CN108020490A - A kind of high flux screening equipment using drop micro-fluidic chip - Google Patents
A kind of high flux screening equipment using drop micro-fluidic chip Download PDFInfo
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- CN108020490A CN108020490A CN201710486937.8A CN201710486937A CN108020490A CN 108020490 A CN108020490 A CN 108020490A CN 201710486937 A CN201710486937 A CN 201710486937A CN 108020490 A CN108020490 A CN 108020490A
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- fluidic chip
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- 238000012216 screening Methods 0.000 title claims abstract description 140
- 230000004907 flux Effects 0.000 title claims abstract description 33
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- 238000001917 fluorescence detection Methods 0.000 claims abstract description 29
- 230000005284 excitation Effects 0.000 claims abstract description 14
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- 238000004519 manufacturing process Methods 0.000 description 5
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- 239000004205 dimethyl polysiloxane Substances 0.000 description 4
- 238000007689 inspection Methods 0.000 description 4
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 4
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 4
- 239000002699 waste material Substances 0.000 description 4
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 238000004720 dielectrophoresis Methods 0.000 description 3
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- 230000007246 mechanism Effects 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
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- 238000002703 mutagenesis Methods 0.000 description 3
- 231100000350 mutagenesis Toxicity 0.000 description 3
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
-
- G01N15/01—
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N2015/03—Electro-optical investigation of a plurality of particles, the analyser being characterised by the optical arrangement
Abstract
The present invention provides a kind of high flux screening equipment using drop micro-fluidic chip, including：Rack；Microfluidic control system, is configured to control and is used to supply the fluid supply apparatus containing fluids in drops to the drop micro-fluidic chip；Photodetector system, has and limits the incident light generating means of input path, preferably laser excitation device and limits the fluorescence detection device of fluorescence light path；Electric screening system, is configured to connection drop micro-fluidic chip and flows through the drop of the drop micro-fluidic chip to screen；And one or more controllers, it is configured to receive the signal from the fluorescence detection device, and the signal is compared with reservation threshold and is based on the comparative result control electric screening system and carries out drop screening.
Description
Technical field
The present invention relates to the high flux screening equipment using drop micro-fluidic chip, especially unicellular screening installation.This
Invention further relates to the drop screening technique using the equipment.
Background technology
It is always the important way for developing and innovating Research for Industrial Microbial Germ to screen from nature and evolve in laboratory mutagenesis
Footpath, is the basis of industrial biotechnology industry.The microbial strains industrially used now are mainly obtained by mutagenesis screening
's.The screening of microbial strains at present uses Microdilution plate method (96 orifice plates or 384 orifice plates) or flat band method more, these methods are almost
All rely on manual operations, longer time needed to more than 100,000 plants mutation libraries of screening, and need to consume substantial amounts of consumptive material with
Reagent, cost are difficult to bear.Microorganism high-pass Screening Platform utilizes advanced modern automation technology and instrument analysis technology
Realize classic mutagenesis screening process, there is the features such as automation, standardization, high pass quantization, breach artificial screening significantly in speed
The limitation of degree, efficiency and standardization etc..But it is mainly state granddad that all kinds of high flux screening equipment used, which have been put into,
Expensive, the complicated Large-Scale Equipment system of exploitation is taken charge of, in particular for being critical piece with complete using flow cytometer
Into high-throughout requirement, such as the stream type cell analyzer that U.S. company BD provides, its is expensive, and needs special messenger to operate,
Being screened object needs surface or inside to have fluorescence signal, and the metabolite that can not be secreted to cell be detected, screen, and limit
Its application range is made.
Chinese invention patent《A kind of drop style product fluorescence detecting system and method》(Authorization Notice No.
CN104388307A a kind of drop style product fluorescence detecting system and method) are disclosed, still suffers from problems with：(1) liquid is lacked
Drip screening unit；(2) level of integrated system has much room for improvement, and is unfavorable for popularizing.
The Britain's periodical published in 2013《Nature protocols》Disclose document《Single-cell analysis
and sorting using droplet-based microfluidics》(single cell analysis and utilization Microfluidic droplet point
Choosing), it is still in the laboratory research stage, and there is no integrated system scheme.
The Chinese invention patent application of the applicant《A kind of microorganism high-pass detection system based on drop micro-fluidic chip
System》(publication number CN104007091A) discloses a kind of microorganism high-pass detecting system based on drop micro-fluidic chip, but
It is to equally exist similar problems.
The Chinese invention patent application of the applicant《A kind of microscope integrated optical circuit system for drop fluoroscopic examination》
(publication number CN105300943A) provides a kind of light path system for drop fluoroscopic examination, but also without proposition integrated level
High overall drop screening solution.
In this regard, a kind of high high throughput based on drop micro-fluidic chip of and the degree of automation high desirable to provide integrated level sieves
Select solution.It is moreover desirable that scheme has improved drop detection ability.Additionally, it is also desirable to scheme provides improved liquid
Drop feeding.
The content of the invention
For the problem, and the degree of automation high the present invention relates to a kind of integrated level is high based on drop micro-fluidic chip
Unicellular high flux screening equipment, its high throughput fluorescence detection, screening particularly suitable for different type cell.
According to an aspect of the invention, there is provided a kind of high flux screening equipment using drop micro-fluidic chip, bag
Include：Rack；Microfluidic control system, is configured to control and is used to supply the fluid confession containing fluids in drops to the drop micro-fluidic chip
Answer device；Photodetector system, has and limits the incident light generating means of input path and limit the fluorescence of fluorescence light path
Detection device；Electric screening system, the screening electrode for being configured to connection drop micro-fluidic chip flow through the drop miniflow to screen
Control the drop of chip；And one or more controllers, it is configured to receive the signal from the fluorescence detection device, and by institute
Signal is stated compared with reservation threshold and drop screening is carried out based on the comparative result control electric screening system.Incident photogenerated
Device is preferably laser excitation device
Compared to voluntarily building system and the prior art of manpower-intensive type, root according to various concrete conditions in the lab
According to the present invention the high flux screening equipment using drop micro-fluidic chip provide a kind of supermatic drop automatically to
The solution sent, detect and sorted, and can simply be suitable for the detection of various different drops, without building or changing again
Manufacturing apparatus.
In an embodiment according to the present invention, the drop can be in general can fluorescence excitation drop, and in spy
Other preferred embodiment includes the drop of embedding cell, such as embeds single celled drop, and cell for example includes but not limited to big
Enterobacteria, bacillus subtilis or saccharomycete.
In an embodiment of the present invention, screening threshold values various, of different sizes can be set as needed.One
In a preferred embodiment, reservation threshold is, for example, drop fluorescence intensity level.In a preferred embodiment, reservation threshold is big
Small range is whether to be embedded with cell based on drop, be preferably based on whether be embedded with unicellular setting.
According to a preferred embodiment, the fluorescence detection device include one or more imaging lens, baffle of porous baffle and
Photoelectric multiplier.By means of baffle of porous baffle, fluorescence detection device according to embodiments of the present invention can simply and effectively filter out inspection
The fluorescence signal of flanking cell transmitting outside area is surveyed, improves system sorting flux.
According to a particularly preferred embodiment, the hole of the baffle of porous baffle is arranged on the focus upstream in fluorescence light path, excellent
20 μm -100 μm of choosing interval, is more preferably spaced 30 μm -70 μm, is more preferably spaced 50 μm or so.By means of the baffle of porous baffle
It is " preposition " set, it was surprisingly found now that fluorescence detection device according to the preferred embodiment of the invention can ground obtain more preferably
Drop signal detection, the particularly signal detection on drop depth direction.Such as in a preferred embodiment, when drop is equal
During one photosphere, system signal strength detection improves about 32 times.In addition, the fluoroscopic examination with " preposition " baffle of porous baffle fills
The embodiment put also is not affected by significantly affecting for afocal veiling glare.
Preferably, the hole of the baffle of porous baffle is offset to the optical axis setting of the fluorescence light path.The hole of the biasing contributes to
Further improve fluoroscopic examination, such as further reduce the influence of afocal veiling glare.
According to a preferred embodiment, additional convex lens can be also set between baffle of porous baffle and photoelectric multiplier.This
System detectable signal can be further enhanced, and reduces the influence of afocal veiling glare, and the signal for improving photoelectric multiplier receives.
According to a preferred embodiment, the photodetector system is included by the incident light generating means and the fluorescence
Microscope body that detection device shares, be preferably inverted fluorescence microscope main body.Preferably, the microscope body includes using
Exciting light optical filter in input path, transmitting light optical filter, dichroic filter or the first light splitting piece for fluorescence light path
The object lens set with the neighbouring drop micro-fluidic chip.Whereby, the equipment of the preferred embodiment of the present invention can by means of into
The inverted fluorescence microscope main body ripe, integrated level is high, small is transformed, and advantageously reduces manufacture cost, suitable for big rule
Molding is made.
According to a preferred embodiment, the fluorescence detection device includes fluoroscopic examination module, the fluoroscopic examination module
Including shielding case and the baffle of porous baffle and photoelectric multiplier that are contained in shielding case, preferably described fluoroscopic examination module is also
Including the second light splitting piece for being arranged on the baffle of porous baffle upstream being contained in the shielding case and/or the connection photoelectricity times
Increase the digital analog converter of device.Preferably, the shielding case has the first interface and alternatively for being connected to the microscope body
For connecting the second interface of video camera or imaging sensor.Preferably, the first interface is opposite with the second interface
And second light splitting piece is arranged between the first interface and second interface.
The fluoroscopic examination module can be conducive to modularized design, particularly suitable for above-mentioned inverted fluorescence microscope
Transformation, and external interference can be effectively avoided, so as to improve the signal-to-noise ratio of system.
According to a preferred embodiment, the incident light generating means include being connected to the laser of the microscope body
Device.Preferably, the laser is parallel to fluoroscopic examination module, to save space and to be conducive to input path and fluorescence light path
Part coaxial design.
According to a preferred embodiment, the fluid supply apparatus includes one or more syringes, preferably described stream
Body feeding mechanism includes being used to accommodate the first syringe containing fluids in drops and the second syringe for accommodating flow control medium.It is excellent
Selection of land, the microfluidic control system include be used for drive respectively one or more of syringes one or more actuators and
Control the microflow controllers of one or more of actuators.Preferably, the microfluidic control system is configured to miniflow syringe pump.
Compared to the prior art of compression pump form, coordinate and inject the micro-fluidic of the embodiment of the present invention that feeding mechanism uses
System processed, such as injecting pump structure are particularly conducive to being re-injected into for drop, thus provide simple drop supply.This can also be real
Existing drop prepares and supply phase separation, and avoid needs the labyrinth that drop is prepared in situ in the prior art, and also favourable
In saving droplet samples.Moreover, structure according to this embodiment of the invention can also be for example, by being contained in other injection
Flow control medium of the oil as drop in device, so as to realize the accurate control of drop feeding with very simple means.Such as
In one embodiment, equipment according to embodiments of the present invention is configured to exist by one or more controllers or by operating personnel
When fluorescence detection device detects or observes the signal of more than one drop, the flow or flow velocity of flow control medium are improved.
According to a preferred embodiment, the high flux screening equipment is further included for supporting one or more of injections
One or more bearings of device.Preferably, the bearing is configured so that the syringe nozzle upward by vertical or inclination branch
Hold, preferably with 45 ° -90 °, more preferably with 60 ° -90 °, more preferably with 70 ° of -90 ° of inclinations.Preferably, it is one or more of
Bearing also has the fixture for being used for clamping the syringe.It is vertical according to the present invention and be preferably close to vertically tilt supporting
Syringe be particularly conducive to drop it is steady, controllably supply.
According to a preferred embodiment, the drop micro-fluidic chip for the high flux screening equipment has at least
One fluid inlet, two or more fluid outlets, the connection fluid inlet and fluid outlet runner and be connected to
The screening electrode of electric screening system, wherein the runner includes primary flow path and connects the primary flow path and the two or more
Two or more branch flow passages of fluid outlet.Drop micro-fluidic chip for the embodiment of equipment according to the present invention
The similar approach manufacture that the prior art uses can be used, cost can't be increased.The reality for equipment according to the present invention
The drop micro-fluidic chip for applying example is integrated with screening electrode, there is provided simple screening.
Preferably, fluid inlet is configured with vertical inflow segment, this is conducive to the feeding of drop.Preferably, it is described
Primary flow path has narrowing portion at the position being connected with branch flow passage.
Preferably, the screening electrode has cathode and anode.Preferably, screen electrode anode and the end of anode is neighbouring
The connecting portion of the primary flow path and branch flow passage is set.It is highly preferred that by means of the positive and negative anodes of the screening electrode, voltage is screened
Drop application can be adjacent to, such as produces screening electric field, and realizes the reliable screening of drop.
Preferably, the drop micro-fluidic chip is set near the end of the screening electrode anode and anode in upstream
It is useful for the mark of alignment input path.By means of the setting of the mark, input path precisely inspection required for alignment can be realized
The drop of survey, so as to realize the accurate detection of drop signal, and can be precisely controlled and screen drop to be screened.It is excellent
Choosing, the mark can be harmonious with predetermined drop conveying relative to the position of positive and negative anodes end, so as to further improve pair
The accurate control of drop to be screened.
Preferably, the mark and the screening electrode are respectively provided at the both sides of the sprue.
According to a preferred embodiment, the electricity screening system is preferably dielectrophoresis screening system and including high voltage amplifier
Device, so that more reliable screening voltage is provided, so as to for example provide the dielectrophoresis force of abundance.Preferably, the high voltage amplifier
Device includes power switch, the signal input part being connected with one or more of control devices, high-voltage output end and ground terminal.
Preferably, the high-voltage output end and ground terminal are configured to be connected to the screening electrode of drop micro-fluidic chip, it is preferable that described
High-voltage output end is connected to the cathode of the screening electrode of drop micro-fluidic chip, and the ground terminal is connected to drop micro-fluidic chip
Screening electrode anode.
According to a preferred embodiment, one or more of control devices include signal processing and analysis system and control
Case.Preferably, the signal processing and analysis system receives the signal from the fluorescence detection device and by the signal and in advance
Determine threshold values to compare.Preferably, the control cabinet has incident light generating means switch and/or control module, fluoroscopic examination dress
Switch and/or control module are put, and/or electric sieve selects system control module.Preferably, the electric sieve selects system control module
For controlling the screening voltage of electric screening system.These embodiments provide control device that is simple in structure and being easily achieved and implement
Scheme.
In an alternative embodiment, one or more of control devices include master controller, be preferably band PC or
Master controller with display.
According to a preferred embodiment, the rack is two layers of open frame.Preferably, the microfluidic control system is placed on
Upper strata, the photodetector system and electric screening system are placed on lower floor.Preferably, the photodetector system is to be inverted to set
And with the top bearings for being used to support the drop micro-fluidic chip so that the microfluidic control system is higher than described
Top bearings.This embodiment offers highly integrated and compact-sized equipment.
Preferably, which may also include the housing for each building block of enclosing equipment, and the housing can include using
Can pivot in first pivotable cover in covering fluid supply apparatus region and/or second for placing drop micro-fluidic chip region
Hub cap.
According to an aspect of the present invention, drop screening technique is also provided, especially with according to embodiments of the present invention
The drop screening technique of equipment.
According to another aspect of the invention, for example the Photoelectric Detection system for equipment according to embodiments of the present invention is also provided
System, fluorescence detection device and/or fluoroscopic examination module.
In accordance with a further aspect of the present invention, also providing the inverted fluorescence microscope transformed according to the present invention and transformation should fall
The method for putting fluorescence microscope.
Compared with prior art, the present invention has the advantages that：
(1) present invention provide it is a kind of based on drop micro-fluidic chip can fluorescence excitation drop, especially unicellular screening
Equipment, chip flexible design, replace simply, and processing and fabricating cost is low；Modularized design, is conveniently adjusted, and detection range is wider；From
Dynamicization controls, and saves human cost；On-line checking, saves the time；Easy to operate, cost is low, convenient to promote.Thus, it is of the invention
Equipment there is very high the degree of automation and integrated level, and the equipment can be utilized to can be applied to the height of different type cell
Throughput fluorescence detects and screening, without setting for this platform again.
(2) equipment of the preferred embodiment of the present invention can improve the detection result to drop, especially by novelty
The structure design of electro-optical system, can obtain more droplet samples information, the especially information of depth direction.
(3) equipment of the preferred embodiment of the present invention can use improved drop feeding scheme, not only be particularly conducive to liquid
Drop is re-injected into, and also helps the accurate control of drop feeding.
(4) equipment of the preferred embodiment of the present invention can be by means of the inversion fluorescence ripe, integrated level is high, small
Microscope body is transformed, and advantageously reduces manufacture cost, suitable for extensive manufacture.
A part for other features and advantages of the present invention would is that those skilled in the art are obvious after the application is read
, another part will be described in conjunction with the accompanying in embodiment below.
Brief description of the drawings
Hereinafter, the embodiment that the present invention will be described in detail is carried out with reference to attached drawing, wherein：
Fig. 1 shows the drop micro-fluidic based on liquid of embodiment of the present invention, especially unicellular screening installation
Schematic diagram.
Fig. 2 shows the drop micro-fluidic based on liquid of embodiment of the present invention, especially unicellular screening installation
Operation state views.
Fig. 3 shows the drop micro-fluidic based on liquid of embodiment of the present invention, especially unicellular screening installation
Off working state view.
Fig. 4 shows the schematic diagram of the microfluidic control system of embodiment of the present invention.
Fig. 5 shows the structure chart of the microfluidic control system of embodiment of the present invention.
Fig. 6 shows an embodiment of the drop micro-fluidic chip of the screening installation for embodiment of the present invention.
Fig. 7 shows the schematic diagram of the photodetector system of the screening installation of embodiment of the present invention.
Fig. 8 shows the light path schematic diagram of fluoroscopic examination module according to embodiments of the present invention.
Fig. 9 shows the schematic diagram of the signal analysis and processing software module being stored in PC of embodiment of the present invention.
Figure 10 shows the schematic diagram of the display interface of the signal analysis and processing software of embodiment of the present invention, shows inspection
The drop signal and screening threshold values measured.
Figure 11 diagrammatically illustrates the drop micro-fluidic based on liquid according to embodiments of the present invention, especially unicellular
Screening installation, wherein shell are removed for clearly showing internal structure.
Figure 12 diagrammatically illustrates the high-level schematic functional block diagram of the screening installation of another embodiment according to the present invention.
Reference numerals list
1st, 1 '-microfluidic control system；
11- microflow controllers；12- syringes；13- actuators；14- bearings；141- pedestals；142- fixtures；
2nd, 2 '-drop micro-fluidic chip；
21-PDMS layers；22- glassy layers；23- fluid inlets；24- fluid outlets；25- screens electrode anode；26- screening electricity
Pole anode；27- runners；271- primary flow paths；272nd, 272 '-branch flow passage；28- narrowing portions；29- is marked；
3rd, 3 '-photodetector system；
31- laser excitation modules；31 '-laser excitation device；310th, 310 '-input path；311st, 311 '-laser；
312 '-optics；
32- fluoroscopic examination modules；32 '-fluorescence detection device；320th, 320 '-fluorescence light path；321- spectroscopes；322- bands
Hole baffle；3220- holes；323- photomultipliers；323 '-detector；324- high-speed cameras；325- shielding cases；3251- first
Interface；3252- second interfaces；329- digital analog converters；
33- control cabinets；331- total power switch；332- laser controls switch；333- photomultiplier controlling switches；
334- screens voltage control module；
34- inverted fluorescence microscope main bodys；343- exciting light optical filters；344- dichroscope optical filters；345- launches light
Optical filter；346- object lens；
37- objective tables；
4- signal processing and analysis systems；4 '-master controller；
41、41’-PC；42- signal analysis and processing softwares；421- screening threshold value settings；422- trigger delays are set；423-
Trigger duration setting；424- gain of photomultipliers are set；The drop signal that 425- is detected；426- screens threshold value
5- electricity screening systems；
51- high-voltage amplifiers；511- power switches；512- signal input parts；513- high-voltage output ends；514- ground terminals；
6- racks；
7- shells；The first pivotable covers of 71-；The second pivotable covers of 72-；73- button fly fronts；74- button fly fronts；
8- containers；81- drop receptacles；82- waste fluid containers.
Embodiment
Although it is in order to which some embodiments of the present invention are presented to provide attached drawing, attached drawing need not be by specific embodiment
Size is drawn, and some features can be exaggerated, remove or cutting is preferably to show and explain the disclosure.It is attached
Partial component in figure can carry out position adjustment according to the actual requirements on the premise of not influence technique effect.Go out in the description
Existing phrase " in the accompanying drawings " or similar term need not refer to all attached drawings or example.
Here, directional word (for example, top, lower section, it is upward, downward, left, right, to
It is left, to the right, top, bottom, above, following, vertically, horizontal, clockwise and counterclockwise) only use
It is of the invention to help reader to understand in the purpose of identification, and it is unrestricted, particularly to the position of the present invention, direction or make
Limitation.
In addition, term " first ", " second " and its similar terms used in the present invention, are not offered as in the present invention
Any order, quantity or importance, but for a component to be distinguished with other components.
With reference to the high throughput using drop micro-fluidic chip 2 for reference to figure 1-3, showing embodiment according to the present invention
Screening installation.The especially single celled high flux screening equipment of the high flux screening equipment.Although it will be directed to below unicellular
High flux screening embodiment of the present invention is described, it will be obvious to those skilled in the art that according to the present invention high throughput sieve
Optional equipment can be also used for a variety of drops, such as any suitable drop for exciting fluorescence.
Fig. 1 shows the structural representation of the functional subsystem of the high flux screening equipment of embodiment according to the present invention
Figure.The embodiment according to the present invention, may include to be used to control stream using the high flux screening equipment of drop micro-fluidic chip 2
Microfluidic control system 1, photodetector system 3, signal processing and analysis system 4 and the electric screening system 5 of body feeding mechanism.In addition,
With reference to shown in Fig. 2-3 and Figure 11, which may also include the rack 6 and shell 7 for supporting each functional component.
The function module and related accessory of the high flux screening equipment of the embodiment are described respectively below with reference to the accompanying drawings.
With reference to the microfluidic control system 1 for reference to figure 1-3 and Fig. 4-5, describing embodiment according to the present invention.Shown
In embodiment, which can be configured to or including miniflow syringe pump.The microfluidic control system 1 or miniflow injection
Pump may include (syringe pump) microflow controllers 11 and actuator 13.In the embodiment illustrated, which can use
As one or more syringes 12 of fluid supply apparatus or one or more syringes are equipped with control.In Fig. 4 institutes
In the embodiment shown, which has used two syringes 12, wherein the first syringe 121 accommodates fluids in drops (such as (fluorine
Change) Water-In-Oil formula drop), drop be preferably embedding cell drop, the second syringe 122 is accommodated for controlling fluids in drops to exist
The flow control medium of conveying in micro-fluidic chip, such as (be fluorinated) oil.In some embodiments of the present invention, made using syringe
High flux screening equipment for fluid supply apparatus can arrange or not arrange general or special purpose syringe, and may be selected not
Same syringe number, this is within the scope of the present invention.
In use, microflow controllers 11 can control the action of actuator 13, so as to control syringe 12 to feed fluid
Speed.In a preferred configuration, microflow controllers 11 can be used for the stream for the syringe 122 for accommodating flow control medium by adjusting
Amount, and adjustment control drop is by the speed of micro-fluidic chip 2, thus be conducive to the detected downstream to drop, screening it is accurate
Control.
With reference to shown in Fig. 2-5, which can include being used for the one or more bearings 14 for supporting syringe, such as 3
Bearing 14.As shown in figure 11, bearing 14 can be supported in 6 upper strata of rack.Bearing 14 can have the pedestal 141 and folder being obliquely installed
Tool 142, is supported in bearing so that syringe 12 can be tilted upward with the head of a quilt.Head-up it is obliquely installed, especially by this
It is head-up obliquely installed close to vertical angles (such as from about 70 °), can high quality, feed drop higher degree of controllability.
With reference to reference to figure 1 and Fig. 6, show that the drop for high flux screening equipment according to embodiments of the present invention is micro-
Fluidic chip 2.In some embodiments of the present invention, can be arranged using the high flux screening equipment of drop micro-fluidic chip
Or general or special purpose drop micro-fluidic chip is not arranged, this is within the scope of the present invention.
In the embodiment depicted in fig. 6, drop micro-fluidic chip 2 may include in the PDMS layers 21 on upper strata and in lower floor
Glassy layer 22.Preferably, the PDMS layer 21 and glassy layer 22 can be bonded by plasma, form the micro-fluidic core of drop
Piece.In shown preferred embodiment, PDMS layers 21 can etch fluid passage by soft lithography, but can also use it
Its fluid channel forming method.
With continued reference to Fig. 6, in the embodiment illustrated, the drop micro-fluidic chip 2 includes two fluid inlets
231st, 232, two fluid outlets 241,242, the runner 27 of the connection fluid inlet 23 and fluid outlet 24 and be connected to
The screening electrode of electric screening system.In embodiment as shown in Figure 6, the runner 27 is included described in primary flow path 271 and connection
Two branch flow passages 272,272 ' of primary flow path and described two fluid outlets.In the embodiment illustrated, the primary flow path
271 have narrowing portion 28 at the position being connected with branch flow passage 272,272 '.
In embodiment as shown in Figure 6, such as the first syringe 121 for accommodating fluids in drops can be configured to
First fluid entrance 231 is connected by micro-pipe, such as hose, the second syringe 122 for accommodating flow control medium can be configured to
Second fluid entrance 232 is connected by micro-pipe, such as hose.The fluid outlet of drop micro-fluidic chip 2 can respectively by micro-pipe,
As hose is connected with container 8, such as centrifuge tube.Such as in the embodiment illustrated, first fluid outlet 241 can be connected by micro-pipe
Screening drop receptacle 81 is connected to, second fluid outlet 242 can be connected to waste fluid container 82 by micro-pipe.
With continued reference to Fig. 6, which preferably has cathode 25 and anode 26.Screen electrode anode 25 and anode 26
End can be arranged on the first side of primary flow path 271 adjacent to the connecting portion of the primary flow path and branch flow passage.Opposite second
Side, the chip 2 can also be equipped with the mark 29 for being used for being directed at input path.The end of the neighbouring screening electrode of the mark 29, but set
In upstream.
Although show that drop is re-injected into an embodiment of screening chip 2, ability in the embodiment illustrated
Field technique personnel are readily apparent that equipment according to the present invention can be re-injected into screening chip or other with reference to the drop of other forms
The chip that screening chip, such as drop formation screening chip used or combined other functions uses.In shown embodiment
In, the drop micro-fluidic chip that high flux screening equipment uses is replaceable, and which greatly enhances suitability.Unshowned
In embodiment, culture whether can be needed to select drop micro-fluidic chip according to chip functions and corresponding detectable substance.
With reference to reference to figure 1-3 and Fig. 7-8, photodetector system 3 according to embodiments of the present invention is described.In the embodiment party
In case, photodetector system 3 includes limiting the incident light generating means of input path 310, and the incident light generating means are preferred
For laser excitation device.Photodetector system 3 may also include the fluorescence detection device for limiting fluorescence light path 320.
In the embodiment shown in Fig. 1-3 and Fig. 7-8, the photodetector system 3 can include fluorescence microscope main body,
Particularly inverted fluorescence microscope main body 34, or can be obtained by transforming (inversion) fluorescence microscope.
With specific reference to Fig. 1 and Fig. 7, the incident light generating means and fluorescence detection device of the photodetector system 3 can be total to
With (inversion) the fluorescence microscope main body 34, so that preferably input path 310 and 320 part of fluorescence light path is coaxial.Reference chart
1 and Fig. 7, the incident light generating means (laser excitation device) and fluorescence detection device can include being connected to (inversion) respectively
The laser excitation module 31 and fluoroscopic examination module 32 of fluorescence microscope main body 34.The shared fluorescence microscope main body is favourable
Cost is manufactured in reducing, and can be by providing for simple replacement of or adjusting relevant excitation module to the Adjustment and reform of equipment light path system
And/or detection module is realized, commutativity is high and suitability is good.
With reference to the embodiment shown in figure 1 and Fig. 7, being somebody's turn to do (inversion) fluorescence microscope main body 34 may include that dichroscope filters
Piece 344 (or first light splitting piece), thus coaxial input path 310 and fluorescence light path 320 preferred setting at 45 ° two to
Look mirror optical filter 344 (or first light splitting piece) place is separated.Preferably, being somebody's turn to do (inversion) fluorescence microscope main body 34 may include to use
Exciting light optical filter 343 in input path 310, the transmitting light optical filter 345 for fluorescence light path.(inversion) fluorescence microscopy
Exciting light optical filter 343, dichroscope optical filter 344 and the transmitting light optical filter 345 of mirror main body 34 can be according to detection materials
Difference replaces the optical filter of different wave length.
It is somebody's turn to do the object lens 346 that (inversion) fluorescence microscope main body 34 may also include adjacent chips setting.
In the embodiment illustrated, which can also utilize the loading that (inversion) fluorescence microscope (main body) arranges
Supporting part of the platform 37 as chip placement, but it is contemplated that other types of supporting part.
With continued reference to Fig. 1 and Fig. 7, which can be or including laser 311.In shown embodiment party
In case, which is connected to the expansion interface of 34 side of inverted fluorescence microscope main body and is preferably parallel to glimmering
Light detection module 32 is set, so that the fluorescence that incident light and fluoroscopic examination module 32 that for example laser excitation module 31 is sent receive
Almost parallel (and being each perpendicular to coaxial light path part).
With reference to reference to figure 1 and Fig. 7-8, fluorescence detection device according to embodiments of the present invention or fluoroscopic examination module are described
32.Fluoroscopic examination module 32 may include for example cylindrical circular shielding case 325, the second light splitting piece being contained in shielding case 325
321st, baffle of porous baffle 322 and photoelectric multiplier, such as photomultiplier 323.In shown preferred embodiment, the fluorescence inspection
Digital analog converter 329 can also be included by surveying module 32, and the digital analog converter 329 can pass through data cable and the photomultiplier
323 are connected.But the digital analog converter can also be arranged in signal processing and analysis system 4 in other embodiments.It is described
Second light splitting piece 321, baffle of porous baffle 322, photomultiplier 323 can be totally-enclosed by shielding case 325, and shielding case 325 is preferred
For ferrous metal shell.
Second light splitting piece 321 can be placed in before baffle of porous baffle 322 and preferably setting at 45 °, and the plate washer 322 with holes can
It is placed on before photomultiplier 323.The shielding case 325 has the first interface 3251 for being connected to microscope body and is used for
Connect video camera or imaging sensor, the second interface 3252 of such as high-speed camera 324.The first interface 3251 and described the
Two interfaces, 3252 opposite and described second light splitting piece 321 is arranged between the first interface 3251 and second interface 3252.
In shown preferred embodiment, second light splitting piece 321 arrives baffle of porous baffle 322 and the distance to high-speed camera 324
It is equal.
Focus (not shown), this example are formed in the fluorescence light path of fluorescence detection device in the embodiment in accordance with the present invention
Being formed in this way by one or more imaging lens (such as imaging convex lens in object lens in the present embodiment).In Fig. 8
In shown preferred embodiment, the hole 3220 of the baffle of porous baffle 322 is arranged on the focus upstream in fluorescence light path and preferably
The optical axis for being offset to fluorescence light path is set.In a preferred embodiment, hole 3220 relative to focus " preposition " at intervals of
About 20 μm -100 μm, even more preferably about 30 μm -70 μm, more preferably 50 μm or so.In shown preferred embodiment,
Additional convex lens 326 can be also set between baffle of porous baffle 322 and photomultiplier 323.It is preferred that about 20 μm -100 μm of biasing,
Even more preferably about 30 μm -70 μm, more preferably 50 μm or so.Although being not bound by the limitation discussed, the hole for being somebody's turn to do " preposition " is particularly advantageous
In the more drop fluorescence informations of acquisition particularly in the depth direction；And the hole of biasing can be conducive to avoid adjacent drops
Effect of signals；The detection that above-mentioned additional convex lens is equally beneficial for drop information obtains.
It may include or be connected to control cabinet 33 with reference to reference to figure 1, Fig. 7 and Figure 11, photodetector system 3.The control cabinet 33
May include total power switch 331, laser switch 332 or control module, photomultiplier switch 333 or control module and
Electric sieve selects system control module, such as screens voltage control module 334.Correspondingly, the control cabinet 33 can be distinguished by data cable
It is connected with the laser 311, photomultiplier 323.In addition, the control cabinet 33, which is additionally coupled to electric sieve, selects control system 5.
With reference to the signal processing and analysis system 4 for reference to figure 1 and Fig. 9-10, showing embodiment according to the present invention.
In shown embodiment, signal processing and analysis system 4 includes the PC41 with signal analysis and processing software 42, but is contemplated that
The signal processing and analysis system 4 of embodiment according to the present invention may include any to gather fluorescence signal and carry out corresponding
Analysis and the controller and/or signal analysis and processing module or program of control.In the embodiment illustrated, signal acquisition point
Analysis system 4 is connected to the digital analog converter 329 of fluoroscopic examination module 32, but as previously described, it is contemplated that in signal processing and analysis
Integrated digital to analog converter in system, and the digital analog converter of signal processing and analysis system may be connected directly to fluorescence detection device/
The photoelectric multiplier of module, such as photomultiplier.
The signal analysis and processing software 42 may include various program modules, such as screen threshold value setting 421, trigger delay is set
Fixed 422, triggering duration setting 423, gain of photomultiplier setting 424, the drop signal value 425 detected and screening threshold
Value 426.The drop signal value 425 and screening threshold value 426 detected as described in showing by display interface Figure 10.But this area
Technical staff is it will be appreciated that some or all of above procedure module can pass through hardware realization.Sampling and analysis of signal system
System 4 is configured to receive the signal from the fluorescence detection device or photomultiplier and by the signal and predetermined screening
Threshold values compares, and when detecting drop signal value more than screening threshold values, signals to control cabinet 33 to control electric screening system 5
Screening is performed, such as causes corresponding drop to flow into drop receptacle 81 by branch flow passage 272.And the drop signal for detecting
Not less than the drop of screening threshold values, then it is allowed to flow into (for example electric screening system 5 at this time of waste fluid container 82 through branch flow passage 272 '
Electric field is not applied to the drop).In the different embodiments of the present invention, screening threshold values can be one or more, such as have
Have and screen threshold values as the first of lower limit and screen threshold values as the second of the upper limit, for example to fall not for screening (discarded) respectively
The drop of embedding cell and the drop for embedding multiple cells.
Furthermore it is preferred that based on mark and screening electrode tip spacing, can by set trigger delay and delay when
Between with detecting exceed predetermined value when, in an optimal manner apply electric field to intend screening drop.
With continued reference to Fig. 1, electricity screening system 5 according to embodiments of the present invention can be dielectrophoresis screening system, excellent
Choosing includes high-voltage amplifier 51.In shown preferred embodiment, the high-voltage amplifier 51 may include power switch 511,
Connect signal input part 512, high-voltage output end 513 and the ground terminal 514 of control cabinet 33.High-voltage output end 513, ground terminal 514
It is connected by cathode 25, anode 26 of the power cord respectively with the screening electrode channel of drop micro-fluidic chip 2.
With reference to figure 11, which is double-deck rack.In micro-injection pump form microfluidic control system 1 (and injection
Device bearing) it can be placed in upper strata.Photodetector system 3 and electric screening system 5 are placed on lower floor.Photodetector system 3 is to be inverted
Formula, and upper strata is extended to, so that micro-injection pump 11 is slightly above inverted objective table 37.The inversion fluorescence of photodetector system 3
Microscope body 34 is arranged in the middle part of lower floor, and electric screening system is arranged on the left side of inverted fluorescence microscope main body 34, the control
Case 33 is positioned over the right side of inverted fluorescence microscope main body 34.
Referring to figs. 2 and 3 multiple building blocks of the equipment are surrounded by shell 7, and cover top portion is provided with first and
Two pivotable covers 71,72 are to be covered each by 2 region of bearing 14 and chip.Shell 7 is also provided with two button fly fronts 73,74 to hide
Keep off control cabinet 33 and high-voltage amplifier 51.
The drop micro-fluidic based on drop of an embodiment according to the present invention, especially unicellular screening installation
The course of work may include following steps：
(1) drop micro-fluidic chip 2 is fixed on objective table 37, by the first syringe 121 containing drop and contained
Second syringe 122 of flow control medium such as fluorinated oil passes through micro-pipe the first and second fluids with drop micro-fluidic chip 2 respectively
Entrance 231,232 is connected, and the first and second fluid outlets 241,242 are connected by micro-pipe with container 8, such as centrifuge tube respectively, high
Press amplifier 51 by power cord respectively by high-voltage output end 513, the screening electrode of ground terminal 514 and drop micro-fluidic chip 2
Cathode 25, the anode 26 of passage are connected；
(2) alternatively, it is most clear that drop micro-fluidic chip 2 is presented in the position of adjustment objective table 37 on high-speed camera 324
The picture of Chu；
(3) micro-injection pump 11 is opened, alternatively suitably adjustment fluorinated oil, drop flow velocity, drop all flow to second point
Zhi Liulu (waste fluid container)；
(4) laser control switch 332 is opened, alternatively adjusts laser spot position into 2 passage of drop micro-fluidic chip
Centre；
(5) photomultiplier controlling switch 333, the power switch 511 of high-voltage amplifier 51, opening signal collection point are opened
The signal processing and analysis software 42 of analysis system 4；It alternatively can perform following one or more sub-steps：Setting screening threshold value setting
421st, trigger delay setting 422, triggering duration setting 423, gain of photomultiplier setting 424；
(6) drop signal processing and analysis and drop screening are carried out.
With reference to figure 12, the screening using drop micro-fluidic chip 2 ' of another embodiment according to the present invention is shown
The high-level schematic functional block diagram of equipment.In this embodiment, screening installation may include the microfluidic control system 1 ' based on syringe,
Photodetector system, is equipped with the master controller 4 ' of PC41 ' and electric screening system 5 '.The photodetector system may include to limit
The incident light generating means of input path 310 ', preferably laser excitation device 31 ' and the fluorescence for limiting fluorescence light path 320 '
Detection device 32 '.In the embodiment illustrated, microfluidic control system 1 ' is equipped with 4 syringes 12 ', two of which injection
Device for example accommodates fluids in drops, two syringes accommodate flow control medium.Correspondingly, drop micro-fluidic chip 2 ' can include four
Fluid inlet.Preferably, fluid inlet can have vertical inflow segment.In shown chip 2 ', fluids in drops entrance is connected
Runner preferably there are multiple meanders.
Difference lies in the implementation shown in Figure 12 by one of the embodiment shown in embodiment and Fig. 1 shown in Figure 12
Incident light generating means 31 ' and the unshared component of fluorescence detection device 32 ' in scheme.In the embodiment depicted in fig. 12, it is glimmering
Optical detection device 32 ' includes one or more imaging lens 321 ' and for receiving the detector 323 ' of fluorescence information, such as photoelectricity
Multiplier and/or video camera or imaging sensor.Although not shown in embodiment in fig. 12, it is contemplated that Fig. 1-11
Shown in embodiment in the feature of photodetector system the embodiment party of Figure 12 can be bound in the case of reconcilable
Case.Particularly, baffle of porous baffle can be preferably set in the fluorescence detection device 32 ' in the embodiment of Figure 12, it for example sets
Put before detector, such as photoelectric multiplier.The hole of baffle of porous baffle similarly preposition in focus and/or can be offset to fluorescence light path
Optical axis.In the embodiment depicted in fig. 12, laser excitation device 31 ' may include laser 311 ' and one or more optics
Device 312 '.
In embodiment shown in Figure 12, master controller 4 ' is configured to receive the letter from the fluorescence detection device 32 '
Number, and the signal is compared with reservation threshold and based on the liquid of comparative result control electric the making choice property of screening system
Drop screening.Thus, embodiment shown in Figure 12 and Fig. 1 another difference is that, lead in the embodiment depicted in fig. 12
Cross the master controller with PC and realize various control function, it is such as, but not limited to above-mentioned by control cabinet 33 and signal processing and analysis system
The function of system 4.But it may occur to persons skilled in the art that the one or more controllers taken other form are (as individually
With or without PC or the master controller with other accessories such as display) receive detection signal to realize, analysis and trigger
The function of screening, and other functions are also alternatively integrated, such as microfluidic control function, this is within the scope of the present invention.
Embodiment：Express the bacillus subtilis drop fluoroscopic examination of red fluorescent protein
LB liquid is dropped down onto from picking single bacterium on the bacillus subtilis LB solid plates of fresh expression red fluorescent protein
Culture medium, 37 DEG C, 200rpm cultivate between OD600=0.6-0.8, with the PBS buffer washing thalline 3 times of pH7.4, often
Secondary 775g, 5min, are finally resuspended thalline with PBS buffer, by mycelium dilution to OD600=0.125, select cell embedding chip
The drop embedding of cell is carried out, sets 200 μ L/h of Oil phase flow rate, B. subtilis cell dilution flow velocity is 50 μ L/h, raw
In into drop to 1mL syringes, liquid-drop diameter is 20 μm, is 0.6065 without cell drop ratio according to Poisson distribution, embedding
Unicellular drop ratio is 0.3033, and embedding two and above cell drop ratio are 0.0902.
Drop detection chip 2 is positioned on objective table 37,37 position of adjustment objective table makes drop detection chip 2 at a high speed
Blur-free imaging on video camera.Drop flow velocity is set as 10 μ L/h, Oil phase flow rate is 200 μ L/h, and drop is injected drop detection core
Piece 2, opens laser 311, and 2 position of adjustment drop detection chip makes the Air conduct measurement point of laser alignment drop detection chip 2.
Open photomultiplier 323, the gain of the activation threshold value, photomultiplier of setting signal interpretation software 42
Value, oscillogram show activation threshold value and drop fluorescence intensity level, as shown in fig. 6, horizontal line is to screen threshold marker line, vertical line in figure
For drop fluorescent value.Thus, when the detected value of detected object droplet exceedes predetermined screening threshold values, electric sieve is automatically controlled
System 5 is selected to screen the object droplet for flowing through chip 2.
Illustrate that " one " means " one or more " for disclosed purpose and unless there are it.Just in this specification
For the term " comprising " used in claims or " including ", it will be inclusively, this class to a certain extent
"comprising" is similar to, because those terms are explanatory when as transition conjunction.In addition, for term "or" used
(such as A or B), it will imply that " A or B or both ".When applicant intends to show " only A or B but non-both ", it will
Use " only A or B but non-both ".Therefore, the use of term "or" be include and it is non-exclusive.Referring to
Bryan.A.Garner's《Modern Law dictionary》Page 624 (2d.Ed.1995).Also with regard to specification or claims
Term " in " used or for " inside ", they are also intended to the other meaning " upper "
Or " above ".In addition, for term " connection " used in specification or claims, it will not only anticipate
Taste " being directly connected to ", also means " being connected indirectly to ", is such as connected by another component or multiple components.Such as this paper institutes
" about " it will be understood by those skilled in the art and can be changed to a certain extent according to occasion used in it.
If the use of the term is unclear to those skilled in the art, then " about " be will imply that to particular term extremely
It is add deduct 10% more.It will imply that, from about X to about Y, X, Y are occurrences herein from about X to Y.
With reference to the exemplary system and method that the embodiment above specifically illustrates and the invention has been described, it is only to implement
The example of the optimal mode of the system and method.It will be appreciated by those skilled in the art that be can implement the system and/
Or various changes are done during method to the embodiment of system and method described herein without departing from being defined in appended claims
In spirit and scope of the present invention.Appended claims are intended to define the scope of the system and method, therefore fall into these rights
It can be included in it is required that and with the system and method that it is equal.This should be read to include to the above description of the system and method
In the combination of whole new and non-obvious elements that describes, and may be present in the application or subsequent application be related to it is any
The claim of the combination of new and non-obvious element.In addition, the embodiment above is exemplary, in this Shen
Please or subsequent application in can be claimed may all combine, neither one single features or element be essential
's.
Claims (10)
1. a kind of high flux screening equipment using drop micro-fluidic chip, it is characterized in that, including：
Rack；
Microfluidic control system, is configured to control and is used to supply the fluid supply dress containing fluids in drops to the drop micro-fluidic chip
Put；
Photodetector system, has and limits the incident light generating means of input path, preferably laser excitation device and restriction
Go out the fluorescence detection device of fluorescence light path；
Electric screening system, is configured to connection drop micro-fluidic chip and flows through the drop of the drop micro-fluidic chip to screen；With
And
One or more controllers, are configured to receive the signal from the fluorescence detection device, and by the signal with making a reservation for
Threshold values, which compares and is based on the comparative result control electric screening system, carries out drop screening.
2. high flux screening equipment according to claim 1, it is characterized in that, the fluorescence detection device includes one or more
A imaging len, baffle of porous baffle and photoelectric multiplier；Preferably, the hole of the baffle of porous baffle is arranged on the focus in fluorescence light path
Upstream, preferably by 20 μm -100 μm, is more preferably spaced 30 μm -70 μm, is more preferably spaced 50 μm or so；Preferably, the band
The optical axis that the hole of hole baffle is offset to the fluorescence light path is set；Preferably, also set between baffle of porous baffle and photoelectric multiplier
It is equipped with additional convex lens.
3. high flux screening equipment according to claim 2, it is characterized in that, the photodetector system include by it is described enter
Penetrate microscope body that photogenerated device and the fluorescence detection device share, be preferably inverted fluorescence microscope main body, preferably
The ground microscope body includes the exciting light optical filter for input path, the transmitting light optical filter for fluorescence light path, two
The object lens set to colo(u)r filter or the first light splitting piece and the neighbouring drop micro-fluidic chip.
4. high flux screening equipment according to claim 3, it is characterized in that,
The fluorescence detection device includes fluoroscopic examination module, and the fluoroscopic examination module includes shielding case and is contained in shielding case
In the baffle of porous baffle and photoelectric multiplier, preferably described fluoroscopic examination module, which further includes, to be contained in the shielding case
The second light splitting piece of the baffle of porous baffle upstream and/or the digital analog converter of the connection photoelectric multiplier are arranged on, preferably
The shielding case have the first interface that is connected to the microscope body and for connecting video camera or imaging sensor the
Two interfaces, preferably described first interface is opposite with the second interface and second light splitting piece is arranged on described first and connects
Between mouth and second interface；And/or
The incident light generating means include being connected to the laser of the microscope body, preferably described laser parallel to
Fluoroscopic examination module.
5. the high flux screening equipment according to one of Claims 1-4, it is characterized in that, the fluid supply apparatus includes
One or more syringes, preferably include the first syringe for accommodating fluids in drops and second for accommodating flow control medium
Syringe；The microfluidic control system includes being used for the one or more actuators for driving one or more of syringes respectively
With the microflow controllers for controlling one or more of actuators；Preferably, the microfluidic control system is configured to miniflow injection
Pump.
6. high flux screening equipment according to claim 5, it is characterized in that, the high flux screening equipment, which further includes, to be used for
Support one or more bearings of one or more of syringes；Preferably, the bearing is configured so that the syringe
Head-up supported by vertical or inclination, preferably with 45 ° -90 °, more preferably with 60 ° -90 °, more preferably with 70 ° of -90 ° of inclinations；
Preferably, one or more of bearings also have the fixture for being used for clamping the syringe.
7. the high flux screening equipment according to one of Claims 1-4, it is characterized in that, set for the high flux screening
The standby drop micro-fluidic chip has at least one fluid inlet, two or more fluid outlets, the connection fluid
The runner of entrance and fluid outlet and the screening electrode for being connected to electric screening system, wherein the runner includes primary flow path and company
Connect two or more branch flow passages of the primary flow path and the two or more fluid outlets；Preferably, the screening
Electrode has cathode and anode, it is preferable that the end of screening electrode anode and anode is adjacent to the primary flow path and branch flow passage
Connecting portion is set；Preferably, the drop micro-fluidic chip it is described screening electrode anode and anode end near upper
Trip is provided with the mark for being directed at input path, it is preferable that the mark and the screening electrode are respectively provided at the mainstream
The both sides in road.
8. the high flux screening equipment according to one of Claims 1-4, it is characterized in that, the electricity screening system includes height
Press amplifier；Preferably, the letter that the high-voltage amplifier includes power switch, is connected with one or more of control devices
Number input terminal, high-voltage output end and ground terminal；Wherein, the high-voltage output end and ground terminal are configured to be connected to drop micro-fluidic
The screening electrode of chip, it is preferable that the high-voltage output end is connected to the cathode of the screening electrode of drop micro-fluidic chip, described
Ground terminal is connected to the anode of the screening electrode of drop micro-fluidic chip.
9. the high flux screening equipment according to one of Claims 1-4, it is characterized in that, one or more of control dresses
Put and received including signal processing and analysis system and control cabinet, the signal processing and analysis system from the fluorescence detection device
Signal simultaneously compares the signal with reservation threshold, it is preferable that the control cabinet have incident light generating means switch and/or
Control module, fluorescence detection device switch and/or control module, and/or electric sieve select system control module, it is preferable that institute
Stating electric sieve selects system control module to be used for the screening voltage for controlling electric screening system.
10. the high flux screening equipment according to one of Claims 1-4, it is characterized in that, the rack is two-layer machine
Frame, wherein, the microfluidic control system is placed on upper strata, and the photodetector system and electric screening system are placed on lower floor, excellent
Photodetector system described in selection of land is inversion type and limits the top bearings for supporting the drop micro-fluidic chip
So that the microfluidic control system is higher than the top bearings.
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| CN108844936A (en) * | 2018-07-09 | 2018-11-20 | 领航基因科技（杭州）有限公司 | A kind of three-dimensional drop detection system and detection method |
| CN109735429A (en) * | 2019-01-28 | 2019-05-10 | 佛山市铬维科技有限公司 | Micro-fluidic chip and the system and its separation method for separating various kinds of cell |
| WO2020011193A1 (en) * | 2018-07-11 | 2020-01-16 | The University Of Hong Kong | Automatic microfluidic system for continuous and quantitive collection of droplets |
| WO2020063864A1 (en) * | 2018-09-29 | 2020-04-02 | 青岛华大智造普惠科技有限公司 | Microfluidic chip system and method for preparing liquid drop |
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