CN207294793U - Micro-fluidic pcr detection system - Google Patents
Micro-fluidic pcr detection system Download PDFInfo
- Publication number
- CN207294793U CN207294793U CN201721166516.9U CN201721166516U CN207294793U CN 207294793 U CN207294793 U CN 207294793U CN 201721166516 U CN201721166516 U CN 201721166516U CN 207294793 U CN207294793 U CN 207294793U
- Authority
- CN
- China
- Prior art keywords
- micro
- sample
- fluidic pcr
- detection system
- diversion trench
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The utility model provides a kind of micro-fluidic PCR detection system, it includes micro-fluidic pcr chip, sample adding device, thermocirculator and fluorescent collecting device, wherein, the micro-fluidic pcr chip includes basal part, chamber portion, sealing cover and aerating mouth；The sample adding device is used to add detected sample at the hydrophilic unit, sealing oil reservoir is formed above the detected sample added, and inject gas into the reaction chamber from the aerating mouth；The thermocirculator is used to heat the substrate, so that the detected sample carries out thermal cycling amplification；The fluorescent collecting device is used to carry out the detected sample after thermal cycling amplification fluorescence signal acquisition, and can outwards export collection signal.Micro-fluidic PCR detection system described in the utility model can simplify PCR detection resolved ways, and operation is added easy to detected sample, can be beneficial to the progress of thermal cycling amplification reaction, and help to lift detection result.
Description
Technical field
Biomolecule detection technical field is the utility model is related to, more particularly to a kind of micro-fluidic PCR detection system.
Background technology
PCR (PCR) has become the new standard of molecular diagnosis field, and PCR method is from molecular level
Detection is realized by the design for the specific primer probe for being directed to gene order, compared with immunization method, PCR method can be big
The big accuracy and sensitivity for improving detection.Microflow control technique is brand-new point operated in micro-meter scale to fluid
Analysis technology, compared to conventional analysis method, it is with high throughput, automation, integrated and inexpensive etc. advantage.
At present, the combination of microflow control technique and round pcr is increasingly paid close attention to, and high pass is easily achieved by means of micro-fluidic
The advantage of screening is measured, round pcr can be applied to the fields such as Large-scale Screening and the diagnosis of molecular marked compound.Existing molecule is examined
The High Throughput Screening Assay of disconnected aspect mainly includes molecule hybridization (DNA chip technology), multiple PCR technique and liquid-phase chip technology
Deng.Molecular hybridization needs in advance to expand target gene, and then amplified production is hybridized with complementary probe sequences
Just it can detect afterwards as a result, it will also be rinsed and warm bath in addition, it is complex for operation step.Multiple PCR technique is needed to different
Target gene carries out fluorescence probe coding, not only needs high-resolution optical system, while more target simultaneous reactions, primer and
Probe design requirement is high, and be easy to cause cross reaction.Liquid-phase chip technology not only needs the fluorescence-encoded micro-beads system of complexity
Standby and covalent coupling technology, while it is also required to the operating procedure of complexity, its expensive equipment so that ordinary user is difficult to receive.
Utility model content
In view of this, the utility model is directed to a kind of micro-fluidic PCR detection system, with high-throughout available for carrying out
PCR detections use.
To reach above-mentioned purpose, what the technical solution of the utility model was realized in：
A kind of micro-fluidic PCR detection system, it includes：
Micro-fluidic pcr chip, the micro-fluidic pcr chip include basal part, chamber portion, sealing cover and aerating mouth, and institute
Stating basal part includes the substrate with heat transmitting, is laid in the hydrophobic layer of the base top, and with micro-array cloth
The multiple hydrophilic units being placed on the hydrophobic layer；The chamber portion is connected with the substrate, is configured to enclosing positioned at each described
Reaction chamber above hydrophilic unit；The sealing cover has translucency and is connected to the top in the chamber portion, with composition pair
The sealing of the reaction chamber；The aerating mouth is located on the sealing cover or chamber portion one, to calm the anger with outside
Source connects, and in injecting gas in the reaction chamber；
Sample adding device, the sample adding device are used to add detected sample at the hydrophilic unit, are treated in what is added
Detect and sealing oil reservoir is formed above sample, and gas is injected into the reaction chamber from the aerating mouth；
Thermocirculator, the thermocirculator be used for the substrate is heated so that the detected sample into
Row thermal cycling amplification；
Fluorescent collecting device, the fluorescent collecting device are used to carry out the detected sample after thermal cycling amplification glimmering
Light signal collection, and collection signal can outwards be exported.
Further, the reagent for including primer having in dry shape is deposited at least at one in each hydrophilic unit.
Further, the reagent is formed by ink-jet printer point sample at the hydrophilic unit, and through drying process
Dry shape.
Further, the drying process mode includes vacuum freeze drying or room temperature spontaneously dries.
Further, the temperature of the vacuum freeze drying is -85~-75 DEG C, and vacuum is 0.008~0.012Mpa.
Further, the substrate is hydrophilic material, the hydrophilic unit as described in hollow out hydrophobic layer and be exposed to institute
The substrate stated outside hydrophobic layer is formed.
Further, the material of the substrate includes glass or silicon chip.
Further, the material of the hydrophobic layer includes silicon fluoride or hmds.
Further, micro- trap battle array that the surface of the substrate at each hydrophilic unit is formed for smooth flat or etching
Row.
Further, the etching mode includes wet etching or dry etching.
Further, the basal part is prepared by following step：
Step s1, substrate is provided；
Step s2, in forming hydrophobic layer in the substrate；
Step s3, in forming photoresist layer on the hydrophobic layer, and formed by photolithographicallpatterned on the photoresist layer
In the pattern of multiple Openworks shapes of microarray arrangement；
Step s4, bombarded by plasma machine, remove the hydrophobic layer of each area of the pattern, make the area of the pattern
Substrate forms the hydrophilic unit outside；
Step s5, the photoresist layer is peeled off.
Further, further included before the step s5：
Step s50, etch to form micro- trap array in the substrate surface of the hydrophilic unit.
Further, it is that the substrate and the hydrophobic layer material are together put into heater box in step s2, constant temperature is kept,
And generate the hydrophobic layer in substrate surface reaction.
Further, specifically included in the step s3：
Step s31, in spin coating photoresist on the hydrophobic layer, and dry and form photoresist layer；
Step s32, in litho machine, UV exposure-processeds are carried out to the photoresist layer under mask plate；
Step s33, it is put into developer solution and carries out development treatment, the pattern is formed in photoresist layer；
Step s34, baking and curing.
Further, the chamber portion is made of translucent material.
Further, the sealing cover and the material in the chamber portion include PP, PET or PC.
Further, the substrate is connected with the chamber portion through adhesive is Nian Jie.
Further, the adhesive includes epoxide-resin glue or uv-curable glue.
Further, the sample adding device includes：
Fixed frame, the fixed frame are used to load the micro-fluidic pcr chip；
Brake bar, is slideably positioned on the fixed frame；
Sample diversion trench, is arranged on the brake bar, and the sample diversion trench has an open-topped liquid storage cylinder, and in
The bottom of the sample diversion trench is equipped with the outlet in slit-shaped with liquid storage cylinder perforation；
Seal Oil diversion trench, is arranged on the brake bar, and has the structure identical with the sample diversion trench；
Driving mechanism, is arranged between the fixed frame and the brake bar, to drive described in the brake bar drive
Sample diversion trench and the Seal Oil return guide trough are into the slip relative to the micro-fluidic pcr chip；
Control mechanism, the control mechanism are connected with the driving mechanism, to control the driving mechanism to act；
Aerating interface, the aerating interface is multiple to be connected respectively with external air source, and each aerating interface difference
Open top with the aerating mouth and the sample diversion trench and Seal Oil diversion trench communicates, with to the reaction chamber and institute
State injection gas in sample diversion trench and Seal Oil diversion trench.
Further, the driving mechanism includes guide rail, and be sequentially connected with the brake bar, drive the braking
The stepper motor that bar is slided along the guide rail.
Further, inject the indoor gas pressure of the reaction chamber and be more than 0.1MPa.
Further, it is inert gas or air to inject the indoor gas of the reaction chamber.
Further, the aerating interface being connected in the open top with the sample diversion trench and Seal Oil diversion trench
Place is equipped with pressure-regulating valve part.
Further, the thermocirculator includes：
Supporting rack, for carrying the micro-fluidic pcr chip；
Thin film heater, abuts on support frame as described above, and with the substrate of the micro-fluidic pcr chip；
Power controling machine structure, is electrically connected with external power supply and the film heating piece, to control to the film heating piece
Power supply.
Further, the fluorescent collecting device includes：
Light source；
First optical filter, on the outgoing light direction of the light source, and filters the emergent light of the light source；
Dichroscope, on the outgoing light direction of the light source, to form the light source emergent light to described micro-fluidic
The detected sample in the irradiation of pcr chip, and the micro-fluidic pcr chip fluoresces outside outgoing；
Image acquisition units, in the exit direction of the fluorescence, to be acquired to the fluorescence, and will can gather
Signal outwards exports；
Second optical filter, in the exit direction of the fluorescence, and is placed in the front side of described image collecting unit, with right
The fluorescence of outgoing is filtered.
Further, the light source is laser light source, and described image collecting unit is cmos sensor.
Relative to the prior art, the micro-fluidic PCR detection system of the utility model has the advantage that：
In the micro-fluidic PCR detection system of the utility model, by hydrophobic layer in micro-fluidic pcr chip being in microarray
Hydrophilic unit arrangement, can realize that high flux screening detects, and can be fixed by different indexs (specific primer and probe)
In the specific response location of substrate, so as to be differentiated by space, it is not required to carry out spectrally resolved, the energy of multicolor fluorescence coding
It is enough to simplify detection resolved way.
Meanwhile by the setting of hydrophobic layer and hydrophilic unit in the utility model, when sample adds, by hydrophobic layer and
The hydrophobe effect of hydrophilic unit, can automatically form independent sample drop, so as to easy to be checked at each hydrophilic unit
Sample adds operation.
In addition, by the sealing of reaction chamber and the setting of aerating mouth in the utility model, when detection, can pass through injection
Gas and is kept high pressure conditions, can be heated with this in PCR anti-so that reaction chamber gas pressure inside is more than the saturated vapour pressure of water
Suppress the evaporation of moisture during answering, can also significantly reduce the volume of each sample drop.Thus, it may be such that the reagent that detection uses
Consumption reduce, more PCR reaction members can be also integrated on unit area, and the number of PCR Testing index can be improved
Amount and degree of parallelism so that detection device integrated level is high, to contribute to the reduction of testing cost.
And the dense of sample to be tested is also remarkably improved by reducing the volume of single sample drop in the utility model
Degree, can be conducive to the progress of PCR thermal cycling amplifications reaction, when can improve the sensitivity of pcr chip, and can greatly shorten reaction
Between, with i.e. detectable positive signal within a short period of time, lift detection efficiency.
In conclusion the micro-fluidic PCR detection system of the utility model passes through the application of micro-fluidic pcr chip, Yi Jitong
The setting of sample adding device, thermocirculator and fluorescent collecting device is crossed, can realize that high flux screening detects, PCR inspections can be simplified
Resolved way is surveyed, operation is added easy to detected sample, the progress of thermal cycling amplification reaction can be beneficial to, contribute to lifting to examine
Effect is surveyed, and detection reagent consumption is few, device integration is high, testing cost is low, and have good practicality.
Brief description of the drawings
The attached drawing for forming the part of the utility model is used to provide a further understanding of the present invention, this practicality is new
The schematic description and description of type is used to explain the utility model, does not form the improper restriction to the utility model.
In attached drawing：
Fig. 1 is the structure diagram of the micro-fluidic pcr chip described in the utility model embodiment；
Fig. 2 is the preparation flow figure of the basal part described in the utility model embodiment；
Fig. 3 is the structure diagram of the sample adding device described in the utility model embodiment；
Fig. 4 is the structure diagram of the sample diversion trench described in the utility model embodiment；
Fig. 5 is that the micro-fluidic pcr chip described in the utility model embodiment adds the knot after detected sample and Seal Oil
Structure schematic diagram；
Fig. 6 is the operating diagram of the micro-fluidic PCR detection system described in the utility model embodiment；
Description of reference numerals：
1- substrates, 2- chambers portion, 3- sealing covers, 4- aerating mouths, 5- hydrophobic layers, 6- hydrophilic units, 7- reaction chambers, 8- glue
Glutinous agent, 9- photoresist layers, 10- mask plates, 11- sample diversion trenches, 12- Seal Oil diversion trenches, 13- brake bars, 14- fixed frames
Frame, 15- guide rails, 16- stepper motors, 17- sample drops, 18- sealing oil reservoirs, 19- supporting racks, 20- thin film heaters, 21- light
Source, 22- image acquisition units, the first optical filters of 23-, 24- dichroscopes, the second optical filters of 25-, 100- chips, 111- liquid storages
Chamber, 112- open tops, 113- outlets.
Embodiment
It should be noted that in the case where there is no conflict, the feature in embodiment and embodiment in the utility model can
To be mutually combined.
The utility model will be described in detail below with reference to the accompanying drawings and embodiments.
The present embodiment is related to a kind of micro-fluidic PCR detection system, which has generally comprised micro-fluidic PCR cores
Piece, further includes for adding detected sample into the micro-fluidic pcr chip, is formed above the detected sample added close
Oil sealing layer, and into micro-fluidic pcr chip inject gas sample adding device, for being heated to micro-fluidic pcr chip, with
Detected sample is set to carry out the thermocirculator of thermal cycling amplification, and for being carried out to the detected sample after thermal cycling amplification
Fluorescence signal acquisition, and the fluorescent collecting device that the signal of collection can be outwards exported.
Include basal part, chamber portion and sealing cover and aerating mouth in the micro-fluidic pcr chip structure of the present embodiment, and as schemed
Shown in 1, wherein, basal part includes the substrate 1 with heat transmitting, and tiling is arranged at the hydrophobic layer 5 at the top of substrate 1, with
And multiple hydrophilic units 6 on hydrophobic layer 5 are arranged in micro-array.Chamber portion 2 is connected with substrate 1 and encloses and be configured to position
Reaction chamber 7 above each hydrophilic unit 6, sealing cover 7 are connected to the top in chamber portion 2, to form to the reaction chamber 7
Sealing, and sealing cover 7 is translucency.Aerating mouth 4 is then located on sealing cover 3, and it is used for having pressurized air source to connect with outside, and
Gas is injected into reaction chamber 7.
Particularly, substrate 1 is made of hydrophilic material in the present embodiment, it is preferred that the material for forming substrate 1 can be parent
Water-based glass or silicon chip, and the material that hydrophobic layer 5 is formed in the present embodiment can be then silicon fluoride or hmds.This
Outside, can be by the hollow out hydrophobic layer 5, so that the hollow out position by hydrophobic layer 5 in the present embodiment as a kind of preferable structure type
Put the hydrophilic of the microarray arrangement for having hydrophilic substrate 1 composition on hydrophobic layer 5 being exposed to outside hydrophobic layer 5 at place
Unit 6, at this time each hydrophilic unit 6 be rendered as a bowl configurations.
In the present embodiment for the hydrophilic unit 6 directly constituted by substrate 1, further, in a particular application may be used
So that the surface of the substrate 1 at each hydrophilic unit 6 is smooth flat, and except for smooth flat, also can by etching processing and
So that forming micro- trap array exposed to the surface of outer substrate 1, etching processing at this time uses existing wet etching or dry method
Etch tool.
For the basal part of constituted above when actually preparing, it can be as shown in Figure 2, at this time with substrate 1 in the present embodiment
Using silicon chip, exemplified by 5 material of hydrophobic layer uses silicon fluoride, choose the silicon chip being ready to as substrate 1 first, then by silicon chip and
Silicon fluoride is together put into heater box, and a hour is kept at a temperature of 80 DEG C so that silicon fluoride become steam and with silicon chip table
React to form hydrophobic layer 5 in face.
After hydrophobic layer 5 is formed, then, the spin coating photoresist on hydrophobic layer 5, and dry at 110 DEG C one minute from
And photoresist layer 9 is formed, then the silicon chip formed with photoresist layer 9 is put into litho machine, to photoresist under mask plate 10
Layer 9 carries out UV exposure-processeds, just places into silicon chip after exposure-processed and development treatment is carried out in developer solution, at this time can be in photoetching
Form the pattern of multiple Openworks shapes in microarray arrangement on glue-line 9, and the pattern concretely circular, rectangle or other shapes
Shape.After the pattern of microarray arrangement is formed, then silicon chip is positioned under plasma machine and is bombarded, remove each area of the pattern
The hydrophobic layer 5 that silicon fluoride outside photoresist layer 9 is formed so that the silicon chip of each area of the pattern is externally exposed, so just
Form the hydrophilic unit 6 being made of substrate 1.
After the silicon chip exposure of each area of the pattern, what the present embodiment still can be shown in Fig. 2 can also use wet etching or dry
The mode of method etching etches to form micro- trap array in the surface ion exposed to outer silicon chip.Certainly, if desired so that hydrophilic list
The surface of substrate 1 at member 6 is smooth flat, also the step for removable ion etching, is just carried out after plasma bombardment
The stripping of photoresist layer 9.
And after each hydrophilic unit 6 is formed, then, it is also necessary to be rinsed silicon chip by acetone, to peel off residue
Photoresist layer 9, then after the removal of photoresist layer 9, recycle isopropyl acetone and deionized water to be cleaned successively to silicon chip, and
Silicon chip is dried at 100 DEG C.
In the present embodiment, the chamber portion 2 being connected with substrate 1 also by being made of the material of translucency, and chamber portion 2 with it is same
Sealing cover 3 with translucency can use such as PP, PET or PC polymer plastic to be processed through Shooting Technique and be made.And in base
In connection mode between bottom 1 and chamber portion 2, both can be bonded together by adhesive 8 in the present embodiment, and the adhesive
8 can specifically use epoxide-resin glue or uv-curable glue.
In the present embodiment, sealing cover 3 can be connected to the top in chamber portion 2 by way of clamping, be arranged on sealing cover 3
Aerating mouth 4 can specifically use female Luer or other existing air-path interface structures, can be connected by tracheae with external air source.
In addition, except aerating mouth 4 is arranged on sealing cover 3, certainly on the premise of detection is not influenced, aerating mouth 4 can also be arranged
Sidepiece or bottom in chamber portion 2.
In the micro-fluidic pcr chip of the present embodiment, further, at it in use, in each hydrophilic unit 6 formed
In, also deposited at least one hydrophilic unit 6 formed with the reagent in dry shape for including primer.The reagent is detecting
When, generally to be made of primer and probe, and be at hydrophilic unit 6 arrange when, reagent can pass through ink-jet printer point sample
1 surface of substrate at hydrophilic unit 6, then according to the type of reagent, then it is naturally dry by vacuum freeze drying or room temperature
Dry mode so that reagent drying.Wherein, for vacuum freeze drying mode, its operating condition in the present embodiment
It is temperature at -85~-75 DEG C, may be, for example, -80 DEG C, vacuum is 0.008~0.012Mpa, may be, for example, 0.01MPa.
After deposition includes the reagent of primer and probe, the micro-fluidic pcr chip of the present embodiment can carry out test sample to be checked
Product add, and thermal cycling amplification, and carry out PCR detection operations.
The sample adding device of the present embodiment is specifically used for adding test sample to be checked at the hydrophilic unit 6 in micro-fluidic pcr chip
Product, and sealing oil reservoir is formed above the detected sample added, and gas is injected into reaction chamber 7 from aerating mouth
Sample adding device.And the thermocirculator of the present embodiment is then specifically used for heating the substrate 1 in micro-fluidic pcr chip, with
Detected sample is set to carry out thermal cycling amplification.
Specifically, a kind of example arrangement of the present embodiment sample adding device is illustrated in figure 3, it includes fixed frame
14, fixed frame 14 is a platform structure, for loading the micro-fluidic pcr chip that need to be carried out detected sample and add (referred to as
Chip), chip 100 can be by the groove structure that is formed on fixed frame 14, and is positioned on fixed frame 14.In fixed frame
The brake bar 13 for sliding arrangement is provided with frame 14, is then provided with brake bar 13 and is respectively used to the sample for adding detected sample
Diversion trench 11 and for adding the Seal Oil diversion trench 12 of Seal Oil, and then also set up between brake bar 13 and fixed frame 14
The driving mechanism for being useful for driving brake bar 13 to drive two diversion trenches to be slided relative to chip 100, the driving mechanism is also with one
Control mechanism is connected, to be controlled by action of the control mechanism to driving mechanism.
Above-mentioned driving mechanism specifically includes two be fixed on fixed frame 14 and is arranged side by side on core in the present embodiment
The guide rail 15 of 100 two opposite sides of piece, and another guide rail 15 slided on two guide rails 15 is driven by stepper motor 16,
The guide rail 15 is arranged vertically with two guide rails 15 below, and brake bar 13 is i.e. on the guide rail 15, and brake bar 13 also leads to
Driving for 1 one stepper motors 16 is crossed, and can realize the slip of brake bar 13 and two diversion trenches along this guide rail 15, stepping electricity
The form slided along guide rail 15 under machine 16 drives refers to existing structure.Thus, each guide rail 15 and stepper motor 16 are passed through
Set, just can realize the movement in any position of two diversion trenches relative to chip 100 in the two-dimensional direction.
As shown in Figure 4, it has a hollow liquid storage to a kind of example arrangement of the sample diversion trench 11 of the present embodiment
Chamber 111, the top of sample diversion trench 11 is provided with the open top 112 connected with liquid storage cylinder 111, at the bottom of sample diversion trench 11
Portion, which is then provided with, to connect with liquid storage cylinder 111 and in the outlet 113 of elongated slit-shaped.Pass through the outlet 113 of the slit-shaped
Set, may be such that the detected sample in injection liquid storage cylinder 111 is formed at outlet 113 and suspended by means of surface tension effects
Hanging drop in 11 bottom of sample diversion trench.
At this time, the fixed position by adjusting sample diversion trench 11 on brake bar 13, and to be located at sample diversion trench
The hanging drop of 11 bottoms is contacted with 1 surface of substrate in chip 100, and is driven by stepper motor 16, and causes sample water conservancy diversion
During groove 11 is slided along chip 100, with sample diversion trench 11 slow mobile, (speed is in 0.01mm/s~100mm/s
Between), due to attracting caused by the hydrophily of the substrate 1 at the hydrophobicity and hydrophilic unit 6 of hydrophobic layer 5 detected sample
Difference in power, detected sample will be split off open and be fixed on at hydrophilic hydrophilic unit 6, thus can be in chip
Independent sample drop is formed on 100.
The structure of Seal Oil diversion trench 12 is identical with sample diversion trench 11 in the present embodiment, and its adding in Seal Oil
Also it is identical to add process with above-mentioned detected sample.But, it should be noted that in order to ensure detected sample and
The continuation of Seal Oil adds, in the present embodiment during diversion trench is moved with brake bar, it is also desirable to passes through open top
112 inject the gas of certain pressure into liquid storage cylinder 111, with this detected sample or Seal Oil are smoothly flowed out,
And form hanging drop in diversion trench bottom.
The control mechanism being connected with driving mechanism of the present embodiment can specifically use existing microcontroller or PLC, Yi Jiwei
Put, the electric controling part such as displacement sensor.And the sample adding device of the present embodiment then further included it is multiple respectively with outside
The aerating interface that source of the gas is connected, the above-mentioned gas injection into liquid storage cylinder 111 are real by the connection of the aerating interface and open top 12
It is existing, meanwhile, which is also used for connecting with the aerating mouth 4 on chip 100, to be injected in the reaction chamber 7 to chip 100
Gas.
Wherein, which is specifically made of the existing gas path joint being connected with gas piping, and the present embodiment
In gas from aerating interface to reaction chamber 7 that injected by can be inert gas or air, in its reaction chamber 7 after injecting
Gas pressure should be not less than 0.1MPa.In addition, in order to the gas in injection sample diversion trench 11 and Seal Oil diversion trench 12
Control is adjusted in pressure so that detected sample and Seal Oil realize smoothly outflow, with sample diversion trench 11 or close
The connected each aerating interface of oil sealing diversion trench 12 is also equipped with pressure-regulating valve part, which can use existing gas
Road valve body structure, and be preferably the automatically controlled valve body with pressure transmitter, to reach closed loop regulating effect.
The chip 100 of the present embodiment adds the state after detected sample and Seal Oil by sample adding device can be as in Fig. 5
It is shown, the independent sample drop 17 being made of detected sample is formd at each hydrophilic unit 6 at this time, Seal Oil is covered in
On hydrophobic layer 5 and each sample drop 17, the sealing oil reservoir 18 that can obstruct moisture evaporation is formd.And on sealing oil reservoir 18
In reaction chamber 7, after injecting gas of the pressure not less than 0.1MPa by aerating mouth 4, sample drop can be also further prevented
Moisture in 17 punctures sealing oil reservoir 18 in thermocirculator heating process and volatilizees.
In the present embodiment as shown in Figure 6, thermocirculator specifically includes the supporting rack 19 for carrying chip 100,
And the thin film heater 20 on supporting rack 19 is arranged at, supporting rack 19 is also a platform structure, and chip 100 is being positioned over support
Substrate 1 is abutted against with thin film heater 20 after on frame 19, and thin film heater 20 is then electrically connected with power controling machine structure, the power supply
Control mechanism is also electrically connected with external power supply at the same time, it is heated substrate 1 can power to thin film heater, and is caused
Detected sample in sample drop 17 carries out thermal cycling amplification reaction.In addition, the power controling machine structure of the present embodiment can also lead to
Cross control of the set temperature transmitter realization to heating-up temperature.
Still as shown in Figure 6, the fluorescent collecting device of the present embodiment includes light source 21, and positioned at 21 emergent light of light source
The first optical filter 23 and dichroscope 24 on direction, the light that the first optical filter 23 sends light source 21 filter, dichroic
Mirror 24 then reflects the emergent light of light source 21, so that the emergent light of light source 21 is irradiated on chip 100, and makes sample drop
Fluorescence is produced at 17.And the fluorescent collecting device of the present embodiment further includes the fluorescence exit direction sent positioned at sample drop 17
On the first optical filter 25 and image acquisition units 22.
The second optical filter 25 is used to filter the fluorescence of outgoing in the present embodiment, and image acquisition units 22 can specifically adopt
With existing cmos sensor, and as a preferred embodiment, light source 21 can then use laser light source.
The micro-fluidic PCR detection system of the present embodiment applies example when being detected, as one of those, makes first
The standby chip 100 as in embodiment one, chip 100 can carry out the deposition of reagent at random at each hydrophilic unit 6 in preparing, then
In injection detected sample in sample diversion trench 11, in injection Seal Oil in Seal Oil diversion trench 12, then control pressure regulating valve
Part causes each diversion trench bottom to produce hanging drop, and causes hanging drop and the contact of 1 surface of substrate.Then, in the driving of stepper motor 16
Under, make brake bar 13 drive two diversion trenches slowly to move, sample diversion trench 11 forms the independent sample liquid at hydrophilic unit 6
Drop 17, Seal Oil diversion trench 12 then subsequently form the sealing oil reservoir 18 of covering arrangement.
Then, sealing cover 3 is connected, and inert gas or sky are injected into the reaction chamber 7 of chip 100 by aerating mouth 4
Chip 100, can be positioned on supporting rack 19 by gas, carry out thermal cycling amplification reaction to be heated by thin film heater 20, instead
Fluorescent collecting device can be passed through after the completion of answering and carry out fluorescent collecting.At this time, can find depositing by the fluoroscopic image of collection
There is fluorescence intensity at the hydrophilic unit 6 of reagent to strengthen, form the reaction site of the positive, the hydrophilic unit without depositing reagent
It is then negative positions at 6.
The above is only the preferred embodiment of the utility model only, is not intended to limit the utility model, all at this
Within the spirit and principle of utility model, any modification, equivalent replacement, improvement and so on, should be included in the utility model
Protection domain within.
Claims (12)
- A kind of 1. micro-fluidic PCR detection system, it is characterised in that including：Micro-fluidic pcr chip, the micro-fluidic pcr chip include basal part, chamber portion, sealing cover and aerating mouth, and the base Bottom includes the substrate with heat transmitting, is laid in the hydrophobic layer of the base top, and be arranged in micro-array Multiple hydrophilic units on the hydrophobic layer；The chamber portion is connected with the substrate, is configured to enclosing positioned at each described hydrophilic Reaction chamber above unit；The sealing cover has translucency and is connected to the top in the chamber portion, to form to described The sealing of reaction chamber；The aerating mouth is located on the sealing cover or chamber portion one, to have pressurized air source company with outside It is logical, and in injecting gas in the reaction chamber；Sample adding device, the sample adding device is used to add detected sample at the hydrophilic unit, to be detected in what is added Sealing oil reservoir is formed above sample, and gas is injected into the reaction chamber from the aerating mouth；Thermocirculator, the thermocirculator is used to heat the substrate, so that the detected sample carries out heat Cyclic amplification；Fluorescent collecting device, the fluorescent collecting device are used to carry out fluorescence letter to the detected sample after thermal cycling amplification Number collection, and can will collection signal outwards export.
- 2. micro-fluidic PCR detection system according to claim 1, it is characterised in that：In each hydrophilic unit at least its Deposition has the reagent for including primer in dry shape at one.
- 3. micro-fluidic PCR detection system according to claim 1, it is characterised in that：The chamber portion is by translucent material It is made.
- 4. micro-fluidic PCR detection system according to any one of claim 1 to 3, it is characterised in that：The substrate is parent Water-based material, the hydrophilic unit described in hollow out hydrophobic layer and the substrate outside the hydrophobic layer is formed.
- 5. micro-fluidic PCR detection system according to claim 4, it is characterised in that：It is described at each hydrophilic unit Micro- trap array that the surface of substrate is formed for smooth flat or etching.
- 6. micro-fluidic PCR detection system according to claim 1, it is characterised in that：The sample adding device includes：Fixed frame, the fixed frame are used to load the micro-fluidic pcr chip；Brake bar, is slideably positioned on the fixed frame；Sample diversion trench, is arranged on the brake bar, and the sample diversion trench has open-topped liquid storage cylinder, and in described The bottom of sample diversion trench is equipped with the outlet in slit-shaped with liquid storage cylinder perforation；Seal Oil diversion trench, is arranged on the brake bar, and has the structure identical with the sample diversion trench；Driving mechanism, is arranged between the fixed frame and the brake bar, to drive the brake bar to drive the sample Diversion trench and the Seal Oil return guide trough are into the slip relative to the micro-fluidic pcr chip；Control mechanism, the control mechanism are connected with the driving mechanism, to control the driving mechanism to act；Aerating interface, the aerating interface for be connected respectively with external air source it is multiple, and each aerating interface respectively with institute The open top for stating aerating mouth and the sample diversion trench and Seal Oil diversion trench communicates, with to the reaction chamber and the sample Injection gas in product diversion trench and Seal Oil diversion trench.
- 7. micro-fluidic PCR detection system according to claim 6, it is characterised in that：The driving mechanism includes guide rail, with And be sequentially connected with the brake bar, the stepper motor that drives the brake bar to be slided along the guide rail.
- 8. micro-fluidic PCR detection system according to claim 6, it is characterised in that：Inject the indoor gas of the reaction chamber Body pressure is more than 0.1MPa.
- 9. micro-fluidic PCR detection system according to claim 8, it is characterised in that：Inject the indoor gas of the reaction chamber Body is inert gas or air.
- 10. micro-fluidic PCR detection system according to claim 6, it is characterised in that：With the sample diversion trench and close The aerating interface that the open top of oil sealing diversion trench is connected is equipped with pressure-regulating valve part.
- 11. micro-fluidic PCR detection system according to claim 1, it is characterised in that：The thermocirculator includes：Supporting rack, for carrying the micro-fluidic pcr chip；Thin film heater, abuts on support frame as described above, and with the substrate of the micro-fluidic pcr chip；Power controling machine structure, is electrically connected with external power supply and the film heating piece, to control the confession to the film heating piece Electricity.
- 12. micro-fluidic PCR detection system according to claim 1, it is characterised in that：The fluorescent collecting device includes：Light source；First optical filter, on the outgoing light direction of the light source, and filters the emergent light of the light source；Dichroscope, on the outgoing light direction of the light source, to form the light source emergent light to the micro-fluidic PCR cores The detected sample in the irradiation of piece, and the micro-fluidic pcr chip fluoresces outside outgoing；Image acquisition units, in the exit direction of the fluorescence, to be acquired to the fluorescence, and can will gather signal Outwards output；Second optical filter, in the exit direction of the fluorescence, and is placed in the front side of described image collecting unit, with to outgoing The fluorescence filtered.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721166516.9U CN207294793U (en) | 2017-09-12 | 2017-09-12 | Micro-fluidic pcr detection system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721166516.9U CN207294793U (en) | 2017-09-12 | 2017-09-12 | Micro-fluidic pcr detection system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN207294793U true CN207294793U (en) | 2018-05-01 |
Family
ID=62447191
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201721166516.9U Active CN207294793U (en) | 2017-09-12 | 2017-09-12 | Micro-fluidic pcr detection system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN207294793U (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107603874A (en) * | 2017-09-12 | 2018-01-19 | 深圳市尚维高科有限公司 | Micro-fluidic pcr detection system |
| CN110195016A (en) * | 2019-07-17 | 2019-09-03 | 中国人民解放军军事科学院军事医学研究院 | Temperature control objective table and detection device |
| WO2021114042A1 (en) * | 2019-12-09 | 2021-06-17 | 彩科(苏州)生物科技有限公司 | Microsphere and microwell plate-based detection device and use method therefor |
-
2017
- 2017-09-12 CN CN201721166516.9U patent/CN207294793U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107603874A (en) * | 2017-09-12 | 2018-01-19 | 深圳市尚维高科有限公司 | Micro-fluidic pcr detection system |
| CN107603874B (en) * | 2017-09-12 | 2024-04-05 | 深圳市尚维高科有限公司 | Microfluidic PCR detection system |
| CN110195016A (en) * | 2019-07-17 | 2019-09-03 | 中国人民解放军军事科学院军事医学研究院 | Temperature control objective table and detection device |
| WO2021114042A1 (en) * | 2019-12-09 | 2021-06-17 | 彩科(苏州)生物科技有限公司 | Microsphere and microwell plate-based detection device and use method therefor |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107603874A (en) | Micro-fluidic pcr detection system | |
| CN107475074A (en) | Micro-fluidic pcr chip | |
| CN207294793U (en) | Micro-fluidic pcr detection system | |
| KR102426954B1 (en) | Convection PCR Amplification Detection System and Convection PCR Amplification Detection Method | |
| AU2019264667B2 (en) | Method of removing floatation liquid | |
| US11236233B2 (en) | Sol-gel based matrix | |
| US8418929B2 (en) | Temperature controlling apparatus and temperature controlling method | |
| GB2518103B (en) | Pre-processing/electrophoresis integrated cartridge, device and method | |
| CN108431199A (en) | Reaction treating device, reaction treatment container, reaction treatment method | |
| CN207276595U (en) | Micro-fluidic pcr chip | |
| JP2009526969A (en) | Microfluidic devices for molecular diagnostic applications | |
| US20140295441A1 (en) | Cartridge interface module | |
| JP2009533048A (en) | Antifoam thin film for once-through cells | |
| Bog et al. | Large-scale parallel surface functionalization of goblet-type whispering gallery mode microcavity arrays for biosensing applications | |
| EP1643249A1 (en) | Bio-chip | |
| CN102886280A (en) | Microfluidic chip and application thereof | |
| CN103959052B (en) | The method for detecting specificity of analyte | |
| CN106198659A (en) | A kind of method depositing nanometer gold in micro-fluidic duct | |
| CN115093961A (en) | Multi-volume liquid drop digital LAMP nucleic acid absolute quantitative detection device and method and application | |
| CN109868220B (en) | Microfluidic dissolved oxygen detection chip and application thereof | |
| CN108579829B (en) | Pump-free micro-fluidic chip, preparation method thereof and portable biochemical analysis device | |
| CN112378972B (en) | Non-mark biosensor and manufacturing method thereof | |
| CN112063510A (en) | Structure of high-flux cell culture chip and manufacturing and using method thereof | |
| KR102365010B1 (en) | Multiplex pcr chip and multiplex-pcr detection method using the same | |
| JP4112285B2 (en) | Inspection method for biological substances |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |