementation mode
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation describes, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art are obtained every other without creative efforts
Embodiment shall fall within the protection scope of the present invention.
Embodiment 1
A kind of micro-fluidic chip is present embodiments provided, which includes chip body and be arranged described
Injection port, liquid driven power entrance, main fluid passageway in chip body and multiple function chambers；It is specifically described below.
In the present embodiment, main fluid passageway is connected to multiple function chambers, to guide stream of the fluid between function chamber
It is dynamic.
In the present embodiment, function chamber at least has the function of accommodating, it is preferable that function chamber accommodates function except tool
Outside, also have the function of other, other functions can be realized in functional chamber room, can also be that function chamber combines it external
Necessary parts (these necessary components can fix in chip exterior, without setting in the chip or its surface) realize jointly.
In the present embodiment, injection port and liquid driven power entrance are connected to main fluid passageway respectively, and liquid driven power enters
For connecting liquid driving device, to drive, liquid flows into mouth or downstream chamber, injection port are used to fluid sample introducing master
In fluid channel, fluid sample enters each function chamber through main fluid passageway.
In the present embodiment, at least one of multiple function chambers are liquid quantitative chamber；The liquid quantitative chamber
With scheduled volume, and Liquid identification site is provided at the liquid outlet of liquid quantitative chamber, the liquid that need to be quantified is from institute
The inlet for stating liquid quantitative chamber flows into the liquid quantitative chamber, full of going out liquid described in being reached after the liquid quantitative chamber
Mouthful.Liquid identification site is for positioning or fixing liquid identification device, Liquid identification device liquid for identification, when liquid is flow to
When at Liquid identification site, Liquid identification device can recognize that liquid and provide liquid arriving signal；Therefore when liquid reaches out liquid
When at mouthful, Liquid identification device can provide liquid arriving signal, and liquid quantitative chamber is full of by indicating liquid, controls liquid at this time
Body drive stops driving liquid, realizes liquid quantifying in liquid quantitative chamber.
Micro-fluidic chip provided in this embodiment is realized by specific liquid quantitative chamber combination liquid driving device
Liquid quantifies, and is embedded in chip reagent packet by squeezing relative to existing and realizes quantitative technology, liquid of the invention
Quantitative chamber, improves quantitative accuracy.Reagent is external to chip, and middle multilayer chiop is combined and tried compared with the existing technology
Agent plates in chip, reduces the manufacture craft difficulty of chip, improves the accuracy of detection.
It should be noted that main fluid passageway and multiple function chambers can pass through laser processing, model injection molding etc.
Various ways shape inside chip body, also can by being set as the top plate and bottom plate of separate type, on top plate or bottom plate plus
Work goes out specific shape, is then mutually packaged together；Since former processing method is relatively complicated, in a preferred embodiment
In, the chip body includes top plate and bottom plate；Top plate is connected with bottom plate stacking；The junction of top plate and bottom plate is provided with described
Main fluid passageway and multiple function chambers；It is highly preferred that bottom plate is smooth tablet, setting micropore, microchannel or micro- on top plate
Cavity with bottom plate to cooperatively form injection port, liquid driven power entrance, main fluid passageway or multiple function chambers, such miniflow
Control chip prepare get up it is more convenient, reduce producting process difficulty, only required specific structure need to be processed on top plate,
Improve production efficiency.Specifically, the bottom plate be smooth tablet, the top plate be equipped with multiple microchannels with the bottom
Hardened conjunction forms main fluid passageway, and the top plate is equipped with multiple microcavitys to be combined to form the multiple functional chamber with the bottom plate
Room, the top plate are equipped with bottom plate described in multiple Kong Yiyu in conjunction with the formation injection port or liquid driven power entrance；In order to just
It is typically larger than the size of other entrances in the size of sample introduction, injection port.
The quantity of liquid quantitative chamber, (such as fluid sample, reaction reagent, the sample treatment examination of its quantitative kind of liquid
Agent etc.), the types of installation position and other function chambers can be selected according to actual needs.
Preferably, liquid quantitative chamber includes reagent quantitative chamber, inlet and the reagent subchannel of reagent quantitative chamber
One end connection, the other end of reagent subchannel is connected to reagent inlet, and reagent enters reagent through reagent inlet, reagent subchannel
Quantitative chamber is quantified.When carrying out quantitative detection using micro-fluidic chip, fluid sample (is flowed into from injection port to be measured
Liquid) and the amount of reagent (such as reaction reagent, sample processing reagent etc.) quantified, the quantitative needs of usual reagent exist
Chip interior is completed, and fluid sample selectively realizes outside micro-fluidic chip and quantify that reagent quantitative chamber is in the present embodiment
For quantifying for reagent." liquid quantitative chamber includes reagent quantitative chamber " herein is interpreted as liquid quantitative chamber should be at least
With for this type of the reagent quantitative chamber of quantitative reagent, naturally it is also possible to further comprise for other quantitative liquid types
The liquid quantitative chamber of type, such as the sample amounts chamber for quantitative liquid sample.
The quantity of reagent quantitative chamber can be one, it is two or more, can be carried out according to the actual needs of micro-fluidic chip
Selection.Such as when micro-fluidic chip is chemiluminescence micro-fluidic chip, in order to realize that chemiluminescence quantitatively detects, reagent quantitative
Chamber should at least be arranged one for quantifying substances luminescent solution, remaining reacts necessary substance such as enzyme mark primary antibody, magnetic mark two
It is anti-to be embedded in respectively in enzyme mark primary antibody embedding chamber and the two functional chamber rooms of magnetic mark secondary antibody embedding chamber；Preferably, magnetic mark
It is reagent quantitative chamber that secondary antibody, which embeds chamber, is inside not only embedded with magnetic mark secondary antibody, it may also be used for quantifying substances luminescent solution；It is more excellent
Selection of land, magnetic mark secondary antibody embedding chamber are additionally operable to magnetic bead cleaning.
Optionally, reagent inlet can be connect to break-make with reagent storage pool by valve, and reagent storage pool is equipped with and the external world
The opening of air communication, reagent storage pool is equipped with opening can will be in the liquid drain to chip in it convenient for liquid driving device；
For the ease of the preparation of chip, it is preferable that reagent storage pool is set to the outside of micro-fluidic chip, and when use pacifies reagent storage pool
At reagent inlet, to introduce the reagents into chip.
Optionally, it is also equipped with Liquid identification site at the inlet of liquid quantitative chamber.This Liquid identification site is same
It can be used for positioning or fix liquid identification device, setting can be convenient for being monitored the flowing of liquid in chip, controlling, also
The mixing between two kinds of quantitative liquids, such as the mixing between fluid sample and reagent can be achieved.Portion in the chip, to realize
The mixing of two kinds of liquid needs two kinds of liquid contacts, and gap is not present in centre, and the micro-fluidic chip of the present invention is to real simultaneously
Quantitative and two kinds of liquid the contact of existing liquid, this requires the liquid after one of which is quantitative to rest on pre-position,
Another liquid preferably begins to flow into liquid quantitative chamber from this precalculated position, realizes and quantifies in liquid quantitative chamber, and this
At the inlet of the optimal selection in precalculated position, that is, liquid quantitative chamber；Liquid identification site is set at inlet to position liquid
Body identification device, this Liquid identification device can provide the stop indication signal of one of which liquid and the feed liquor of another liquid
Signal, under the cooperation of the Liquid identification device at the liquid outlet of liquid quantitative chamber, you can realize liquid quantitative and two
The contact of kind liquid.Next, illustrating one of lower micro-fluidic chip by taking the mixing of fluid sample and reagent as an example makes
Use method：
Micro-fluidic chip is in use, reagent inlet can be connect to break-make with reagent storage pool by valve, reagent storage pool
Equipped with the opening being connected to outside air.Fluid sample is (for the ease of accurately mixing or reacting, fluid sample with quantitative reagent
The preferably fluid sample of predetermined amount) under the action of liquid driving device from injection port at through main fluid passageway to flow to reagent fixed
When measuring at the inlet of chamber, the Liquid identification device being positioned on the Liquid identification site of reagent quantitative chamber obtains signal,
It controls liquid driving device and stops driving effect, fluid sample stops flowing at this time, and the front end of fluid sample rests on inlet
Place；Then, air intake is closed, opens the valve between reagent inlet and reagent storage pool, reagent is in liquid driving device
Enter in reagent quantitative chamber from inlet of the reagent inlet through reagent subchannel, reagent quantitative chamber under effect, is tried when flowing to
When agent quantifies the liquid outlet of chamber, reagent is full of reagent quantitative chamber, closes between reagent inlet and reagent storage pool at this time
Valve opens air intake, and the quantitative reagent of reagent quantitative chamber and fluid sample continue under the action of liquid driving device
Flowing, and can realize positive reverse movement under the positive pressure of liquid driving device, negative pressure alternating action and realize mixing and/or instead
It answers.
In order to reduce influence of the reagent subchannel for the Liquid identification device at reagent quantitative chamber inlet, while just
In processing and fabricating, specifically, the feed liquor that one end of the reagent subchannel passes through the main fluid passageway and reagent quantitative chamber
Mouthful connection, the connectivity part of the reagent subchannel and the main fluid passageway at the inlet of reagent quantitative chamber so that
It quantifies in controlled range, reduces quantitative error, for example, the connectivity part distance of the reagent subchannel and the main fluid passageway
The distance of the inlet of reagent quantitative chamber is 0.5~10mm (preferably 0.5-2mm).
Optionally, the injection port and the liquid driven power entrance are separately positioned on the both ends of the main fluid passageway.
Optionally, the function chamber includes detection chambers, and detection chambers have scheduled volume, and in detection chambers
It is provided with Liquid identification site at liquid outlet, liquid to be detected flows into detection chambers, full of reaching liquid outlet after detection chambers.
The Liquid identification site being arranged at the liquid outlet of detection chambers can be used for positioning or fix liquid identification device, when liquid to be detected
When body is reached at the liquid outlet of detection chambers, Liquid identification device sends out signal, and liquid driving device controls liquid to be detected
Stop flowing, can be detected at this time.Further, it also is provided with Liquid identification site at the inlet of detection chambers.
Optionally, liquid quantitative chamber still further comprises sample amounts chamber, and fluid sample flows into sample through injection port
Quantitative chamber is quantified；Sample amounts chamber is located at the upstream of reagent quantitative chamber；Air is additionally provided on micro-fluidic chip to enter
Mouthful and the air subchannel that communicates therewith, one end of the air subchannel be connected to the air intake, the other end with it is described
Main fluid passageway connection between sample amounts chamber and the injection port, the other end and the mainstream of the air subchannel
The connectivity part in body channel is adjacent to the sample amounts chamber；" neighbouring " usually can be regarded as " apart from sample amounts chamber herein
0.5~10mm of inlet (preferably 0.5-2mm) ".
Micro-fluidic chip in use, air intake and the air pipeline of chip exterior can be connected to break-make by valve,
Enter chip interior to control air.Fluid sample is under the action of liquid driving device through injection port from sample amounts chamber
Inlet flows into sample amounts chamber, when fluid sample is flow at the liquid outlet of sample amounts chamber, i.e., fixed full of sample
Chamber is measured, the Liquid identification device positioned on the Liquid identification site of liquid outlet is now placed in and sends out indication signal, control air
Entrance is opened, and since driving force needed for the flowing of air in air subchannel is small, and driving force needed for the flowing of fluid sample is more
Greatly, therefore fluid sample rests on air subchannel and the connectivity part of main fluid passageway does not continue to flow into sample amounts chamber,
Fluid sample quantifying in sample amounts chamber can be completed.Fluid sample after quantitative can be in the effect of liquid driving device
Lower to continue to flow at the inlet of reagent quantitative chamber, candidate agent quantifies quantitative rear (the process institute as above that chamber completes reagent
State), fluid sample and reagent realize mixing and/or reaction under the positive pressure of liquid driving device, negative pressure alternating action.
Micro-fluidic chip in the present embodiment can be convenient for quantifying for fluid sample due to being provided with sample amounts chamber, and
Without separately quantitative outside chip so that chip uses more convenient.
Optionally, the fluid sample is whole blood, and whole blood mistake is equipped between the injection port and the sample amounts chamber
Filter chamber room is equipped with whole blood filter membrane in the Whole Blood Filtration chamber；When micro-fluidic chip is used for clinical diagnosis, whole blood is common
Detect sample, when detection usually require to carry out whole blood separation with by whole blood serum or blood plasma separate, then with reagent into
Row reaction；Whole Blood Filtration chamber is set in chip, is used convenient for detection, while compared to first quantifying whole blood, then carries out whole blood point
From mode, between injection port and the sample amounts chamber be equipped with Whole Blood Filtration chamber, can be straight by sample amounts chamber
The dosage of quantitative serum or blood plasma is connect, measurement result is more accurate.The material of the whole blood filter membrane can be glass fibre, cotton linter
Fiber, polyester fiber, fiber or blend fibre；Optionally, the thickness of the Whole Blood Filtration filter bed is 0.2-2.5mm；It is described complete
The adsorption rate that blood filters filter bed is 4-150s/4cm, water imbibition 30-250mg/cm2。
Optionally, the liquid outlet in Whole Blood Filtration area is triangle liquid outlet；Whole Blood Filtration area area is 30-300mm2, wide
Angle for 2-20mm, a length of 5-25mm, depth 0.3-3mm, front end triangle is 15-160 DEG C.
Embodiment 2
Please refer to Fig. 1~Fig. 5, present embodiments provide a kind of chemiluminescence micro-fluidic chip comprising chip body, with
And be arranged in chip body injection port 2, liquid driven power entrance 13, substrate luminescent solution entrance 11, filter washing water inlet 12, bottom
Object luminescent solution subchannel 17, cleaning solution subchannel 18, main fluid passageway and multiple function chambers；It is specifically described below.
In the present embodiment, main fluid passageway is connected to multiple function chambers, to guide stream of the fluid between function chamber
It is dynamic.
Function chamber includes embedding area 5, magnetic mark secondary antibody embedding 7 and of area by the enzyme mark primary antibody that main fluid passageway is sequentially communicated
Chemiluminescence detection area 9.
Wherein, enzyme mark primary antibody embedding area 5 is embedded with enzyme mark primary antibody；Magnetic mark secondary antibody embedding area 7 is embedded with magnetic mark secondary antibody；Magnetic mark
It is liquid quantitative chamber that secondary antibody, which embeds area 7,；Liquid quantitative chamber be used for quantitative liquid, wait for quantitative liquid (such as substrate shine
Liquid) enter liquid quantitative chamber after, quantitative (obtaining desired amount of liquid) can be realized in liquid quantitative chamber, with calmly
The fluid sample of amount or the reaction of other reaction reagents, to realize quantitative detection.
In the present embodiment, liquid quantitative chamber has scheduled volume, and is set at the liquid outlet of liquid quantitative chamber
It is equipped with Liquid identification site, the liquid that need to be quantified quantifies chamber from the inlet influent of liquid quantitative chamber, hydraulically full
Liquid outlet is reached after quantitative chamber；For positioning or fixing liquid identification device, Liquid identification device is used in Liquid identification site
Identify liquid.When liquid reaches at liquid outlet, Liquid identification device can provide liquid arriving signal, and indicating liquid is by liquid
Quantitative chamber is full of, and is controlled liquid driving device at this time and is stopped driving liquid, you can realizes liquid in liquid quantitative chamber
It is quantitative.Chemiluminescence micro-fluidic chip realizes determining for liquid by specific liquid quantitative chamber combination liquid driving device
Amount, can be improved quantitative accuracy.
In the present embodiment, chemiluminescence detection area 9 is for housing chemiluminescence reaction product, with external detection device
In conjunction with completion detection process.
Injection port 2 is connected to main fluid passageway respectively with liquid driven power entrance 13, and driving force entrance 13 is for connecting liquid
Body drive is to drive liquid inflow or downstream chamber；Injection port 2 is used to fluid sample being introduced into main fluid passageway,
Fluid sample enters each function chamber through main fluid passageway.
In the present embodiment, one end of substrate luminescent solution subchannel 17 is connected to substrate luminescent solution entrance 11, the other end
The inlet that area 7 is embedded with magnetic mark secondary antibody is connected to, and substrate luminescent solution is through substrate luminescent solution entrance 11, substrate luminescent solution subchannel 17
It is quantified into magnetic mark secondary antibody embedding area 7.
One end of cleaning solution subchannel 18 is connected to filter washing water inlet 12, the other end and magnetic mark secondary antibody embedding area 7 into
Liquid mouth is connected to, and the cleaned liquid entrance 12 of cleaning solution, cleaning solution subchannel 18 enter magnetic mark secondary antibody embedding area 7 and carry out magnetic bead cleaning.
The micro-fluidic chip of the present embodiment is in use, substrate luminescent solution entrance 11, filter washing water inlet 12 are sent out with substrate respectively
Light liquid storage pool 22, cleaning solution storage pool 23 can be connected to break-make by valve V2, V3, substrate luminescent solution storage pool 22, cleaning
The opening being connected to outside air is respectively equipped on liquid storage pool 23；Liquid driving device is mounted on liquid driven power entrance 13
Place, flows to liquid in driving chip；The outside in magnetic mark secondary antibody embedding area 7 is fixed with magnet (such as magnet 25a, 25b), with
Just magnetic bead 26 is fixed.It is liquid quantitative chamber that magnetic mark secondary antibody, which embeds area, can be used for quantifying substances luminescent solution, optionally, may be used also
It is further used for quantitative cleaning solution.
One working method of the micro-fluidic chip of the present embodiment is as follows：The fluid sample of predetermined amount is (such as through diluted
Serum afterwards or blood plasma) under the action of liquid driving device at injection port 2 through main fluid passageway flow to enzyme mark primary antibody embedding
Area 5, with the enzyme mark primary antibody hybrid reaction wherein embedded, the magnetic mark secondary antibody of reaction solution arrival thereafter embeds area 7, with the magnetic wherein embedded
It marks secondary antibody and continues hybrid reaction, form the reactant of double antibodies sandwich structure on magnetic bead, magnetic bead is by magnet adsorption, and reactant is in magnetic
Under the action of pearl stablize magnetic mark secondary antibody embedding area 7 in, and remaining reaction solution under the action of liquid driving device through liquid
Chip is discharged in driving force entrance 13；Then, the air inflow end mouth (such as sample inlet) on chip is closed, cleaning solution storage is opened
Valve V3 between pond 23 and filter washing water inlet 12, cleaning solution under the action of liquid driving device cleaned liquid subchannel 18 into
Enter magnetic mark secondary antibody and embeds area 7 to be cleaned to magnetic bead therein, when magnetic mark secondary antibody embedding area 7 completes to quantify cleaning solution
When, you can the valve V3 between cleaning solution storage pool 23 and filter washing water inlet 12 is closed, air inflow end mouth is opened, after cleaning
Chip is discharged through liquid driven power entrance 13 under the action of liquid driving device in liquid can be repeatedly in order to ensure cleaning performance
(magnetic bead cleaning way is not limited to mode described herein, can also be for example, by the moving magnet in cleaning solution for several times for cleaning
Mode realizes the cleaning of magnetic bead)；The air inflow end mouth (such as sample inlet) being then switched off on chip opens the storage of substrate luminescent solution
The valve V2 between pond 22 and substrate luminescent solution entrance 11 is deposited, substrate luminescent solution is sent out under the action of liquid driving device through substrate
Light liquid subchannel 19 enter magnetic mark secondary antibody embed area 7, when magnetic mark secondary antibody embedding area 7 complete to substrate luminescent solution it is quantitative when, close
The valve V2 between substrate luminescent solution storage pool 22 and substrate luminescent solution entrance 11 is closed, liquid driving device stops driving effect,
Substrate luminescent solution is no longer flow into magnetic mark secondary antibody embedding area 7, opens the air inflow end mouth (such as sample inlet) on chip, magnetic mark two
Resist the reactant of substrate luminescent solution and magnetic capture after quantifying to carry out luminescence-producing reaction, removes magnet, the embedding of magnetic mark secondary antibody later
Reaction solution in area 7 flows into chemiluminescence detection area 9 under the action of liquid driving device and is detected.
Above-mentioned chemiluminescence microfluidic chip structure is compact, such as magnetic mark secondary antibody embedding area is applied not only to embedding magnetic mark two
It is anti-, it can be additionally used in quantifying substances luminescent solution as liquid quantitative chamber, and liquid quantitative chamber need not be separately set again, magnetic mark
Secondary antibody embedding Qu Haike is further used as the area cleaned for magnetic bead, and magnetic bead cleaning area need not separately be arranged again, greatlys save
The volume of chip；Meanwhile reagent storage pool (such as substrate luminescent solution storage pool, cleaning solution storage pool) is external to chip,
Reagent plates in chip compared with the existing technology, reduces the manufacture craft difficulty of chip, improves the accuracy of detection.
It should be noted that main fluid passageway and multiple function chambers can pass through laser processing, model injection molding etc.
Various ways shape inside chip body, also can by being set as the top plate and bottom plate of separate type, on top plate or bottom plate plus
Work goes out specific shape, is then mutually packaged together；Since former processing method is relatively complicated, in a preferred embodiment
In, chip body includes top plate 1 and bottom plate 20；Top plate 1 is connected with the stacking of bottom plate 20；Top plate 1 and the junction of bottom plate 20 are arranged
There are main fluid passageway and multiple function chambers；It is highly preferred that bottom plate 20 is smooth tablet, micropore, microchannel is arranged in top plate 20
Or microcavity body with bottom plate to cooperatively form injection port 2, liquid driven power entrance 13, substrate luminescent solution entrance 11, filter washing water inlet
12, substrate luminescent solution subchannel 17, cleaning solution subchannel 18, main fluid passageway or multiple function chambers, such micro-fluidic core
Piece prepare get up it is more convenient, further reduced producting process difficulty, only required specific structure need to be processed on top plate i.e.
Can, further improve production efficiency.In one embodiment, bottom plate 20 is smooth tablet, and top plate 1 is equipped with multiple micro- logical
To be combined to form main fluid passageway with bottom plate 20, top plate 1 is equipped with multiple microcavitys to be combined to form multiple functions with bottom plate 20 in road
Chamber, top plate 1 form injection port 2, liquid driven power entrance 13, substrate luminescent solution equipped with the combination of multiple Kong Yiyu bottom plates 20 and enter
Mouth 11 and filter washing water inlet 12；For the ease of sample introduction, the size of injection port 2 is typically larger than the size of other entrances.
Therefore, the chip body of above-mentioned chemiluminescence micro-fluidic chip may include the top plate and bottom plate being stacked, and need
On the top plate that the structure machined can be arranged, bottom plate is only smooth tablet, can further decrease the making of chip in this way
Technology difficulty improves production efficiency.
Optionally, it is also equipped with Liquid identification site at the inlet of liquid quantitative chamber.This Liquid identification site is set
Setting can be convenient for being monitored the flowing of liquid in chip and bubble that may be present, controlling, and can also be achieved two kinds of quantitative liquid
Mixing between mixing between body, such as fluid sample and reagent (such as reaction reagent, sample processing reagent).Further
Ground, it is liquid quantitative chamber that enzyme mark primary antibody, which embeds area 5 also, and dilution entrance 10 and dilution subchannel are additionally provided in chip body
16；One end of dilution subchannel 16 is connected to dilution entrance 10, and the other end and the inlet in enzyme mark primary antibody embedding area 5 connect
Logical, sample diluting liquid enters enzyme mark primary antibody embedding area 5 through dilution entrance, dilution subchannel and is quantified.Further,
It is respectively arranged with Liquid identification site at the inlet and liquid outlet in enzyme mark primary antibody embedding area 5, the liquid that need to be quantified is from its feed liquor
Mouth flows into enzyme mark primary antibody and embeds area 5, and liquid outlet is reached after embedding area 5 full of enzyme mark primary antibody.Sample diluting liquid can not only dilute
Fluid sample (such as serum, blood plasma), reduces its concentration and viscosity, wherein the substance contained can also reduce the sheet of fluid sample
Floors so that detection is more accurate, while sample diluting liquid can preferably redissolve enzyme mark primary antibody；In this technical solution, enzyme
Mark primary antibody embedding area can be used for quantifying sample diluting liquid, quantitative without realizing quantifying for sample diluting liquid in chip exterior
Sample diluting liquid can embed area in enzyme mark primary antibody and be mixed with quantitative fluid sample, can save manpower, and operation is more convenient.
In use, dilution entrance 10 can be connect to break-make with dilution storage pool 21 by valve V1, on dilution storage pool 21
Equipped with the opening being connected to outside air；The fluid sample (serum such as after diluted or blood plasma) of predetermined amount is in liquid
Enzyme mark primary antibody is flow to through main fluid passageway to embed at the inlet in area 5, close chip under the action of driving device from injection port 2
On air intake (such as sample inlet), open the valve V1 between dilution storage pool 21 and dilution entrance 10, sample is dilute
It releases liquid and enters enzyme mark primary antibody embedding area 5 through dilution subchannel 16 under the action of liquid driving device, when it is full of enzyme mark one
Anti- embedding area 5, when reaching at the liquid outlet in enzyme mark primary antibody embedding area 5, close dilution storage pool 21 and dilution entrance 10 it
Between valve V1, open air inflow end mouth (such as sample inlet), fluid sample and sample diluting liquid can be filled in liquid driven
Continue to flow under the suction function set, and can be in the positive pressure of liquid driving device, negative pressure alternating action in main fluid passageway, enzyme mark
Primary antibody, which embeds, realizes mixing in area 5, can also realize preferably mixing by the hybrid channel of setting certainly.
Optionally, chemiluminescence detection area 9 has scheduled volume, and in the inlet in chemiluminescence detection area 9 and goes out
Liquid identification site is respectively arranged at liquid mouth, the inlet of liquid to be detected through chemiluminescence detection area 9 flows into chemistry hair
Light detection area 9, full of liquid outlet is reached behind chemiluminescence detection area 9, the volume in chemiluminescence detection area 9 is less than or equal to magnetic mark two
The volume in anti-embedding area 7.The Liquid identification site being arranged at the liquid outlet in chemiluminescence detection area 9 can be used for positioning or fixer
Body identification device, reaction solution after substrate luminescent solution is reacted with the reactant of magnetic capture reach chemiluminescence detection area
Liquid outlet at when, Liquid identification device sends out signal, and liquid driving device controls reaction solution and stops flowing, can carry out at this time
Detection.
Optionally, for the ease of the mixing between fluid sample, reagent (sample diluting liquid, substrate luminescent solution etc.), mainstream
Body channel includes the first mixing channel 6 and the second mixing channel 8；First mixing channel 6 is set to enzyme mark primary antibody embedding area 5 and magnetic mark
Secondary antibody embeds between area 7；Second mixing channel 8 is set between magnetic mark secondary antibody embedding area 7 and chemiluminescence detection area 9.
Optionally, injection port 2 is separately positioned on the both ends of main fluid passageway with liquid driven power entrance 13.
As shown in figure 4, optionally, for the ease of fixed magnetic bead, chip body position corresponding with magnetic mark secondary antibody embedding area 7
Place is provided with magnet and fixes site；Further, it is carried out since the cleaning of magnetic bead can embed area 7 in magnetic mark secondary antibody, in order to more preferable
Realize that magnetic bead cleaning, the embedding of magnetic mark secondary antibody respectively lay the magnet for phase magnet 25a, 25b above and below area 7 in ground
Fixed site, two magnet 25a, the diagonally opposing corner that 25b corresponds to magnetic mark secondary antibody embedding area 7 are laid.
Optionally, liquid driving device is plunger pump 19, and description is suitable for this implementation as described in plunger pump in embodiment 3
Example.
Optionally, function chamber further includes sample amounts chamber 4, and sample amounts chamber 4 is also liquid quantitative chamber, liquid
Sample flows into sample amounts chamber 4 through injection port and is quantified；Sample amounts chamber 4 is located at the upstream in enzyme mark primary antibody embedding area 5；
The air subchannel 27 for being additionally provided with air intake 14 on micro-fluidic chip and communicating therewith, one end of air subchannel 27 and air
Entrance 14 is connected to, and the main fluid passageway between the other end and sample amounts chamber 4 and injection port 2 is connected to, air subchannel 27
The connectivity part of the other end and main fluid passageway is adjacent to sample amounts chamber 4." neighbouring " usually can be regarded as " fixed apart from sample herein
Measure the 1~10mm " of inlet of chamber 4.By being provided with sample amounts chamber, fluid sample can be convenient for quantify, without
It is separately quantitative outside chip so that chip uses more convenient.Further, it is provided with liquid at the liquid outlet of sample amounts chamber 4
Body recognition site, the liquid that need to be quantified flow into sample amounts chamber 4 from its inlet, full of being reached out after sample amounts chamber 4
Liquid mouth.Further, it is also equipped with Liquid identification site at the inlet of sample amounts chamber 4.
Micro-fluidic chip in use, air intake and the air pipeline of chip exterior can be connected to break-make by valve,
Enter chip interior to control air.Fluid sample is under the action of liquid driving device through injection port from sample amounts chamber
Inlet flows into sample amounts chamber, when fluid sample is flow at the liquid outlet of sample amounts chamber, i.e., fixed full of sample
Chamber is measured, the Liquid identification device positioned on the Liquid identification site of liquid outlet is now placed in and sends out indication signal, control air
Entrance is opened, since driving pressure needed for the flowing of air in air subchannel is small, and driven needed for the flowing of fluid sample
Pressure bigger, therefore fluid sample rests on air subchannel and the connectivity part of main fluid passageway does not continue to flow into sample amounts
Chamber, you can complete fluid sample quantifying in sample amounts chamber.Fluid sample after quantitative can be in liquid driving device
Under the action of continue to flow to enzyme mark primary antibody embedding area.
Optionally, fluid sample is whole blood, and Whole Blood Filtration area 3, whole blood are equipped between injection port 7 and sample amounts chamber 4
Whole blood filter membrane is equipped in filtering area 3；When micro-fluidic chip is used for clinical diagnosis, whole blood is common detection sample, when detection
It is generally necessary to carry out whole blood separation with by whole blood serum or blood plasma separate, then reacted with reagent；It is set in chip
Whole Blood Filtration area is set, is used convenient for detection, while compared to whole blood is first quantified, then the mode of whole blood separation is carried out, in injection port
Between sample amounts chamber be equipped with Whole Blood Filtration area, can by the dosage of sample amounts chamber direct quantitative serum or blood plasma,
Measurement result is more accurate.The material of whole blood filter membrane can be glass fibre, cotton linter fiber, polyester fiber, fiber or blended fibre
Dimension；Optionally, the thickness of Whole Blood Filtration filter bed is 0.2-2.5mm；The adsorption rate of Whole Blood Filtration filter bed is 4-150s/4cm, is inhaled
Aqueous is 30-250mg/cm2。
The description as described in liquid quantitative chamber is suitable for above-described liquid quantitative chamber (including magnetic mark in embodiment 4
Secondary antibody embed area 7, enzyme mark primary antibody embedding area 5 and sample amounts chamber 4) description, details are not described herein.
Description is suitable for above-described Liquid identification as described in Liquid identification site and Liquid identification device in embodiment 5
The description of site and Liquid identification device, details are not described herein.
Optionally, the connectivity part position of the other end of substrate luminescent solution subchannel 17 and the inlet in magnetic mark secondary antibody embedding area 7
In the main fluid passageway for embedding the inlet in area 7 in magnetic mark secondary antibody；In one embodiment, " neighbouring " is interpreted as " distance herein
Magnetic mark secondary antibody embeds the 0.5~10mm of inlet (preferably 0.5~2mm) in area 7 ".
Optionally, the cleaned liquid entrance 12 of cleaning solution, cleaning solution subchannel 18 enter magnetic mark secondary antibody embedding area 7 and are determined
Amount；The connectivity part of the other end of cleaning solution subchannel 18 and the inlet in magnetic mark secondary antibody embedding area 7 is located at neighbouring with inlet
In main fluid passageway；In one embodiment, herein " neighbouring " be interpreted as " apart from magnetic mark secondary antibody embed area 7 inlet 0.5~
10mm (preferably 0.5~2mm) ".Preferably, the inlet of the other end of cleaning solution subchannel 18 and magnetic mark secondary antibody embedding area 7
The other end and magnetic mark secondary antibody of the connectivity part in substrate luminescent solution subchannel 17 embed area 7 inlet connectivity part downstream,
It can avoid substrate luminescent solution in this way and be cleaned liquid dilution.
Optionally, the connectivity part of the inlet in the other end of dilution subchannel 16 and enzyme mark primary antibody embedding area 5 be located at
Enzyme mark primary antibody embeds in the neighbouring main fluid passageway of inlet in area 5；In one embodiment, herein " neighbouring " be interpreted as " away from
From 0.5~10mm of inlet (preferably 0.5~2mm) that enzyme mark primary antibody embeds area 5 ".
Optionally, the volume of injection port 2 is 5ul-300ul.
Optionally, the liquid outlet in Whole Blood Filtration area 3 is triangle liquid outlet；3 area of Whole Blood Filtration area is 30-300mm2,
Width is 2-20mm, a length of 5-25mm, depth 0.3-3mm, and the angle of front end triangle is 15-160 DEG C.
Optionally, the volume of sample amounts chamber 4 is 1-50ul.
Optionally, the volume in enzyme mark primary antibody embedding area 5 is 5-50ul.
Optionally, the width of the first mixing pipeline 6 and the second mixing pipeline 8 is 200-2000um, a length of 5mm-40mm, and depth is
0.2-3mm。
Optionally, the volume in magnetic mark secondary antibody embedding area 7 is 10-200ul.
Optionally, the volume in chemiluminescence detection area 9 is 10-200ul.
Next, in conjunction with Fig. 1~Fig. 5, a kind of detection side of the micro-fluidic chip of embodiment according to the present invention is described
Method.The method comprising the steps of 101 to step 110, and each step is specific as follows：
Step 101：Will respectively with dilution storage pool 21, substrate luminescent solution storage pool 22, cleaning solution storage pool 23, plunger
Pump 19, the draw point of air communication are inserted into the closed pad 15 in chip, wherein draw point respectively with dilution entrance 10, substrate luminescent solution
Entrance 11, filter washing water inlet 12, liquid driven power entrance 13, air intake 14 connect；Whole blood sample is added to injection port 2,
It opens solenoid valve V4 and negative-pressure sucking is generated by plunger pump 19, by whole blood sample sucking Whole Blood Filtration area 3.
Step 102：Whole blood sample completes filtered serum and is inhaled into sample amounts chamber 4, and by sample amounts chamber
The photoelectric sensor (a1, a2) being arranged on 4 inlet of room and liquid outlet completes the quantitative measurment of serum.
It is that inductor output voltage value changes when whole blood sample passes through above photoelectric sensor a1, gives system one
Identification signal judges the flow locations of liquid in the chips.When sample passes through photoelectric sensor a2, judgement sample determines sample
Amount chamber 4 is full of, and the intrinsic volume in the region is the quantitative values of sample.
Step 103：It blocks injection port 2 and opens solenoid valve V5 so that serum is inhaled into enzyme mark primary antibody embedding area 5.
Step 104：When the photoelectric sensor (b1) being arranged on the inlet that enzyme mark primary antibody embeds area 5 detects serum,
Solenoid valve V5 is closed, solenoid valve V1 is opened so that external sample dilution enters enzyme mark primary antibody from solenoid valve V1 and embeds area 5.
Step 105：When the photoelectric sensor (b2) being arranged on enzyme mark primary antibody embedding 5 liquid outlet of area detects that external sample is dilute
When releasing liquid, solenoid valve V1 is closed, opens solenoid valve V5, and positive pressure and negative-pressure sucking are sequentially generated by plunger pump 19 so that blood
Clearly, external dilution liquid, the enzyme mark primary antibody embedded in advance are embedded in enzyme mark primary antibody and are flowed back and forth between area 5 and the first mixing pipeline 6
It redissolves, obtains the first mixed liquor.
Step 106：First mixed liquor is inhaled into magnetic mark secondary antibody embedding area 7, and makes first to mix by the second mixing pipeline 8
It closes liquid to be combined with antigen-antibody, the reactant of formation is captured by magnetic bead, and magnetic bead is by the magnet adsorption in 7 outside of magnetic mark secondary antibody embedding area
And stablize in magnetic mark secondary antibody embedding area 7, remaining reaction solution is under the negative-pressure sucking of plunger pump 19 through liquid driven power entrance
Chip is discharged, then carries out next cleaning step.
Step 107：Solenoid valve V5 is closed, and opens solenoid valve V3, exterior washings liquid is made to enter magnetic mark secondary antibody embedding area
7, and the note that the photoelectric sensor (c1, c2) being arranged on 7 inlet of area and liquid outlet controls cleaning solution is embedded by magnetic mark secondary antibody
Enter amount.
Step 108：After external cleaning solution and magnetic bead clean repeatedly, magnet 25a, 25b adsorb magnetic bead, are produced by plunger pump
Liquid suction after cleaning is discharged in external waste liquid pool 24 by raw negative-pressure sucking.
Step 109：Solenoid valve V3 is closed, solenoid valve V2 is opened, external substrate luminescent solution is made to enter the embedding of magnetic mark secondary antibody
Area 7, and pass through the injection rate of photoelectric sensor (c1, c2) control substrate luminescent solution.
Step 110：After substrate luminescent solution is fully reacted with the antigen-antibody on magnetic bead, reaction solution, reaction solution quilt are obtained
Chemiluminescence detection area 9 is transported, to complete chemiluminescence detection；Wherein, on 9 inlet of chemiluminescence detection area and liquid outlet
The photoelectric sensor (d1, d2) of setting is used to detect capacity and the position of reaction solution.
Reaction principle in the chemiluminescence micro-fluidic chip of the present embodiment between substance is the same as magnetic particle immunochemiluminescence
Reaction principle, i.e. antigen in sample by with enzyme mark primary antibody (primary antibody is marked with the catalytic groups such as HRP, AP) combine, then with
Magnetic mark secondary antibody (secondary antibody is fixed on magnetic bead) combines and forms double antibodies sandwich compound, and magnet adsorption magnetic bead washes unbonded
Antigen and enzyme mark primary antibody, substrate reactions liquid is added, the enzymes group catalysis substrate reactions liquid such as HRP, AP marked on primary antibody shines.
Luminous intensity and the amount of antigen are directly proportional.
Embodiment 3
Referring to FIG. 5, the present invention provides the liquid driving devices of function described in achievable embodiment 1 or 2.In this reality
It applies in example, liquid driving device is plunger pump 19.
For structure, liquid driving device may be configured as a variety of, such as existing syringe pump, diaphragm pump, peristaltic pump, all
It is that by and liquid is driven to the presumptive area to chip under pressure, protection scope of the present invention should all be fallen into.
Although syringe pump, diaphragm pump, peristaltic pump can drive liquid to flow, they cannot control liquid and stop in specific position well
It stays, and plunger pump can preferably solve the problems, such as this.Being suitable for the invention plunger pump can be ripe for those skilled in the art
The plunger pump known, generally includes pump chamber 194 and plunger 193, and pump chamber 194 is equipped with inlet 191 and liquid outlet 192, plunger
193 top is inserted into pump chamber, and plunger 193 is reciprocating in its axial direction along the inner wall of pump chamber 194；Inlet 191 goes out
Valve V4, V6 are respectively equipped at liquid mouth 192.Since plunger pump is applied to imbibition, drain, two be arranged on pump chamber by more
Mouth is commonly known as " inlet and liquid outlet ", but it should be recognized that " inlet and liquid outlet " herein is not limited to use
In feed liquor and going out liquid, in the present embodiment, when plunger pump work, after the valve V4 at inlet 191 is opened, plunger is transported downwards
Dynamic, liquid closes on the pressure of one end of plunger pump inlet 191 and becomes smaller at this time, causes liquid both ends to generate pressure difference, liquid exists
It is moved to 191 direction of inlet under the action of pressure difference, when liquid reaches pre-position, opens the valve at liquid outlet
V6 so that chip interior is connected to outside atmosphere, liquid both sides respectively both sides air (wherein the air of side through liquid outlet,
Inlet enters chip interior, and the air of the other side can (such as injection port or the air that is separately arranged be to logical from air inflow end mouth
Road) enter chip interior) under the action of, pressure keeps balance, liquid that can rest on pre-position.
Embodiment 4
It please refers to Fig.1 and Fig. 3, the present invention provides the liquid quantitative chambers of function described in achievable embodiment 1 or 2.
It should be noted that the liquid quantitative chamber of the present invention can realize that " liquid that need to be quantified is from liquid quantitative chamber
Inlet influent quantify chamber, reach liquid outlet after hydraulically full quantitative chamber ", shape and structure can be according to need
It is selected, the present invention imposes any restrictions not to this, such as it can be pipe shape, polygonal shape etc..
Realize that " liquid that need to be quantified flows into the liquid quantitative chamber from the inlet of the liquid quantitative chamber, is full of
Reach the liquid outlet after the liquid quantitative chamber " mode there are many, such as control liquid quantitative chamber width and height
Degree carries out hydrophilic and hydrophobic processing etc. on the surface of liquid quantitative chamber.
In the present embodiment, the liquid quantitative chamber is the chamber of hexagonal structure.Optionally, the liquid quantitative chamber
The inlet and liquid outlet of room are respectively that two of the hexagonal structure are diagonal；Described two diagonal angles are less than 120 °.
Optionally, the width of the inlet of liquid quantitative chamber is 0.3-3mm (preferably 0.8-1.5mm), is highly
0.3-3mm；The width of the liquid outlet of liquid quantitative chamber is 0.3-3mm (preferably 0.8-1.5mm), is highly 0.3-3mm.Into
Liquid mouth width spend wide or narrow, excessive height or it is too low be unfavorable for quantitative progress, when inlet width is wide or high spends
Gao Shi, be easy to cause liquid can not hydraulically full quantitative chamber flow to its liquid outlet, cannot achieve accurate liquid in this way and determine
Amount, and when inlet width is narrow or height is too low, then it needs accordingly to increase length to meet the requirement of volume, in this way may
It can lead to the increase of chip length and the increase of chip volume.
Optionally, the surface of liquid quantitative chamber is the surface formed after hydrophilic surface is modified；Liquid quantitative chamber
The width of the inlet of room is 0.3-5mm, is highly 0.3-3mm；The width of the liquid outlet of liquid quantitative chamber is 0.3-5mm,
Height is 0.3-3mm.Hydrophilic surface modification includes but not limited to plasma, hydroxylating, carboxylated modification.Liquid quantitative chamber
Surface carry out Hydrophilic modification after, be more advantageous to the filling of liquid in the cavity, at this time can larger fluid appropriate quantify chamber
The inlet of room, the width of liquid outlet, so as to reduce the length of liquid quantitative chamber and chip.
Optionally, the surface of liquid quantitative chamber is the surface formed after hydrophobic surface is modified, liquid quantitative chamber
The width of the inlet of room is 0.3-2mm, is highly 0.3-3mm；The width of the liquid outlet of liquid quantitative chamber is 0.3-2mm,
Height is 0.3-3mm.Hydrophobically modified includes but not limited to hydrophobicity physical modification, hydrophobic chemical modification (such as nano-particle
Coating, the alkyl etc. for lengthening chain).The surface of liquid quantitative chamber can prevent liquid wall built-up after hydrophobic surface is modified, and
It can guarantee and reach liquid outlet after liquid is full of in liquid quantitative chamber.
Embodiment 5
Fig. 2 is referred to, the present invention provides the Liquid identification site of function described in achievable embodiment 1 or 2 and liquid
Identification device.
It should be noted that liquid is known for positioning or fixing liquid identification device, the present invention in Liquid identification site
The structure of other device is not restricted, as long as the identification of liquid can be realized.Such as the patent Shen of Publication No. " 105214744A "
Please disclosed in liquid sensing device can be used as the present invention Liquid identification device, but such liquid sensing device structure compared with
For complexity, conductive pin needs to be built into chip interior, and conductive pin is contacted with reaction liquid, can influence reality in any case
Test as a result, and chip prepare difficulty it is larger, provide a kind of preferred Liquid identification device in the present embodiment.
In the present embodiment, Liquid identification site includes light source life for positioning Liquid identification device, Liquid identification device
At module 28 and photoelectric sensor 29；Liquid identification site includes for the upper site of positioned light source generation module 28 and for fixed
The lower site of position photoelectric sensor 29, upper site and lower site are respectively arranged on the outside of chip body, upper site, corresponding feed liquor
Mouth or liquid outlet, the perpendicular line in lower site are laid successively.Correspondingly, light source generation module 28, corresponding inlet or liquid outlet,
First photoelectric sensor, 29 perpendicular line is laid successively.Since Liquid identification device can be set to liquid quantitative chamber or detection chambers
Inlet or liquid outlet at, therefore " corresponding inlet or liquid outlet " herein correspond to liquid quantitative chamber or test chamber
The inlet or liquid outlet of room；For example, when Liquid identification device is arranged in the liquid outlet of liquid quantitative chamber, light source generates mould
Block, the liquid outlet of liquid quantitative chamber, the perpendicular line of photoelectric sensor are laid successively；When the inlet of liquid quantitative chamber is arranged
When Liquid identification device, light source generation module, the inlet of liquid quantitative chamber, the perpendicular line of photoelectric sensor are laid successively；
When Liquid identification device is arranged in the liquid outlet of sample amounts chamber, light source generation module, the liquid outlet of sample amounts chamber, light
The perpendicular line of electric inductor is laid successively.
Using optical sensing come to Liquid identification, quantitative and control, relative to the way of contact of conducting type, the method is reduced
Intervention of the metal to reaction system in chip can be improved detection efficiency, and then improve quantitative accuracy, while such liquid
Body identification device can be set to outside micro-fluidic chip, convenient for being fixed in instrument, without being arranged on chip, reduce chip
Difficulty of processing.In use, only light source generation module and photoelectric sensor alignment liquid recognition site need to be placed.Specifically
Ground, chip body include top plate 1 and bottom plate 20；Top plate 1 is connected with the stacking of bottom plate 20；Top plate 1 and the junction of bottom plate 20 are arranged
There are main fluid passageway and multiple function chambers；The setting of light source generation module 28 is located in and the inlet of liquid quantitative chamber or goes out
The surface of the corresponding position of the corresponding top plate of liquid mouth 1, photoelectric sensor 29 be located in the inlet of liquid quantitative chamber or
The underface of the corresponding position of the corresponding bottom plate of liquid outlet 20.
Light source generation module can provide the module of light source, can be LED, halogen lamp, laser lamp etc..In the photograph of light source
It penetrates down, since gas, liquid are to the transmissivity and refractive index difference of light, the light intensity for being irradiated to photoelectric sensor is different, photoelectricity
Inductor can identify gas and liquid, to distinguish whether liquid arrives the point of induction., when liquid flow to inlet or goes out
When liquid mouth, Liquid identification device can be identified quickly, to control liquid driving device.
Embodiment 6
The embodiments of the present invention also provide a kind of analytical instrument with micro-fluidic chip comprising apparatus frame, extremely
Micro-fluidic chip in few a reagent storage pool, liquid driving device, detection device and any of the above embodiment；Wherein, micro-
Fluidic chip is mounted in the apparatus frame；Liquid driving device is connected with the liquid driven power entrance of micro-fluidic chip；Examination
Agent storage pool can be connected to break-make with corresponding reagent inlet；Detection device is used to receive the detection that processing micro-fluidic chip is sent out
Signal.
Optionally, the liquid driving device is plunger pump；It is equipped on each reagent storage pool and outside air
The opening of connection.
The above is the preferred embodiment of the present invention, it is noted that for those skilled in the art
For, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also considered as
Protection scope of the present invention.
Claims (14)
1. a kind of micro-fluidic chip, which is characterized in that including chip body and the sample introduction being arranged in the chip body
Mouth, liquid driven power entrance, main fluid passageway and multiple function chambers；
The main fluid passageway is connected to the multiple function chamber, the injection port and the liquid driven power entrance respectively with institute
Main fluid passageway connection is stated, the liquid driven power entrance is for connecting liquid driving device to drive liquid to flow in or out work(
It can chamber；
At least one of the multiple function chamber is liquid quantitative chamber；The liquid quantitative chamber has scheduled appearance
Product, and is provided with Liquid identification site at the liquid outlet of liquid quantitative chamber, and the liquid that need to be quantified is from the liquid quantitative chamber
The inlet of room flows into the liquid quantitative chamber, full of reaching the liquid outlet after the liquid quantitative chamber.
2. micro-fluidic chip according to claim 1, which is characterized in that the liquid quantitative chamber includes reagent quantitative chamber
The inlet of room, the reagent quantitative chamber is connected to one end of reagent subchannel, the other end of the reagent subchannel and examination
Agent entrance is connected to.
3. micro-fluidic chip according to claim 1, which is characterized in that also set at the inlet of the liquid quantitative chamber
It is equipped with Liquid identification site.
4. micro-fluidic chip according to claim 1, which is characterized in that the liquid driving device is plunger pump.
5. micro-fluidic chip according to claim 1, which is characterized in that the function chamber includes detection chambers, described
Detection chambers have scheduled volume, and Liquid identification site is provided at the liquid outlet of the detection chambers, to be detected
Inlet of the liquid through the detection chambers flows into the detection chambers, full of reaching liquid outlet after the detection chambers.
6. micro-fluidic chip according to claim 5, which is characterized in that be also equipped at the inlet of the detection chambers
Liquid identification site.
7. according to micro-fluidic chip according to any one of claims 1 to 6, which is characterized in that use in the Liquid identification site
In positioning Liquid identification device；The Liquid identification device includes light source generation module and photoelectric sensor；
The Liquid identification site includes upper site for positioning the light source generation module and for positioning the light inductance
The lower site of device, the upper site and the lower site is answered to be respectively arranged on the outside of the chip body, the upper site is under
The position in site and corresponding liquid outlet or inlet are corresponding so that positioning after the light source generation module, go out accordingly
Liquid mouth or inlet, the perpendicular line of the photoelectric sensor are laid successively.
8. micro-fluidic chip according to claim 1, which is characterized in that the liquid quantitative chamber is hexagonal structure
Chamber.
9. micro-fluidic chip according to claim 1, which is characterized in that the width of the inlet of the liquid quantitative chamber
It is highly 0.3-3mm for 0.3-3mm；The width of the liquid outlet of the liquid quantitative chamber is 0.3-3mm, is highly 0.3-
3mm；Or
The surface of the liquid quantitative chamber is the surface formed after hydrophilic surface is modified；The liquid quantitative chamber
The width of inlet is 0.3-5mm, is highly 0.3-3mm；The width of the liquid outlet of the liquid quantitative chamber is 0.3-5mm,
Height is 0.3-3mm；Or
The surface of the liquid quantitative chamber is the surface formed after hydrophobic surface is modified, the liquid quantitative chamber
The width of inlet is 0.3-2mm, is highly 0.3-3mm；The width of the liquid outlet of the liquid quantitative chamber is 0.3-2mm,
Height is 0.3-3mm.
10. micro-fluidic chip according to claim 1, which is characterized in that the chip body includes top plate and bottom plate；Institute
It states the stacking of top plate and the bottom plate to connect, the junction of the top plate and the bottom plate is provided with the main fluid passageway and described
Multiple function chambers.
11. micro-fluidic chip according to claim 10, which is characterized in that the bottom plate is smooth tablet, the top
Micropore, microchannel or microcavity body are provided on plate to form the injection port, liquid driven power entrance, main fluid with the bottom plate
Channel or function chamber.
12. micro-fluidic chip according to claim 1, which is characterized in that the injection port and the liquid driven power enter
Mouth is separately positioned on the both ends of the main fluid passageway.
13. a kind of analytical instrument with micro-fluidic chip, which is characterized in that stored including apparatus frame, at least one reagent
Micro-fluidic chip described in any one of pond, liquid driving device, detection device and claim 1~12；Wherein, the miniflow
Chip is controlled to be mounted in the apparatus frame；The liquid driving device is connected with the liquid driven power entrance of micro-fluidic chip；
The reagent storage pool can be connected to break-make with corresponding reagent inlet；The detection device is for receiving processing micro-fluidic chip
The detection signal sent out.
14. the analytical instrument according to claim 13 with micro-fluidic chip, which is characterized in that the liquid driven dress
It is set to plunger pump；It is equipped with the opening being connected to outside air on each reagent storage pool.
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