CN106053467B - A kind of device and method for observing microlayer model - Google Patents
A kind of device and method for observing microlayer model Download PDFInfo
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- CN106053467B CN106053467B CN201610403838.4A CN201610403838A CN106053467B CN 106053467 B CN106053467 B CN 106053467B CN 201610403838 A CN201610403838 A CN 201610403838A CN 106053467 B CN106053467 B CN 106053467B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
- G01B11/00—Measuring arrangements characterised by the use of optical techniques
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
- G01B11/00—Measuring arrangements characterised by the use of optical techniques
- G01B11/24—Measuring arrangements characterised by the use of optical techniques for measuring contours or curvatures
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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- H—ELECTRICITY
- H04—ELECTRIC COMMUNICATION TECHNIQUE
- H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
- H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
- H04N23/70—Circuitry for compensating brightness variation in the scene
- H04N23/745—Detection of flicker frequency or suppression of flicker wherein the flicker is caused by illumination, e.g. due to fluorescent tube illumination or pulsed LED illumination
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/061—Sources
Abstract
The present invention relates to a kind of devices for observing microlayer model comprising: micro-fluidic chip has the microchannel for the microlayer model for including cycle movement；Flasher, the control signal provided according to a flasher control module generate the scintillation light with default scintillation parameter；And image capture module, acquire the image of the microlayer model in the microchannel of the micro-fluidic chip irradiated by the scintillation light.The present invention controls flasher by flasher control module and generates the scintillation light of specific frequency, and irradiates micro-fluidic chip；Subsequent picture acquisition module acquires the image on micro-fluidic chip；It can be obtained the information such as size, pattern and the flux of microlayer model by the frequency of these images and scintillation light.Relative to traditional the methods of high-speed camera and photoelectric tube, the present invention has the advantages that obtaining comprehensive, convenient, at low cost, the easy building of information, can be widely applied in miniaturization microfluidic system.
Description
Technical field
The present invention relates to a kind of device and method for observing microlayer model, more particularly to it is a kind of it is micro-fluidic in for it is high-throughput,
The device and method of the observation microlayer model of periodical microlayer model.
Background technique
Micro-fluidic chip (microfluidic chip) is also known as chip lab (lab on a chip), refers to raw
The basic operation units such as sample preparation, reaction, separation, detection involved in the fields such as object and chemistry are integrated or are integrated into substantially
On the chip of one piece several square centimeters (even more small), network is formed by microchannel, whole system is run through with controlled fluid, to
Replace a kind of technology of the various functions of standard biologic or chemical laboratory.It is developed so far, microfluidic chip technology has begun
It is applied in the different field of life science.Recently, the microflow control technique based on drop causes more and more concerns.With
Microchannel compares, and microlayer model has the advantage that smaller, and flux is higher, and mass-and heat-transfer is rapider, transport and fixed
It is more flexible, and a relatively stably and controllable drop microenvironment can be formed；Particularly, in micro-fluidic chip, people
A large amount of microlayer model can be generated in a short time.Therefore, in many high throughput analysis in chemistry and biology field, micro- liquid
Drop is used for the Large-scale Screening (such as protein crystal, DNA analysis) of reaction condition as reactor, or various for wrapping up
A large amount of particle (including cell, bacterium etc.) is to carry out various biological studies.
Towards these applications, need to construct the device or system based on drop micro-fluidic chip.In these device or systems
In, people generally require the information (size, pattern, movement speed, generation frequency etc.) for understanding microlayer model in real time.To realize this
Purpose, existing method and technique include:
(a) microlayer model is observed by using conventional ccd image sensor；The shortcomings that such method, is: due to the sampling of CCD
Frame speed is limited, therefore, it is difficult to directly observe the microlayer model of high-throughput (high-speed motion), and obtains the information of microlayer model.
(b) it is obtained by using the motion process of microlayer model in high-speed camera shooting microfluidic channel by image analysis
Take the relevant information of microlayer model；The shortcomings that such method, is: expensive, be difficult to integrate, it is difficult to the parameter of real-time measurement drop.
(c) optical signalling issued by using photodetection pipe detection microlayer model, to speculate the information of microlayer model；It is such
The shortcomings that method, is: being difficult to comprehensively obtain the information of microlayer model, is usually applicable only to the counting of drop.
Summary of the invention
In order to solve the above-mentioned problems of the prior art, the present invention is intended to provide a kind of device for observing microlayer model and side
Method easily to observe high-throughput microlayer model, the relevant information of Overall Acquisition microlayer model, and reduces cost.
A kind of device of observation microlayer model described in one of present invention comprising:
Micro-fluidic chip has the microchannel for the microlayer model for including cycle movement；
Flasher, the control signal provided according to a flasher control module, which generates, has default scintillation parameter
Scintillation light；And
Image capture module acquires the microlayer model in the microchannel of the micro-fluidic chip irradiated by the scintillation light
Image.
In the device of above-mentioned observation microlayer model, the micro-fluidic chip is using one in silicon, glass, quartz and polymer
Kind or a variety of be made.
In the device of above-mentioned observation microlayer model, the reproduced frequencies of the microlayer model are greater than 30Hz；The body of the microlayer model
Long-pending range is 0.06pL~4nL.
In the device of above-mentioned observation microlayer model, the scintillation parameter of the scintillation light includes the frequency, duty ratio and phase
Position, wherein the duty ratio of the scintillation light is continuously adjusted in the range of 0~100%, the phase of the scintillation light 0~
It is continuously adjusted in the range of 360 °.
In the device of above-mentioned observation microlayer model, the microchannel includes: the import and confession flowed into for the microlayer model
The outlet of the microlayer model outflow.
In the device of above-mentioned observation microlayer model, the flasher includes: to provide the LED array or xenon of the scintillation light
Lamp, and to the lens combination that the scintillation light is focused.
In the device of above-mentioned observation microlayer model, the response time of the flasher is less than 30ms.
In the device of above-mentioned observation microlayer model, the flasher control module includes: man-machine interactive interface and the people
Controller, the switch module being connect with the controller and the driving being connect with the switch module electricity of machine interactive interface connection
Source, wherein the controller receives the control instruction of periphery input by man-machine interactive interface, and is generated according to the control instruction
Square wave with predeterminated frequency, duty ratio and phase, to control the switch state of the switch module, the switch module according to
The square wave controls the on-off of the driving power, and exports the control signal to the flasher.
A kind of method of observation microlayer model described in the two of the present invention comprising following steps:
Step S1 provides the device of observation microlayer model as described above；
Step S2, by the flasher control module control the flasher generate frequency from 30Hz from as low as
The scintillation light of high consecutive variations, while the micro-fluidic chip irradiated by the scintillation light is acquired by described image acquisition module
Microchannel in microlayer model image, and record the frequency f for observing scintillation light when microlayer model is static twice in successionk、
fk+1, then according to the following formula (1), calculate the actual reproduction frequency f for obtaining microlayer modeldroplets:
Step S3 controls the flasher generation frequency by the flasher control module and is fixed as fflashSudden strain of a muscle
Bright light, while being acquired by described image acquisition module micro- in the microchannel of the micro-fluidic chip irradiated by the scintillation light
The image of drop, according to the image obtain microlayer model size and topographical information, and according to two adjacent microlayer models in the image it
Between distance L and following formula (2), calculate obtain microlayer model movement velocity Vdroplets:
Vdroplets=Lfdroplets(2)；
Step S4 adjusts the frequency that the flasher generates by the flasher control module and is fixed as fflashIt dodges
The phase of bright light, while the microchannel of the micro-fluidic chip irradiated by the acquisition of described image acquisition module by the scintillation light
The image of interior microlayer model obtains microlayer model in the size and topographical information of different moments according to the image.
In the method for above-mentioned observation microlayer model, the actual reproduction frequency f of the microlayer modeldropletsFor the frequency of scintillation light
Rate fflashIntegral multiple.
Due to using above-mentioned technical solution, the present invention controls flasher by flasher control module and produces
The scintillation light of raw specific frequency, and irradiate micro-fluidic chip；Subsequent picture acquisition module acquires the image on micro-fluidic chip；It is logical
The frequency for crossing these images and scintillation light can be obtained the information such as size, pattern and the flux of microlayer model.Relative to traditional height
The methods of fast video camera and photoelectric tube, the present invention has the advantages that comprehensive, convenient, at low cost, the easy building of information is obtained, it can be with
It is widely used in minimizing in microfluidic system.
Detailed description of the invention
Fig. 1 is a kind of structure principle chart of the device of observation microlayer model of the invention；
Fig. 2 is a kind of structural schematic diagram of the embodiment of the device of observation microlayer model of the invention；
Fig. 3 is the structural schematic diagram of micro-fluidic chip in a kind of device of observation microlayer model of the invention；
Fig. 4 is the structural schematic diagram of flasher in a kind of device of observation microlayer model of the invention；
Fig. 5 is the structural schematic diagram of flasher control module in a kind of device of observation microlayer model of the invention；
Fig. 6 (a)-(d) is a kind of schematic illustration for the device and method for observing microlayer model of the present invention；
Fig. 7 (a), (b) are the effect contrast figures using ordinary light source and flasher acquisition image.
Specific embodiment
With reference to the accompanying drawing, presently preferred embodiments of the present invention is provided, and is described in detail.
As shown in Figs. 1-5, one of present invention, i.e., a kind of device for observing microlayer model, comprising: image capture module 1, miniflow
Control chip 2, flasher 3 and the flasher control module 4 connecting with flasher 3；In the present embodiment, with
OlympusI X51 type microscope 100 is that platform will be micro- in the external image capture module 1 in the Image Acquisition port of microscope 100
Fluidic chip 2 is placed on the objective table 5 of microscope 100, and flasher 3 and flasher control module 4 is external micro-
The light source entrance of mirror 100.
Image capture module 1 can be using image capturing system (such as CCD of conventional low frame rate (10~100 frames are per second)
Deng), in the present embodiment, QImaging Retiga 2000R camera can be used as image capture module 1.
Micro-fluidic chip 2 can be by micro-fluidic conventional die material, such as: one of silicon, glass, quartz and polymer or
A variety of compositions can be used dimethyl silicone polymer (PDMS) and constitute (the master of this material of micro-fluidic chip 2 in the present embodiment
Wanting advantage is: cheap, simple processing)；Contain high-throughput (being greater than 30Hz), periodically fortune in the microchannel of the micro-fluidic chip 2
Dynamic microlayer model (size range of microlayer model is 0.06pL~4nL), in the present embodiment, the microchannel of micro-fluidic chip 2
Structure can use T-junction (T connector) structure 8, that is, the T-junction structure 8 includes: the oily phase to intersect vertically
Channel and water phase channel, wherein oil, which communicates, has oily phase import 6, and water phase channel has water phase import 7, and the oil phase channel
There is common outlet 9 with water phase channel.It should be noted that the structure of microchannel be also possible to other it is any have supply micro- liquid
Drip into import and for microlayer model outflow outlet channel design.
Flasher 3 includes: to provide the LED array 10 (being also possible to xenon lamp) of scintillation light and gather to the scintillation light
Burnt lens combination 11, in the present embodiment, flasher 3 are connect by interface 12 with the light source entrance of microscope 100, and are led to
It crosses cable 13 and connect with driving and control the flasher control module 4 of the flasher 3.
The controller that flasher control module 4 includes: man-machine interactive interface 14, is connect with the man-machine interactive interface 14
(for example, field programmable gate array (FPGA) controller 15 or digital processing unit DSP, microprocessor in the present embodiment
MCU etc.), the switch module (for example, solid-state relay 16 in the present embodiment) that is connect with the FPGA controller 15 and with this
The driving power (for example, LED drive power 17 in the present embodiment) that solid-state relay 16 connects, wherein FPGA controller 15
By man-machine interactive interface 14 receive periphery input control instruction, and according to the control instruction generate specific frequency (>=
30Hz), the square wave of duty ratio (0~100%), phase (0~360 °), to control the switch state of solid-state relay 16, solid-state
Relay 16 controls the on-off of LED drive power 17 according to the square wave (that is, control LED drive power 17 drives flasher 3
Whether), and corresponding control signal is exported to flasher 3.
It can be seen that in the apparatus of the present:
The control signal that flasher 3 is used to be exported according to flasher control module 4, which generates, has default scintillation parameter
(including frequency, phase and duty ratio etc.) and the scintillation light for being used to irradiate micro-fluidic chip 2, specifically, scintillation light irradiation is micro-
The optically transparent observation area of fluidic chip 2, and the response time of flasher is less than 30ms, to guarantee flasher energy
" quick " flashing, the image that otherwise will lead to the acquisition of image capture module 1 seriously trail, are fuzzy；
Image capture module 1 acquires the image of the microlayer model in the microchannel of the micro-fluidic chip 2 irradiated by scintillation light, with
The information such as size, pattern and the flux of microlayer model are obtained according to the image and scintillation parameter.
The two of the present invention, i.e., a kind of method for observing microlayer model, comprising the following steps:
Step S1 provides the device of observation microlayer model as described above；
It is continuous from low to high from 30Hz to control the generation frequency of flasher 3 by flasher control module 4 by step S2
The scintillation light (that is, the control signal that flasher control module 4 exports is the swept-frequency signal from 30Hz) of variation, passes through simultaneously
Image capture module 1 acquires the image of the microlayer model in the microchannel of the micro-fluidic chip 2 irradiated by the scintillation light, and the company of record
The continuous frequency f for observing scintillation light when microlayer model is static twicek、fk+1, then according to the following formula (1), calculate and obtain microlayer model
Actual reproduction frequency fdroplets:
Step S3 controls the generation of flasher 3 frequency by flasher control module 4 and is fixed as fflashScintillation light
(the actual reproduction frequency f of microlayer modeldropletsWith the frequency f of scintillation lightflashMeet condition: fdroplets=Nfflash, wherein N
=1,2,3 ...) in the microchannel for, while by image capture module 1 acquiring the micro-fluidic chip 2 irradiated by the scintillation light
Microlayer model image (at this time image be clearly, it is static), according to the image obtain microlayer model size and topographical information, and
According to the distance between two adjacent microlayer models L in the image and following formula (2), the movement velocity for obtaining microlayer model is calculated
Vdroplets:
Vdroplets=Lfdroplets(2)；
Step S4 adjusts the phase for the above-mentioned scintillation light that flasher 3 generates by flasher control module 4, simultaneously
The image that the microlayer model in the microchannel of the micro-fluidic chip 2 irradiated by the scintillation light is acquired by image capture module 1, according to
The image obtains microlayer model in the size and topographical information of different moments.
As shown in Fig. 6 (a)-(d), high-throughput microlayer model is usually what height reproduced in micro-fluidic chip 2, such as attached drawing 6
(a) shown in: the microlayer model in micro-fluidic chip 2 has very strong periodicity；For ease of description, herein successively by a drop
Period is divided into I, II, III, IV state, and in drop is micro-fluidic, this four states are circuited sequentially, and embodies the period of drop generation
Property.In order to make it easy to understand, the period equivalent signal of microlayer model, corresponding status indicator are illustrated in attached drawing 6 (b) using triangular wave
In triangular waveform.As shown in attached drawing 6 (c)-(d), as the actual reproduction frequency f of microlayer modeldropletsIt is the frequency of scintillation light
fflashIntegral multiple when, the image capture module 1 of low speed can collect the static image of II state at any time.Similarly,
By the phase for adjusting scintillation light, so that it may collect image corresponding to the different conditions of a drop period, pass through these
Image, so that it may obtain microlayer model in information such as size, the patterns of different moments.
The present invention is in implementation process, oily 6 oil inlet of phase import of micro-fluidic chip 2, and water phase import 7 is intake, and is applied simultaneously
Add the driving pressure of 50kPa.Under these conditions, micro-fluidic chip 2 can generate the microlayer model of 40 microns of diameter.According to this
The method of invention first carries out above-mentioned steps S2 (i.e. progress frequency sweep observation mode), the reproduced frequencies of practical calculated microlayer model
fdroplets=27Hz；Then step S3 (i.e. progress frequency locking observation mode), the frequency for the scintillation light that flasher 3 is generated are executed
fflashIt is set to 427Hz, image is acquired by QImaging Retiga 2000R camera, shown in image such as attached drawing 7 (b).Pass through figure
Size and topographical information as microlayer model can be observed.Meanwhile the distance L ≈ between two adjacent microlayer models can get according to this image
110 microns, and then obtain the speed V of microlayer modeldroplets≈47mm/s.Fig. 7 is shown under identical condition, uses normal light
Source (a) and use LED flasher (b), the image of acquisition.From can be seen that in Fig. 7 (a), (b) under ordinary light source, due to liquid
Drop movement is too fast, and the CCD of low speed defies capture clearly image, and the device of the invention and method then efficiently solve this and ask
Topic.
In conclusion the present invention has the advantages that convenient, at low cost, easy building, has good application value.
The device of the invention and method can be widely applied in miniaturization microfluidic system.
Above-described, only presently preferred embodiments of the present invention, the range being not intended to limit the invention, of the invention is upper
Stating embodiment can also make a variety of changes.Made by i.e. all claims applied according to the present invention and description
Simply, equivalent changes and modifications fall within the claims of the invention patent.The not detailed description of the present invention is
Routine techniques content.
Claims (9)
1. a kind of method for observing microlayer model, which is characterized in that the described method comprises the following steps:
Step S1, provides the device of an observation microlayer model, which includes:
Micro-fluidic chip has the microchannel for the microlayer model for including cycle movement；
Flasher, the control signal provided according to a flasher control module generate the flashing with default scintillation parameter
Light；And
Image capture module acquires the figure of the microlayer model in the microchannel of the micro-fluidic chip irradiated by the scintillation light
Picture；
Step S2 controls the flasher generation frequency by the flasher control module and connects from low to high from 30Hz
The scintillation light of continuous variation, while the micro- of the micro-fluidic chip irradiated by the scintillation light is acquired by described image acquisition module
The image of microlayer model in channel, and record the frequency f for observing scintillation light when microlayer model is static twice in successionk、fk+1, so
Afterwards according to the following formula (1), the reproduced frequencies f for obtaining microlayer model is calculateddroplets:
Step S3 controls the flasher generation frequency by the flasher control module and is fixed as fflashFlashing
Light, while micro- liquid in the microchannel of the micro-fluidic chip irradiated by the scintillation light by the acquisition of described image acquisition module
The image of drop obtains the size and topographical information of microlayer model according to the image, and according in the image between two adjacent microlayer models
Distance L and following formula (2), calculate obtain microlayer model movement velocity Vdroplets:
Vdroplets=Lfdroplets(2)；
Step S4 adjusts the frequency that the flasher generates by the flasher control module and is fixed as fflashScintillation light
Phase, while in the microchannel of the micro-fluidic chip irradiated by the scintillation light by the acquisition of described image acquisition module
The image of microlayer model obtains microlayer model in the size and topographical information of different moments according to the image.
2. the method for observation microlayer model according to claim 1, which is characterized in that the reproduced frequencies of the microlayer model
fdropletsFor the frequency f of scintillation lightflashIntegral multiple.
3. the method for observation microlayer model according to claim 1, which is characterized in that the micro-fluidic chip uses silicon, glass
One of glass, quartz and polymer a variety of are made.
4. the method for observation microlayer model according to claim 1, which is characterized in that the reproduced frequencies of the microlayer model are greater than
30Hz；The range of the volume of the microlayer model is 0.06pL~4nL.
5. the method for observation microlayer model according to claim 1, which is characterized in that the scintillation parameter of the scintillation light includes
Frequency, duty ratio and phase, wherein the duty ratio of the scintillation light is continuously adjusted in the range of 0~100%, the flashing
The phase of light is continuously adjusted in the range of 0~360 °.
6. the method for observation microlayer model according to claim 1, which is characterized in that the microchannel includes: for described micro-
The import that drop flows into and the outlet for microlayer model outflow.
7. the method for observation microlayer model according to claim 1, which is characterized in that the flasher includes: to provide institute
The LED array or xenon lamp of scintillation light are stated, and to the lens combination that the scintillation light is focused.
8. the method for observation microlayer model according to claim 1, which is characterized in that the response time of the flasher is small
In 30ms.
9. the method for observation microlayer model according to claim 1, which is characterized in that the flasher control module packet
Include: man-machine interactive interface, the controller being connect with the man-machine interactive interface, the switch module being connect with the controller and with this
The driving power of switch module connection, wherein the controller receives the control instruction of periphery input by man-machine interactive interface,
And the square wave with predeterminated frequency, duty ratio and phase is generated according to the control instruction, to control the switch of the switch module
State, the switch module controls the on-off of the driving power according to the square wave, and exports the control to the flasher
Signal processed.
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