CN110296963A - A kind of fluorescence detection device and fluorescence detection method - Google Patents
A kind of fluorescence detection device and fluorescence detection method Download PDFInfo
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- CN110296963A CN110296963A CN201910510843.9A CN201910510843A CN110296963A CN 110296963 A CN110296963 A CN 110296963A CN 201910510843 A CN201910510843 A CN 201910510843A CN 110296963 A CN110296963 A CN 110296963A
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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
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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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
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
Abstract
The invention belongs to biological electronic technology fields, mainly provide a kind of fluorescence detection device and fluorescence detection method, multiple sample droplets of injection are merged by transparent micro-flow control chip, obtain corresponding fusion drop, and the fusion drop is driven to optical signalling detection zone, then exciting light is exported to the fusion drop by the exciting light sources, so that the fusion drop generates corresponding fluorescence signal under the action of the exciting light, the fluorescence signal is received by fluorescent acceptor, and the fluorescence signal is converted into corresponding data-signal, by data processing system, signal and default treatment conditions export corresponding testing result based on the data, it solves traditional digital microcurrent-controlled platform and can not while the fluorescent molecule in the drop on electrode be excited and be acquired in vertical direction fluorescence signal The problem of.
Description
Technical field
The invention belongs to biological electronic technology field more particularly to a kind of fluorescence detection devices and fluorescence detection method.
Background technique
In Bioexperiment, fluorescence signal is very important result marking tools.It is excited from one end using the light of certain wavelength
Fluorescent molecule in drop obtains the fluorescence signal of specific wavelength in the other end with receiver, and whether judgment experiment result whereby
Meet expection.It is anti-at reservoir pre-add 4-5 times generally by the mode manually controlled on traditional digital microcurrent-controlled platform
The reaction solution answered extracts the liquid of single in experiment from reservoir with electrode, then using optical signal detection system to reaction
Liquid afterwards is detected.
However, traditional digital microcurrent-controlled platform generallys use lighttight metal electrode driving drop, can not exist simultaneously
The fluorescent molecule in the drop on electrode is excited and acquired in vertical direction fluorescence signal, greatly reduces fluorescence inspection
The efficiency of survey.
Summary of the invention
The purpose of the present invention is to provide a kind of fluorescence detection device and fluorescence detection methods, it is intended to solve traditional number
Microfluidic platform generallys use lighttight metal electrode driving drop, can not be simultaneously in vertical direction to the drop on electrode
In fluorescent molecule the problem of being excited and acquired fluorescence signal, greatly reducing the efficiency of fluorescence detection.
The embodiment of the present application provides a kind of fluorescence detection device, and the fluorescence detection device includes:
Exciting light sources；
Transparent micro-flow control chip, for by injection multiple sample droplets merge, obtain corresponding fusion drop, and
The fusion drop is driven to optical signalling detection zone, wherein the optical signalling detection zone is exported set on exciting light sources
Exciting light optical path on, the fusion drop generates corresponding fluorescence signal under the action of the exciting light；
Fluorescent acceptor is converted to corresponding data-signal for receiving the fluorescence signal, and by the fluorescence signal；
And
Data processing module is connect with the fluorescent acceptor, and the data processing module for believing based on the data
Number and default treatment conditions export corresponding testing result.
Optionally, the transparent micro-flow control chip includes the transparent bottom crown and transparent top crown being stacked, wherein institute
It states transparent bottom crown surface and is formed with electrode layer, the transparent top crown surface is formed with reservoir, and the reservoir is set to institute
It states between transparent bottom crown and the transparent top crown；
The electrode layer drives the sample droplets in the reservoir for driving signal based on the received.
Optionally, the fluorescence detection device further include:
The volume of the sample droplets in the reservoir is adjusted in peristaltic pump system, the instruction for being inputted according to user
Section.
Optionally, the peristaltic pump system includes the peristaltic pump, pump line and aspiration needle sequentially connected；
The pump line is used to store the sample droplets of preset vol；
The peristaltic pump generates corresponding air pressure according to the instruction that user inputs, to pass through the aspiration needle to the liquid storage
The volume of sample droplets in slot is adjusted.
Optionally, the fluorescence detection device further include: host computer, microcontroller, relay module and power drives
Module；
The host computer is used to obtain the control ginseng of the transparent micro-flow control chip based on user's operation editor's powering order
Number；
The microcontroller is used to the control parameter being converted to corresponding level signal；
The relay module is used for the controlled terminal when the level signal is applied to control terminal and is closed, so that power drives
Module, the controlled terminal of the relay module, the transparent micro-flow control chip form electrical circuit；
The electric power driving module is used to when the controlled terminal of relay module closure be the transparent micro-fluidic core
Piece provides driving signal, to drive the drop on the transparent micro-flow control chip mobile.
Optionally, the host computer is also used to control the operating voltage and switching-on and switching-off state of the peristaltic pump system.
The embodiment of the present application also provides a kind of fluorescence detection method, the fluorescence detection method includes:
Multiple sample droplets of injection are merged using transparent micro-flow control chip, obtain corresponding fusion drop, and
The fusion drop is driven to optical signalling detection zone；Wherein, the optical signalling detection zone is set to the exciting light sources
In the optical path of the exciting light of output, the fusion drop generates corresponding fluorescence signal under the action of the exciting light；
The fluorescence signal is received by fluorescent acceptor, and the fluorescence signal is converted into corresponding data-signal；
By data processing module, signal and default treatment conditions export corresponding testing result based on the data.
Optionally, the fluorescence detection method further include:
The instruction inputted by peristaltic pump system according to user is to the sample liquid in the reservoir on transparent micro-flow control chip
The volume of drop is adjusted.
Optionally, the fluorescence detection method further include:
The control parameter of the transparent micro-flow control chip is obtained based on user's operation editor's powering order by host computer；
The control parameter is converted into corresponding level signal by microcontroller；
By relay module, when the level signal is applied to control terminal, controlled terminal is closed, so that the power drives
Module, the controlled terminal of relay module, transparent micro-flow control chip form electrical circuit；
It is mentioned by electric power driving module in the controlled terminal closure of the relay module for the transparent micro-flow control chip
For driving signal, to drive the drop on the transparent micro-flow control chip mobile.
Optionally, the fluorescence detection method further include:
It is adjusted by operating voltage and switching-on and switching-off state of the host computer to the peristaltic pump system.
The embodiment of the present application provides a kind of fluorescence detection device and fluorescence detection method, passes through transparent micro-flow control chip root
Multiple sample droplets of injection are merged according to the driving signal of input, obtain corresponding fusion drop, and by the fusion
Drop drives to optical signalling detection zone, then exports exciting light to the fusion drop by the exciting light sources, obtains
The received fluorescence signal of fluorescent acceptor is converted to corresponding data by photo-translating system and believed by corresponding fluorescence signal
Number, by data processing system, signal and default treatment conditions export corresponding testing result based on the data, solve
Traditional digital microcurrent-controlled platform can not simultaneously in vertical direction to the fluorescent molecule in the drop on electrode carry out excitation and
The problem of acquiring fluorescence signal.
Detailed description of the invention
Fig. 1 is the structural schematic diagram for the fluorescence detection device that one embodiment of the application provides.
Fig. 2 is the flow diagram for the fluorescence detection method that one embodiment of the application provides.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to the accompanying drawings and embodiments, right
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.
Fig. 1 is the structural schematic diagram for the fluorescence detection device that one embodiment of the application provides, shown in Figure 1, this
Application embodiment provides a kind of fluorescence detection device, and the fluorescence detection device includes:
Exciting light sources 10；
Transparent micro-flow control chip 20, for by injection multiple sample droplets merge, obtain corresponding fusion drop,
And the fusion drop is driven to optical signalling detection zone, wherein the optical signalling detection zone is set to the excitation light
In the optical path for the exciting light that source 10 exports, the fusion drop generates corresponding fluorescence signal under the action of the exciting light；
The fluorescence signal for receiving the fluorescence signal, and is converted to corresponding data and believed by fluorescent acceptor 30
Number；And
Data processing module 40 is connect with the fluorescent acceptor 30, and the data processing module 40 is used for based on described
Data-signal and default treatment conditions export corresponding testing result.
In the present embodiment, by by transparent micro-flow control chip 20 be set to exciting light sources 10 and fluorescent acceptor 30 it
Between, and the optical signalling detection zone being arranged on transparent micro-flow control chip 20 is made to be located at the exciting light of the transmitting of exciting light sources 10
In optical path so that transparent micro-flow control chip 20 will merge drop drive to optical signalling detection zone after can directly open it is sharp
10 pairs of fusion drops of illuminating source are irradiated, and are excited under exciting light irradiation by the reception of fluorescent acceptor 30 fusion drop
The fluorescence signal generated is sent out, and the fluorescence signal is converted into corresponding data-signal, finally by 40 base of data processing module
Corresponding testing result is exported in the data-signal and default treatment conditions, which can be according to user's
Detection needs to be arranged, for example, using liposome by the application test for thering is the plasmid of green fluorescent protein sequence to be transferred to cell,
Multiple sample droplets include the drop of celliferous drop, the drop comprising the liposome containing plasmid and cell culture medium,
Above-mentioned three kinds of sample droplets are passed through to the different sample holes injections being arranged on transparent micro-flow control chip 20 respectively, it is transparent micro-fluidic
Driving signal will wrap celliferous drop to chip 20 based on the received, the drop comprising the liposome containing plasmid merges,
Corresponding fusion drop is obtained, and fusion drop is moved to optical signalling detection zone, is then turned on the irradiation of exciting light sources 10
Then bluish violet exciting light receives fusion drop by fluorescent acceptor 30 and is stimulated the fluorescence signal of generation, the default processing
Condition is that the data-signal that the fluorescence signal is converted to is compared with green florescent signal, to judge green fluorescent protein
Whether expressed in cell, to further determine that plasmid, whether Successful transfection is into cell, i.e. the default treatment conditions can be with
For the pre-set comparison fluorescence signal of user, believe so that the received fluorescence signal of fluorescent acceptor is converted to corresponding data
Number to be compared with preset comparison fluorescence signal, whether succeeded with judgment experiment.
In one embodiment, transparent micro-flow control chip 20 can also be according to the driving signal that user inputs to fusion drop
Separating treatment is carried out, so that preset sample droplets are isolated from fusion drop, for example, transparent micro-flow control chip 20 is according to connecing
The driving signal of receipts will wrap celliferous drop, the drop comprising the liposome containing plasmid merges, and obtain corresponding
The drop of Fresh cell culture medium constantly can be merged drop with first and merged by one fusion drop after cell is adherent
The second fusion drop is obtained, and isolates the identical culture medium drop of a volume from the second fusion drop simultaneously, to protect
Card cell possesses fresh culture medium, reaches the purpose of continuity culture.
In one embodiment, the fluorescence detection device in the present embodiment can also include temperature control system, the temperature control system
It is adjusted for the environment temperature to transparent micro-flow control chip 20, so that transparent micro-flow control chip 20 can be completed at the same time
The culture of continuous cultivation and detection process of bacterium can be completed by the default treatment conditions in setting data processing module 40
Diversity detection to bacterium drop, for example, can be directly to culture to optical signalling detection zone by driving bacterium drop
The drop of bacterium carries out absorbance detection, and to judge the density of bacterium generation, the growing state of bacterium is judged by fluorescence brief introduction
Or gene expression dose, it can also be by the drug resistance of the absorbance detection tested bacteria of bacterium, specifically, will contain to be measured thin
The drop of bacterium and drop containing different pharmaceutical are injected from multiple sample delivery points, by providing corresponding drive to transparent micro-flow control chip 20
Dynamic signal moves these drops after being sufficiently mixed the drop of each drug containing and a drop containing tested bacteria
Different optical signalling detection zones is planted, the 10 intermittent emitting ultraviolet light of exciting light sources from one end, the fluorescence of the other end are passed through
Receiver 30 carries out the reception of ultraviolet light, so that the absorbance of tested bacteria drop is calculated by data processing module 40, thus
It derives the density of bacterium in drop, further judges whether bacterium is resistant to certain drug.Relative to existing artificial reality
It tests and compares, repeated strong and a large amount of work can be subjected to automation control using the fluorescence detection device in the present embodiment,
The labour of experimenter is greatly liberated, it can also be than being manually more quickly completed Resistance detection test.In addition, and in the market
The existing instrument that can automate progress Resistance detection is compared, the single experiment of the fluorescence detection device in this present embodiment
Amount of reagent is very low, and can increase the drug that can be detected simultaneously by increasing sample delivery point.
In one embodiment, the transparent micro-flow control chip 20 includes the transparent bottom crown being stacked and transparent upper pole
Plate, wherein the transparent bottom crown surface is formed with electrode layer, and the transparent top crown surface is formed with reservoir, the storage
Liquid bath is set between the transparent bottom crown and the transparent top crown；
The electrode layer drives the sample droplets in the reservoir for driving signal based on the received.
In the present embodiment, transparent micro-flow control chip 20 includes the transparent bottom crown and transparent top crown being stacked,
In, electrode layer is formed in the upper surface of bottom crown, and electrode layer includes multiple electrodes, and for receiving driving signal, each electrode is equal
It is covered by dielectric material and hydrophobic material, dielectric material plays the role of electric insulation, and electrowetting power is provided when electrode is powered
It is mobile with dielectrophoretic force driving drop.Top crown is by transparent conductive material (for example, indium oxide tin glass or FTO glass) structure
At the surface of top crown and bottom crown is coated with super hydrophobic material, for example, Teflon etc..In cold situation, electrode
Surface can not soak, and by applying driving voltage to electrode, electrowetting can occur for the electrode surface being energized, and drop is in electrode electricity
It is moved under the action of wetting.
In the present embodiment, the droplet size on transparent micro-flow control chip 20 can be adjusted by the way that number of poles is arranged
It is whole, that is to say, that driving voltage can be applied simultaneously for multiple adjacent electrodes, so that they drive one big drop mobile,
Or single electrode applies driving voltage and it is made to drive a droplet mobile.The size of big drop and droplet is controllable,
Big drop is separated into droplet, and droplet can be mixed into big drop.
In one embodiment, the electrode layer of the upper surface of bottom crown can be obtained by way of wet etching, first
The even one layer of positive photoresist of sol evenning machine is used on the ITO coating of ito glass, and heat fixation is carried out to the positive photoresist, it is then sharp
Preset electrode pattern is obtained with uv-exposure, after washing away extra photoresist with developer solution, by ito glass as 2- in etching solution
It is etched within 5 minutes.After etching, ito glass is cleaned up using deionized water, washes away remaining positive photoresist, thus
Bottom crown surface is formed with the electrode layer of default electrode pattern.
In one embodiment, ito glass could alternatively be FTO glass in above-described embodiment, and ITO coating replaces with FTO
The fluorine-doped tin oxide coating of glass surface.
In one embodiment, multiple reservoirs are provided on upper machine plate, multiple reservoirs correspond to multiple sample holes, user
It can according to need and the droplet size in corresponding reservoir is adjusted by sample holes.
In one embodiment, the fluorescence detection device further include:
The volume of the sample droplets in the reservoir is adjusted in peristaltic pump system, the instruction for being inputted according to user
Section.
In the present embodiment, user can be by peristaltic pump system in the reservoir being arranged on transparent micro-flow control chip 20
The volumes of sample droplets be adjusted, for example, according to the pressure difference of the instruction control peristaltic pump system of user's input, thus right
The volume of sample droplets is accurately controlled, further, can also be according to the instruction that user inputs in peristaltic pump system
Positive and negative anodes are configured to adjust the direction of rotation of peristaltic pump, reach the extra sample droplets in reservoir drawing back reagent
The purpose of pipe not only saves reagent cost, and also avoids decompression pressurization gas pump and valve required for traditional air pump,
Save the occupied space of liquid nitrogen bottle.
In one embodiment, the instruction of user's input can also include pump line switching command, by peristaltic pump system
In the pump lines of different inner diameters switch over, achieve the purpose that the flow of the sample droplets exported to pump line is adjusted, at this
In embodiment, which can be silicone tube, pass through high temperature resistant, low temperature resistant, oil resistivity and the good life using silicone tube
Stability is managed, error caused by can deforming during the work time due to pump line to avoid peristaltic pump system.In one embodiment,
The peristaltic pump system includes the peristaltic pump, pump line and aspiration needle sequentially connected；
The pump line is used to store the sample droplets of preset vol；
The peristaltic pump generates corresponding air pressure according to the instruction that user inputs, to pass through the aspiration needle to the liquid storage
The volume of sample droplets in slot is adjusted.
In the present embodiment, the storage of sample droplets is carried out by pump line, peristaltic pump is carried out according to the instruction that user inputs
Rotation, to generate corresponding air pressure, to drive aspiration needle that the volume of the sample droplets in reservoir is adjusted, specifically
, peristaltic pump generates corresponding air pressure and aspirates to the air column in pump line, thus the movement to the sample droplets in pump line
Direction is controlled.
In one embodiment, the aspiration needle in the present embodiment and reservoir are tightly connected, and can be filled to avoid fluorescence detection
Set the sample introduction error generated due to shake.
In one embodiment, a peristaltic pump can access more pump lines simultaneously, more pump lines respectively with multiple suctions
Needle corresponds, and can store different types of sample droplets in multiple pump lines, and user can according to need needed for being stored with
The pump line of sample droplets accesses peristaltic pump simultaneously, and the effect of multiple sample droplets while input may be implemented by opening peristaltic pump
Fruit not only saves the number of peristaltic pump, can also inject a variety of required sample droplets simultaneously, realizes and injects to sample droplets
Be precisely controlled, passing through, which reduces time difference of different sample droplets injection, eliminates experimental error.
In one embodiment, the pump line in the present embodiment may include the different silicone tube of multiple internal diameters, to pass through
Switching silicone tube realization the type of sample droplets and the output flow of sample droplets are controlled, reach to sample-adding process into
The purpose that is precisely controlled of row, further, the tail portion of silicone tube can be set to half-conical, be conducive to silicone tube and aspiration needle it
Between connection.In one embodiment, the peristaltic pump system further includes drop memory, and one end of peristaltic pump is deposited from drop
Sample droplets are sucked in reservoir, and the sample droplets are exported from the other end into pump line, so as to by storing to drop
Device is loaded, and achievees the purpose that endlessly to provide sample droplets for pump line, and realization is carried out continuously sample culture and glimmering
The effect of light detection.
In the present embodiment, the other end of peristaltic pump uses the structure of pump line and aspiration needle, and aspiration needle can be reached vertically
Near chip surface, sample droplets are allowed to get by peristaltic pump pressurization, sample droplets are sticked to the electrode of drop reaction zone after getting
Except reservoir, the reservoir can be preset reservoir electrode so that the drop of microlitre rank magnitude is able to respond
The dielectrophoresis effect that bright micro-fluidic chip 20 generates, specifically, since transparent micro-flow control chip 20 uses super-drainage structure, nothing
Method is added in chip surface with the mode of similar pipette tips.Liquid storage rooved face can be allowed to become hydrophilic surface by opening reservoir electrode, this
The trace sample drop that aspiration needle is got can be successfully injected into transparent micro-flow control chip 20 by sample.
In one embodiment, the aspiration needle in the present embodiment can use injection needle, have half on the injection needle
The diameter difference of spherical structure, further, the peristaltic pump system in the present embodiment can also include the fixture for fixing aspiration needle
Structure, which can simultaneously be fixed multiple aspiration needles, to avoid aspiration needle in the mistake of injecting sample drop
Shake occurs in journey and generates error.
Further, the clamp structure in the present embodiment can be also used for multiple fixed pump lines, avoid pump line in peristaltic pump
It is generated when work and shakes the injection generation error for leading to sample droplets.
In the present embodiment, sample droplets flow to injection needle by silicone tube, are made by the beveled cone of injection needle tip
It obtains sample droplets to become larger, then applies the reservoir being on the transparent top crown of injection needle contact, then by transparent micro-
Fluidic chip driving sample droplets enter the reaction zone of transparent micro-flow control chip, so that sample introduction is completed, further, when user needs
When extracting sample droplets out, transparent micro-flow control chip driving sample droplets enter reservoir, are then generated by adjusting peristaltic pump
Negative pressure, to exporting sample droplets into pump line.
In one embodiment, the fluorescence detection device further include: host computer, microcontroller, relay module and
Electric power driving module；
The host computer is used to obtain the control of the transparent micro-flow control chip 20 based on user's operation editor's powering order
Parameter；
The microcontroller is used to the control parameter being converted to corresponding level signal；
The relay module is used for the controlled terminal when the level signal is applied to control terminal and is closed, so that the power supply
Drive module, the controlled terminal of the relay module, the transparent micro-flow control chip 20 form electrical circuit；
The electric power driving module is used to when the controlled terminal of relay module closure be the transparent micro-fluidic core
Piece 20 provides driving signal, to drive the drop on the transparent micro-flow control chip 20 mobile.
In the present embodiment, host computer, microcontroller, relay module, electric power driving module and transparent micro-fluidic core
Piece 20 forms digital microfluidic system, specifically, the controlled terminal of relay module passes through spring needle and transparent micro-flow control chip 20
On multiple electrodes be electrically connected, each electrode includes electrode sequence number, for example, the number of 20 top electrode of transparent micro-flow control chip
It can be 92,94 or 96.Relay module includes multiple relay switches, the control terminal of each relay switch with
Microcontroller electrical connection, the controlled terminal of each relay switch be electrically connected with electric power driving module and pass through spring needle with it is transparent
Electrode electrical connection on micro-fluidic chip 20.Host computer is used to obtain transparent miniflow based on user's operation editor powering order
The control parameter of chip 20 is controlled, and control parameter is sent to microcontroller, wherein the control parameter may include target electrode
Serial number, single step power-up time and single step interval time.Therefore, host computer is also used to connect is obtained based on user's operation editor's powering order
To the single step power-up time and single step interval time of target electrode serial number and target electrode, and it is sent to microcontroller.
In the present embodiment, host computer operation has the code compilation software for drop driving, which can
Will be carried out based on target electrode serial number, single step power-up time and the single step interval time that user's operation editor's powering order obtains
Compiling is obtained comprising powering order, target electrode serial number, single step power-up time and the code of single step interval time and with TXT text
The storage of part form.In host computer operation, host computer calls this to be stored with the TXT file of code, suitable according to the power-up in code
Sequence, target electrode serial number, single step power-up time and single step interval time realize that the closing of relay switch in relay module is suitable
Sequence and interval, to send corresponding driving signal to transparent micro-flow control chip 20.
In one embodiment, host computer is also used to exciting light sources 10, fluorescent acceptor 30 and data processing mould
The Systems for optical inspection that block 40 forms is controlled, thus on realizing Systems for optical inspection and transparent micro-flow control chip 20 simultaneously
Electricity, and exciting light sources 10 are opened simultaneously when transparent micro-flow control chip 20 drives fusion drop to optical signalling detection zone, it keeps away
Exempt from because of the error that detection delay generates.
In one embodiment, the host computer is also used to control the operating voltage of the peristaltic pump system and power supply is opened
Off status.
In the present embodiment, host computer is also used to control peristaltic pump system, specifically, host computer can be by right
The operating voltage and switching-on and switching-off state of peristaltic pump system are adjusted, so that the working condition to peristaltic pump system is adjusted
Section, for example, controlling whether peristaltic pump system starts by the switching-on and switching-off state of control peristaltic pump system, by controlling peristaltic pump
The operating voltage of system is to control the velocity of rotation of peristaltic pump, to complete to control the flow velocity of sample droplets, reaches pair
The purpose that the volume of sample droplets in reservoir is adjusted.
In one embodiment, host computer can be personal computer, such as desktop computer or laptop, meanwhile,
Equipped with Labview software or the EXE program file being set in fpga chip on host computer.
In one embodiment, host computer can also be deployed with human-computer interaction interface, and human-computer interaction interface is provided to saturating
The automatic operation and Stateful Inspection function of bright micro-fluidic chip 20, user can pass through the human-computer interaction interface selection target electricity
Pole can also be called the TXT file for being stored with code by the human-computer interaction interface, the code in TXT file is sent to micro-
Controller, so that transparent micro-flow control chip 20 is according to powering order specified in code, target electrode serial number, single step power-up time
The closing sequence of relay switch and interval in relay module are realized with single step interval time.In addition, in transparent micro-fluidic core
When piece 20 is run, user can also monitor in real time the on off operating mode of each electrode, conduction time and each by human-computer interaction interface
The executive condition of a step.
In one embodiment, digital microfluidic system further includes magnetic module, magnetic module and Arduino communication link
It connects, transparent micro-flow control chip 20 is set at the magnetic force position of magnetic module, and magnetic module is used for the wireless remote control in Arduino
The lower magnetic bead by transparent micro-flow control chip 20 is separated and is mixed with sample droplets.Magnetic bead due to it is big with surface area,
Stable chemical performance, surface in modifying other molecules, being easy to the advantages that manipulating in externally-applied magnetic field, (are such as exempted from biochemical analysis
Epidemic disease reaction, detection of nucleic acids, protein enrichment) cell separation etc. application in have huge application value, in digital microfluidic system
Drop and magnetic bead are combined, can be further improved the flux of reaction and shorten the reaction time.Such as pass through chemical modification
Antibody or DNA chain are connected on magnetic bead, target molecule or cell will form specific interaction between magnetic bead.And
Under the action of externally-applied magnetic field i.e. magnetic module, the target molecule or cell of capture are easy to be eluted, later for next
The reaction or analysis of step.I.e. pass through magnetic module adsorb magnetic bead and using driving voltage drive drop, can be achieved with drop with
The separation of magnetic bead, and in the case where magnetic module is not involved in, drop is driven merely with driving voltage, can be achieved with drop and magnetic
The mixing of pearl.
Fig. 2 is the flow diagram for the fluorescence detection method that one embodiment of the application provides, shown in Figure 2, this
Fluorescence detection method in embodiment includes:
Step S10: multiple sample droplets of injection are merged using transparent micro-flow control chip 20, obtain corresponding melt
Drop is closed, and the fusion drop is driven to optical signalling detection zone；Wherein, the optical signalling detection zone is set to described sharp
In the optical path for the exciting light that illuminating source 10 exports, the fusion drop generates corresponding fluorescence under the action of the exciting light
Signal；
Step S20: receiving the fluorescence signal by fluorescent acceptor 30, and the fluorescence signal is converted to corresponding
Data-signal；
Step S30: by data processing module 40, signal and the output of default treatment conditions are corresponding based on the data
Testing result.
In the present embodiment, by by transparent micro-flow control chip 20 be set to exciting light sources 10 and fluorescent acceptor 30 it
Between, and the optical signalling detection zone being arranged on transparent micro-flow control chip 20 is made to be located at the exciting light of the transmitting of exciting light sources 10
In optical path so that transparent micro-flow control chip 20 will merge drop drive to optical signalling detection zone after can directly open it is sharp
10 pairs of fusion drops of illuminating source are irradiated, and are excited under exciting light irradiation by the reception of fluorescent acceptor 30 fusion drop
The fluorescence signal generated is sent out, and the fluorescence signal is converted into corresponding data-signal, finally by 40 base of data processing module
Corresponding testing result is exported in the data-signal and default treatment conditions, which can be according to user's need
Be arranged, for example, using liposome by the application test for thering is the plasmid of green fluorescent protein sequence to be transferred to cell, multiple samples
This drop includes the drop of celliferous drop, the drop comprising the liposome containing plasmid and cell culture medium, will be above-mentioned
Three kinds of sample droplets pass through the different sample holes injections being arranged on transparent micro-flow control chip 20, transparent micro-flow control chip 20 respectively
Driving signal will wrap celliferous drop based on the received, the drop comprising the liposome containing plasmid merges, and obtain pair
The fusion drop answered, and fusion drop is moved to optical signalling detection zone, it is then turned on exciting light sources 10 and irradiates bluish violet
Exciting light, then receives fusion drop by fluorescent acceptor 30 and is stimulated the fluorescence signal of generation, which is
The data-signal that the fluorescence signal is converted to is compared with green florescent signal, with judge green fluorescent protein whether
It is expressed in cell, to further determine that plasmid, whether Successful transfection is into cell.
In one embodiment, transparent micro-flow control chip 20 can also be according to the driving signal that user inputs to fusion drop
Separating treatment is carried out, so that preset sample droplets are isolated from fusion drop, for example, transparent micro-flow control chip 20 is according to connecing
The driving signal of receipts will wrap celliferous drop, the drop comprising the liposome containing plasmid merges, and obtain corresponding
The drop of Fresh cell culture medium constantly can be merged drop with first and merged by one fusion drop after cell is adherent
The second fusion drop is obtained, and isolates the identical culture medium drop of a volume from the second fusion drop simultaneously, to protect
Card cell possesses fresh culture medium, reaches the purpose of continuity culture.
In one embodiment, the fluorescence detection method further include:
Step S11: the instruction inputted by peristaltic pump system storage according to user is to the liquid storage on transparent micro-flow control chip 20
The volume of sample droplets in slot is adjusted.
In one embodiment, the fluorescence detection method further include:
Step S12: the control of the transparent micro-flow control chip is obtained based on user's operation editor's powering order by host computer
Parameter processed；
Step S13: the control parameter is converted to by corresponding level signal by microcontroller；
Step S14: by relay module, when the level signal is applied to control terminal, controlled terminal is closed, so that described
Electric power driving module, the controlled terminal of relay module, transparent micro-flow control chip 20 form electrical circuit；
Step S15: being the transparent miniflow when the controlled terminal of the relay module is closed by electric power driving module
It controls chip 20 and driving signal is provided, to drive the drop on the transparent micro-flow control chip 20 mobile.
In the present embodiment, host computer, microcontroller, relay module, electric power driving module and transparent micro-fluidic core
Piece 20 forms digital microfluidic system, specifically, the controlled terminal of relay module passes through spring needle and transparent micro-flow control chip 20
On multiple electrodes be electrically connected, each electrode includes electrode sequence number, for example, the number of 20 top electrode of transparent micro-flow control chip
It can be 92,94 or 96.Relay module includes multiple relay switches, the control terminal of each relay switch with
Microcontroller electrical connection, the controlled terminal of each relay switch be electrically connected with electric power driving module and pass through spring needle with it is transparent
Electrode electrical connection on micro-fluidic chip 20.Host computer is used to obtain transparent miniflow based on user's operation editor powering order
The control parameter of chip 20 is controlled, and control parameter is sent to microcontroller, wherein the control parameter may include target electrode
Serial number, single step power-up time and single step interval time.Therefore, host computer is also used to connect is obtained based on user's operation editor's powering order
To the single step power-up time and single step interval time of target electrode serial number and target electrode, and it is sent to microcontroller.
In one embodiment, the fluorescence detection method further include:
Step S111: it is adjusted by operating voltage and switching-on and switching-off state of the host computer to the peristaltic pump system
Section.
In the present embodiment, host computer can pass through the operating voltage and switching-on and switching-off state progress to peristaltic pump system
It adjusts, so that the working condition to peristaltic pump system is adjusted, for example, passing through the switching-on and switching-off state of control peristaltic pump system
Whether control peristaltic pump system starts, and the velocity of rotation of peristaltic pump is controlled by the operating voltage of control peristaltic pump system, from
And complete to control the flow velocity of sample droplets, achieve the purpose that the volume to the sample droplets in reservoir is adjusted.
The embodiment of the present application provides a kind of fluorescence detection device and fluorescence detection method, passes through transparent micro-flow control chip 20
Multiple sample droplets of injection are merged, obtain corresponding fusion drop, and the fusion drop is driven to optics and is believed
Number detection zone, then by the exciting light sources 10 to fusion drop output exciting light, so that the fusion drop exists
Corresponding fluorescence signal is generated under the action of the exciting light, the fluorescence signal is received by fluorescent acceptor 30, and by institute
It states fluorescence signal and is converted to corresponding data-signal, pass through data processing system signal and default processing item based on the data
Part exports corresponding testing result, solve traditional digital microcurrent-controlled platform can not and meanwhile in vertical direction on electrode
The problem of fluorescent molecule in drop is excited and acquires fluorescence signal.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (10)
1. a kind of fluorescence detection device, which is characterized in that the fluorescence detection device includes:
Exciting light sources；
Transparent micro-flow control chip obtains corresponding fusion drop for merging multiple sample droplets of injection, and by institute
It states fusion drop to drive to optical signalling detection zone, wherein the optical signalling detection zone is exported set on the exciting light sources
Exciting light optical path on, the fusion drop generates corresponding fluorescence signal under the action of the exciting light；
Fluorescent acceptor is converted to corresponding data-signal for receiving the fluorescence signal, and by the fluorescence signal；And
Data processing module is connect with the fluorescent acceptor, the data processing module for signal based on the data with
And default treatment conditions export corresponding testing result.
2. fluorescence detection device as described in claim 1, which is characterized in that the transparent micro-flow control chip includes being stacked
Transparent bottom crown and transparent top crown, wherein the transparent bottom crown surface is formed with electrode layer, the transparent top crown table
Face is formed with reservoir, and the reservoir is set between the transparent bottom crown and the transparent top crown；
The electrode layer drives the sample droplets in the reservoir for driving signal based on the received.
3. fluorescence detection device as claimed in claim 2, which is characterized in that the fluorescence detection device further include:
The volume of the sample droplets in the reservoir is adjusted in peristaltic pump system, the instruction for being inputted according to user.
4. fluorescence detection device as claimed in claim 3, which is characterized in that the peristaltic pump system include sequentially connect it is compacted
Dynamic pump, pump line and aspiration needle；
The pump line is used to store the sample droplets of preset vol；
The peristaltic pump generates corresponding air pressure according to the instruction that user inputs, to pass through the aspiration needle in the reservoir
The volumes of sample droplets be adjusted.
5. fluorescence detection device as claimed in claim 4, which is characterized in that the fluorescence detection device further include: host computer,
Microcontroller, relay module and electric power driving module；
The host computer is used to obtain the control parameter of the transparent micro-flow control chip based on user's operation editor's powering order；
The microcontroller is used to the control parameter being converted to corresponding level signal；
The relay module is used for the controlled terminal when the level signal is applied to control terminal and is closed, so that the power drives
Module, the controlled terminal of the relay module, the transparent micro-flow control chip form electrical circuit；
The electric power driving module is used to mention when the controlled terminal of relay module closure for the transparent micro-flow control chip
For driving signal, to drive the drop on the transparent micro-flow control chip mobile.
6. fluorescence detection device as claimed in claim 5, which is characterized in that the host computer is also used to control the peristaltic pump
The operating voltage and switching-on and switching-off state of system.
7. a kind of fluorescence detection method, which is characterized in that the fluorescence detection method includes:
Multiple sample droplets of injection are merged using transparent micro-flow control chip, obtain corresponding fusion drop, and by institute
Fusion drop is stated to drive to optical signalling detection zone；Wherein, the optical signalling detection zone is set to swashing for exciting light sources output
In luminous optical path, the fusion drop generates corresponding fluorescence signal under the action of the exciting light；
The fluorescence signal is received by fluorescent acceptor, and the fluorescence signal is converted into corresponding data-signal；
By data processing module, signal and default treatment conditions export corresponding testing result based on the data.
8. fluorescence detection method as claimed in claim 7, which is characterized in that the fluorescence detection method further include:
The instruction inputted by peristaltic pump system according to user is to the sample droplets in the reservoir on transparent micro-flow control chip
Volume is adjusted.
9. fluorescence detection method as claimed in claim 7, which is characterized in that the fluorescence detection method further include:
The control parameter of the transparent micro-flow control chip is obtained based on user's operation editor's powering order by host computer；
The control parameter is converted into corresponding level signal by microcontroller；
By relay module when the level signal is applied to control terminal controlled terminal be closed so that electric power driving module, after
Controlled terminal, the transparent micro-flow control chip of electrical appliance module form electrical circuit；
Drive is provided for the transparent micro-flow control chip in the controlled terminal closure of the relay module by electric power driving module
Dynamic signal, to drive the drop on the transparent micro-flow control chip mobile.
10. fluorescence detection method as claimed in claim 8, which is characterized in that the fluorescence detection method further include:
It is adjusted by operating voltage and switching-on and switching-off state of the host computer to the peristaltic pump system.
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