CN110116028A - Microfluidic experimental device and method - Google Patents
Microfluidic experimental device and method Download PDFInfo
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- CN110116028A CN110116028A CN201910449523.7A CN201910449523A CN110116028A CN 110116028 A CN110116028 A CN 110116028A CN 201910449523 A CN201910449523 A CN 201910449523A CN 110116028 A CN110116028 A CN 110116028A
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- 238000000034 method Methods 0.000 title abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 99
- 238000002474 experimental method Methods 0.000 claims abstract description 52
- 230000003139 buffering effect Effects 0.000 claims abstract description 48
- 238000002360 preparation method Methods 0.000 claims abstract description 14
- 238000012544 monitoring process Methods 0.000 claims abstract description 12
- 239000012530 fluid Substances 0.000 claims description 13
- 230000002572 peristaltic effect Effects 0.000 claims description 13
- 239000000126 substance Substances 0.000 claims description 10
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 7
- 229910052760 oxygen Inorganic materials 0.000 claims description 7
- 239000001301 oxygen Substances 0.000 claims description 7
- 238000012360 testing method Methods 0.000 claims description 7
- 238000012546 transfer Methods 0.000 claims description 5
- 230000000052 comparative effect Effects 0.000 abstract description 7
- 230000007774 longterm Effects 0.000 abstract description 6
- 238000004904 shortening Methods 0.000 abstract 1
- 238000010586 diagram Methods 0.000 description 3
- 238000006073 displacement reaction Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 2
- 238000004377 microelectronic Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003739 neck Anatomy 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
Abstract
It includes micro-fluidic laboratory that the present invention, which provides a kind of microfluidic experimental device and method, described device, further includes by the sequentially connected stoste pushing equipment of microfluidic channel, liquid mixing bottle, the first microfluid pushing equipment, Multi-channel switching valve；The liquid outlet of the Multi-channel switching valve is connected with first-level buffer bottle and level 2 buffering bottle respectively, and the first-level buffer bottle and level 2 buffering bottle pass through the second microfluid pushing equipment respectively and be connected with the micro-fluidic laboratory.The present invention can satisfy the demand of multichannel comparative experiments, required time when shortening multiple groups experiment, and the long-term real-time monitoring to a variety of microenvironment parameters may be implemented, while reducing manual operation, improve the control precision of solution preparation and the stability of experiment.
Description
Technical field
The present invention relates to the emerging interleaving techniques of fluid physics, biology, chemistry, microelectronics and biomedical engineering etc. necks
Domain, specifically, being related to a kind of microfluidic experimental device and method.
Background technique
Micro-fluidic (microfluidics) is referred to using microchannel (having a size of tens of to hundreds of microns) processing or manipulation
Science and Technology involved in the system of minute fluid is one and is related to chemistry, fluid physics, microelectronics, new material, biology
With the emerging cross discipline of biomedical engineering.Publication No. CN108626102A Chinese patent literature discloses a kind of micro-fluidic
Device, including displacement actuator and micro-fluidic chip, institute's displacement actuator and the micro-fluidic chip are mutually indepedent；It is described micro-
Fluidic chip have microchannel, the microchannel have input terminal and output end, the input terminal of the microchannel with it is described
There is press section between output end；Institute's displacement actuator has the brake component for squeezing the press section microchannel.It is micro- with this
Flow control apparatus be representative it is existing based on micro-fluidic chip biological culture experiment have the following problems: (1) be unable to satisfy for
Multi-chip or same chip multi-chamber the comparative experiments especially demand of comparison simultaneously, and when carrying out multiple groups experiment the time required to compared with
It is long；(2) it cannot achieve and carry out long-term dynamic essence to experiment macro (nutritive salt, pH value etc.), physics (temperature, salinity) environment
Really control；(3) traditional micro-fluidic experiment or cell culture experiments are that manual operation is completed individually to match liquid and push with liquid, are not had
There is realization to automate, there are the artificial disturbing factor such as manually-operated control accuracy difference, stability is poor.
Summary of the invention
An object of the present invention is to solve existing in the prior art can not compare to test especially while comparing
And carry out required time longer problem when multiple groups experiment.
The second object of the present invention is to realize the long-term real-time monitoring to a variety of microenvironment parameters, while reducing artificial behaviour
Make, improves the control precision and stability of control system.
To achieve the above object, it present invention firstly provides a kind of microfluidic experimental device, including micro-fluidic laboratory, also wraps
It includes through the sequentially connected stoste pushing equipment of microfluidic channel, liquid mixing bottle, the first microfluid pushing equipment, multichannel switching
Valve；The liquid outlet of the Multi-channel switching valve is connected with first-level buffer bottle and level 2 buffering bottle respectively, the first-level buffer bottle and
Level 2 buffering bottle passes through the second microfluid pushing equipment respectively and is connected with the micro-fluidic laboratory.
It preferably, further include a controller, the stoste pushing equipment, the first microfluid pushing equipment, the second microfluid
Pushing equipment, Multi-channel switching valve are connected with the controller respectively.
Preferably, sensor is equipped in the liquid mixing bottle, the sensor is connected with the controller.
Preferably, the quantity of the stoste pushing equipment is N number of and is set side by side, N >=1 that each stoste pushing equipment is equal
It is connected respectively with the liquid mixing bottle.
Preferably, the quantity in the micro-fluidic laboratory is M and is set side by side, M >=1；The number of the first-level buffer bottle
Amount is similarly M, and the liquid outlet of each first-level buffer bottle passes through the channel a M conveying equipment for fluid substances and corresponding micro-fluidic experiment
Room is connected；The quantity of the level 2 buffering bottle is similarly M, and it is logical that the liquid outlet of each level 2 buffering bottle passes through another M
Road conveying equipment for fluid substances is connected with corresponding micro-fluidic laboratory.
Preferably, the micro-fluidic laboratory is micro-fluidic chip or biological culture dish or biologic test case.
Preferably, the stoste pushing equipment includes stoste bottle and the third pushing equipment being attached thereto.
Preferably, the sensor is one of pH sensor, salinity sensor, dissolved oxygen sensor or a variety of.
Preferably, the microfluid pushing equipment is one of pulsometer, peristaltic pump.
The present invention also provides a kind of micro-fluidic experimental methods based on above-mentioned experimental provision, include the following steps:
Step 1: respective standard liquid needed for being respectively charged into experiment into N number of stoste pushing equipment；
Step 2: controller controls the stoste pushing equipment to push to respective standard liquid in liquid mixing bottle, matches simultaneously
The respective concentration signal of solution and this concentration signal is sent to controller in sensor real-time monitoring liquid mixing bottle in liquid bottle；
Step 3: whether the concentration signal and actually required concentration signal that controller judgement receives are consistent, if consistent
Stop stoste pushing equipment to liquid mixing bottle transfer criteria liquid, completes first time solution and prepare；Stoste is continued through if inconsistent
Pushing equipment to liquid mixing bottle conveying respective standard liquid until all concentration signals it is consistent with actually required concentration signal, completion first
Secondary solution is prepared；
Step 4:, will through Multi-channel switching valve by the first microfluid conveying equipment after the completion of the preparation of first time solution
Solution in liquid mixing bottle pushes in first first-level buffer bottle；
Step 5: repeating step 2 and step 3, is sequentially completed that second of solution is prepared, third time solution is prepared ... the
M times solution is prepared, and corresponding solution is accordingly pushed to second first-level buffer bottle, third first-level buffer bottle ... m-th
First-level buffer bottle；
Step 6:, will be molten in this M first-level buffer bottle after completing corresponding solution push in M first-level buffer bottle
Liquid pushes in corresponding micro-fluidic laboratory simultaneously and carries out first group of experiment；
Step 7: while carrying out step 6, step 2 and step 3 is repeated, is respectively completed needed for next group of experiment
The M time solution preparation that first time solution is prepared, second solution is prepared, third time solution is prepared ..., and by corresponding solution phase
First level 2 buffering bottle, second level 2 buffering bottle, third level 2 buffering bottle ... m-th level 2 buffering should be pushed to
Bottle is waited after completing corresponding solution push in M level 2 buffering bottle and being operated in next step；
Step 8: after the completion of first group of experiment, the solution in M level 2 buffering bottle is pushed into corresponding miniflow simultaneously
It controls and carries out second group of experiment in laboratory.
The positive effect of the present invention:
1. for the consistency for guaranteeing the time, add between liquid mixing bottle and micro-fluidic laboratory (or same with laboratory quantity
The chamber quantity in one laboratory) equivalent amount first-level buffer bottle, simultaneously to micro-fluidic laboratory after complete soln configures
Interior push.
2. when solution is pushed into chip in first-level buffer bottle, can continue to configure molten to save the time cost for matching liquid
Liquid is stored in level 2 buffering bottle, and time-sharing multiplex improves working efficiency.
3. the pushing equipment that solution pushes liquid into I and II surge flask in liquid mixing bottle only needs a channel, and
Channel automatic transfer valve is added, after the pushing equipment to complete to match liquid to multiple surge flasks.
4. can satisfy the mixing of multiple standards liquid with liquid, the multiple microenvironment parameters of real-time control.
5. it is carried out using corresponding microfluid pushing equipment with liquid during prolonged observation or culture experiment, it can
To save the artificial process for matching liquid, and will not because of syringe specification problem and influence the precision of control.
6. solving the fine difference when doing comparative experiments due to solution in proportion by setting liquid mixing bottle and preparing
Complicated problem is that is, accurate using same set of equipment and be simply done the mixed preparing of different solutions.
7. associated micro fluidic pushing equipment is selectively wide, it may be employed without limitation of the equipment such as peristaltic pump, pulsometer.
8. forming the closed loop regulation that solution is prepared by controller and sensor feedback, the real-time control of solution preparation is realized
System, and the precision of solution preparation has been effectively ensured, i.e., traditional biological culture environment conditioning based on culture dish or chamber
Control is mainly human eye observation, manually allotment etc., and required microfluid is small in size in micro-fluidic laboratory, thus dense in liquid
Requirement on degree is higher, and being equipped with high-precision sensor using the present invention at this time may be implemented real-time high-precision closed loop, such as
PH: ± 0.01；Salinity: ± 0.1ppt；Dissolved oxygen: ± 0.1mg/L.
9. surge flask and liquid mixing bottle are able to maintain good leakproofness, to reduce solubility variation caused by solution evaporates, together
When can meet using pulsometer etc. as air pressure requirement when pushing equipment.
In short, the present invention may be implemented to can satisfy multi-pass trace comparison to the long-term real-time monitoring of a variety of microenvironment parameters
The demand that experiment especially compares simultaneously, while reducing manual operation, improve the control precision and experiment of solution preparation
Stability.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 is the attachment structure schematic diagram of controller of the present invention Yu each component；
Fig. 2 is the structural schematic diagram of microfluidic experimental device of the present invention；
Fig. 3 is the structural schematic diagram of microfluidic experimental device described in embodiment 1.
Specific embodiment
Below in conjunction with attached drawing of the invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
In addition, it should be noted that, in the description of the present invention, term "left", "right", "upper", "lower", " front and back " etc. refer to
The orientation or positional relationship shown is to be based on the orientation or positional relationship shown in the drawings, and is merely for convenience of description this product and simplification
Description does not have Stated otherwise such as, and it is not the device or element of indication or suggestion meaning that there is no particular meanings for above-mentioned word
It must have a particular orientation, be constructed and operated in a specific orientation, therefore should not be understood as the limitation to this patent.
According to an aspect of the present invention, referring to figs. 1 to Fig. 2, the present invention provides a kind of microfluidic experimental device, including micro-
Flow control laboratory 12 further includes being pushed away by the sequentially connected stoste pushing equipment of microfluidic channel, liquid mixing bottle 9, the first microfluid
Send equipment 4, Multi-channel switching valve 7；The liquid outlet of the Multi-channel switching valve 7 respectively with first-level buffer bottle 10 and level 2 buffering bottle
11 are connected, the first-level buffer bottle 10 and level 2 buffering bottle 11 respectively by the second microfluid pushing equipment 5 with it is described micro-fluidic
Laboratory 12 is connected.By being arranged between liquid mixing bottle 9 and micro-fluidic laboratory 12 and micro-fluidic laboratory quantity (or same reality
Test the chamber quantity of room) the first-level buffer bottle of equivalent amount, it can be after complete soln configures while into micro-fluidic laboratory
Push, to realize comparative experiments and be to compare simultaneously, and when solution is pushed into chip in first-level buffer bottle, can continue to match
Solution deposit level 2 buffering bottle is set, time-sharing multiplex saves the time with liquid, improves work efficiency.
Further, to reduce manual operation, the stability of control precision and experiment that solution is prepared, the miniflow are improved
Control experimental provision further includes a controller 2, the stoste pushing equipment, the first microfluid pushing equipment 4, the push of the second microfluid
Equipment 5, Multi-channel switching valve 7 are connected with the controller 2 respectively.Preferably, for convenient for control operation, the controller 2 is also
It is connected with a host computer 1.
Preferably, to realize the long-term real-time monitoring to a variety of microenvironment parameters, sensor is additionally provided in the liquid mixing bottle 9
6, the sensor 6 is connected with the controller 2.The real-time height of microenvironment may be implemented using the configuration of this high-precision sensor
The regulation of precision closed loop, such as pH: ± 0.01；Salinity: ± 0.1ppt；Dissolved oxygen: ± 0.1mg/L.
Preferably, according to actually required, the micro-fluidic laboratory 12 can for micro-fluidic chip or biological culture dish or
The test site such as biologic test case.
Preferably, the stoste pushing equipment includes stoste bottle 8 and the third pushing equipment 3 being attached thereto, the third
Pushing equipment 3 is connected with the controller 2.Certainly, the stoste pushing equipment can also be that collection stoste bottle and third push are set
Standby function is in the equipment such as the syringe pump of one or plunger pump.
Preferably, the sensor 6 includes but is not limited to pH sensor, salinity sensor, one in dissolved oxygen sensor
Kind is a variety of, and the measuring probe of sensor is placed in liquid mixing bottle, and the survey to configuration microenvironment parameter (PH, salinity etc.) may be implemented
Amount.
Preferably, microfluid pushing equipment (the first microfluid pushing equipment, the second microfluid pushing equipment, third
Microfluid pushing equipment) it is one of pulsometer, peristaltic pump, plunger pump, syringe pump or a variety of is used cooperatively.
Specifically, on the one hand the stoste pushing equipment is used to hold different titers, that is, need needed for holding experiment
The original sample titer to be configured according to different proportion, such as PH titer, salinity standard liquid.The quantity of the stoste pushing equipment
Be it is N number of and be set side by side, N >=1, each stoste pushing equipment is connected with liquid mixing bottle 9 respectively.
The liquid mixing bottle 9 is the place that each titer is mixed according to different proportion.
Further, the micro-fluidic laboratory 12 is experimental place, can be single micro-fluidic if wanting to complete comparative experiments
The multiple chambers of indoor design are tested, or complete comparative experiments using multiple micro-fluidic laboratories, at this to micro-fluidic laboratory
Pattern or material are not required.Specifically, to compare experiment, the quantity in the micro-fluidic laboratory 12 can it is a for M (or
The chamber quantity in the single micro-fluidic laboratory of person is M) and be set side by side, M >=1；The quantity of the first-level buffer bottle 10 is similarly M
It is a, the liquid outlet of each first-level buffer bottle 10 pass through the channel a M conveying equipment for fluid substances (third microfluid pushing equipment 5) with
Corresponding micro-fluidic laboratory 12 is connected；The quantity of the level 2 buffering bottle 11 is similarly M, and each level 2 buffering bottle 11 goes out liquid
Mouth passes through another channel M conveying equipment for fluid substances (another third microfluid pushing equipment 5) and corresponding micro-fluidic experiment
Room 12 is connected.
The surge flask (first-level buffer bottle 10 and level 2 buffering bottle 11) is the places to be conveyed such as the solution that configuration is completed,
It is pushed in corresponding micro-fluidic laboratory 12 simultaneously after the completion of being prepared convenient for all solution.
The microfluidic channel is for connecting composition component (stoste bottle, liquid mixing bottle, surge flask, a micro-fluidic laboratory
Deng), material is unlimited, can be silicone tube etc..
Specifically, the another aspect effect of the stoste pushing equipment is for completing titer pushing away to liquid mixing bottle
It send；The first microfluid pushing equipment is for completing push of the liquid mixing bottle to liquid between surge flask；Second microfluid pushes away
Send equipment for completing push of the surge flask to corresponding micro-fluidic experiment indoor liquid.Each microfluid conveying equipment can be plunger
One of pump, pulsometer, peristaltic pump, syringe pump.
The Multi-channel switching valve is arranged between liquid mixing bottle and surge flask and after the first microfluid pushing equipment, uses
Different pipelines is switched to by the switching valve in the solution for transferring out the first microfluid conveying equipment, the same time only has one
A valve pipe conducting.
According to another aspect of the present invention, the micro-fluidic experimental method based on above-mentioned apparatus that the present invention also provides a kind of,
Include the following steps:
Step 1: respective standard liquid (such as PH standard needed for being respectively charged into experiment into N number of stoste pushing equipment
Liquid, salinity standard liquid, basal liquid etc.)；
Step 2: controller controls the stoste pushing equipment will with (runing time and the speed of service that control this equipment)
Respective standard liquid pushes in liquid mixing bottle, while the respective concentration of solution is believed in the sensor real-time monitoring liquid mixing bottle in liquid mixing bottle
Number and this concentration signal is sent to controller；
Step 3: whether the concentration signal and actually required concentration signal that controller judgement receives are consistent, if consistent
Stop stoste pushing equipment to liquid mixing bottle transfer criteria liquid, completes first time solution and prepare；Stoste is continued through if inconsistent
Pushing equipment to liquid mixing bottle conveying respective standard liquid until all concentration signals it is consistent with actually required concentration signal, completion first
Secondary solution is prepared；
Step 4:, will through Multi-channel switching valve by the first microfluid conveying equipment after the completion of the preparation of first time solution
Solution in liquid mixing bottle pushes in first first-level buffer bottle；
Step 5: repeating step 2 and step 3, is sequentially completed that second of solution is prepared, third time solution is prepared ... the
M times solution is prepared, and corresponding solution is accordingly pushed to second first-level buffer bottle, third first-level buffer bottle ... m-th
First-level buffer bottle；
Step 6:, will be molten in this M first-level buffer bottle after completing corresponding solution push in M first-level buffer bottle
Liquid is pushed in corresponding micro-fluidic laboratory simultaneously by second pushing equipment in a channel M and carries out first group of experiment；
Step 7: while carrying out step 6, step 2 and step 3 is repeated, is respectively completed needed for next group of experiment
The M time solution preparation that first time solution is prepared, second solution is prepared, third time solution is prepared ..., and by corresponding solution phase
First level 2 buffering bottle, second level 2 buffering bottle, third level 2 buffering bottle ... m-th level 2 buffering should be pushed to
Bottle is waited after completing corresponding solution push in M level 2 buffering bottle and being operated in next step；
Step 8: after the completion of first group of experiment, the solution in M level 2 buffering bottle is passed through into another channel M
Second pushing equipment pushes in corresponding micro-fluidic laboratory simultaneously and carries out second group of experiment.
Hereafter detailed illustration will be carried out to the experimental provision by specific embodiment 1 and with reference to attached drawing 3.
Embodiment 1
Referring to Fig. 3, it further includes passing through that the present embodiment 1, which provides a kind of microfluidic experimental device, including micro-fluidic laboratory 12,
The sequentially connected stoste pushing equipment of microfluidic channel, liquid mixing bottle 9, the first microfluid pushing equipment 4, Multi-channel switching valve 7；Institute
The liquid outlet for stating Multi-channel switching valve 7 is connected with first-level buffer bottle 10 and level 2 buffering bottle 11 respectively, the first-level buffer bottle 10
It is connected respectively by the second microfluid pushing equipment 5 with the micro-fluidic laboratory 12 with level 2 buffering bottle 11.
Further, the microfluidic experimental device further includes a controller 2, the stoste pushing equipment, the first miniflow
Body pushing equipment 4, the second microfluid pushing equipment 5, Multi-channel switching valve 7 are connected with the controller 2 respectively, the controller
2 are also connected with a host computer 1.
Further, sensor 6 is additionally provided in the liquid mixing bottle 9, the sensor 6 is connected with the controller 2.
Further, the micro-fluidic laboratory 12 is micro-fluidic chip.
Further, the stoste pushing equipment includes stoste bottle 8 and the third pushing equipment 3 being attached thereto, and described
Three pushing equipments 3 are connected with the controller 2.
Further, the sensor 6 is pH sensor, salinity sensor, the whole in dissolved oxygen sensor.
Further, the microfluid pushing equipment is peristaltic pump.
Specifically, the quantity of the stoste bottle is four (respectively stoste bottle A, stoste bottle B, stoste bottle C, stoste bottle D)
And be set side by side, each stoste bottle passes through single channel peristaltic pump (the first microfluid conveying equipment) and liquid mixing bottle phase
Even.
To compare experiment, the quantity of the micro-fluidic chip is four (respectively micro-fluidic chip A, micro-fluidic core
Piece B, micro-fluidic chip C, micro-fluidic chip D) and be set side by side；The quantity of the first-level buffer bottle is similarly four (respectively
First-level buffer bottle A, first-level buffer bottle B, first-level buffer bottle C, first-level buffer bottle D), the liquid outlet of each first-level buffer bottle passes through
One four-way peristaltic pump (the second microfluid pushing equipment) is connected with corresponding micro-fluidic chip；The quantity of the level 2 buffering bottle
It is similarly four (respectively level 2 buffering bottle A, level 2 buffering bottle B, level 2 buffering bottle C, level 2 buffering bottle D), each second level is slow
The liquid outlet for rushing bottle passes through another four-way conveying equipment for fluid substances (another the second microfluid pushing equipment) and corresponding
Micro-fluidic chip is connected.
Correspondingly, the first microfluid conveying equipment is single channel peristaltic pump, the Multi-channel switching valve is eight channels
Switching valve (number of channels of switching valve be micro-fluidic chip quantity twice).
The present embodiment 1 also provides a kind of micro-fluidic experimental method based on above-mentioned experimental provision, includes the following steps:
Step 1: respective standard liquid (such as PH titer, salinity needed for being respectively charged into experiment into four stoste bottles
Titer, basal liquid etc.)；
Step 2: controller controls the third conveying equipment for fluid substances (corresponding single channel peristaltic pump) with by corresponding stoste bottle
In titer push in liquid mixing bottle, while in the sensor real-time monitoring liquid mixing bottle in liquid mixing bottle solution respective concentration letter
Number and this concentration signal is sent to controller；
Step 3: whether the concentration signal and actually required concentration signal that controller judgement receives are consistent, if consistent
Stop third conveying equipment for fluid substances to liquid mixing bottle transfer criteria liquid, completes first time solution and prepare；It is continued through if inconsistent
Third conveying equipment for fluid substances to liquid mixing bottle conveying respective standard liquid until all concentration signals it is consistent with actually required concentration signal,
First time solution is completed to prepare；
Step 4: it after the completion of the preparation of first time solution, is passed through by the first microfluid conveying equipment (single channel peristaltic pump)
Eight channel switching valves push to the solution in liquid mixing bottle in first-level buffer bottle A；
Step 5: repeating step 2 and step 3, is sequentially completed second of solution preparation, the preparation of third time solution, the 4th
Secondary solution is prepared, and corresponding solution is accordingly pushed to first-level buffer bottle B, first-level buffer bottle C, first-level buffer bottle D；
Step 6:, will be in this four first-level buffer bottles after completing corresponding solution push in four first-level buffer bottles
Solution is pushed to simultaneously by four-way peristaltic pump and carries out first group of experiment in corresponding micro-fluidic chip；
Step 7: while carrying out step 6, step 2 and step 3 is repeated, is respectively completed needed for next group of experiment
First time solution is prepared, second of solution is prepared, third time solution is prepared, the 4th solution is prepared, and corresponding solution is pushed
To level 2 buffering bottle A, level 2 buffering bottle B, level 2 buffering bottle C, level 2 buffering bottle D, to complete phase in four level 2 buffering bottles
It waits after answering solution to push and operating in next step；
Step 8: after the completion of first group of experiment, the solution in four level 2 buffering bottles is passed through into another four-way
Peristaltic pump pushes to simultaneously carries out second group of experiment in corresponding micro-fluidic chip.
The present embodiment may be implemented that (microenvironment real-time monitoring refers to micro- to the long-term real-time monitorings of a variety of microenvironment parameters
The intrinsic parameters such as the salinity in microenvironment, dissolved oxygen, PH in fluidic chip carry out real-time monitoring, create suitable biological growth or
Accurate microenvironment needed for experiment can create a biological culture research platform, for seeing by microenvironment real-time monitoring
Survey and cultivate biological growth conditions and growth mechanism), and can satisfy multichannel comparative experiments especially while comparing
Demand effectively shortens the time required when multiple groups experiment, while reducing manual operation, improves the control essence of solution preparation
The stability of degree and experiment.
It is above-described to be merely a preferred embodiment of the present invention, it should be understood that the explanation of above embodiments is only used
In facilitating the understanding of the method and its core concept of the invention, it is not intended to limit the scope of protection of the present invention, it is all of the invention
Any modification for being made within thought and principle, equivalent replacement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of microfluidic experimental device, including micro-fluidic laboratory, it is characterised in that: further include by microfluidic channel successively
Stoste pushing equipment, liquid mixing bottle, the first microfluid pushing equipment, Multi-channel switching valve of connection；The Multi-channel switching valve
Liquid outlet is connected with first-level buffer bottle and level 2 buffering bottle respectively, and the first-level buffer bottle and level 2 buffering bottle pass through second respectively
Microfluid pushing equipment is connected with the micro-fluidic laboratory.
2. a kind of microfluidic experimental device according to claim 1, it is characterised in that: further include a controller, the original
Liquid pushing equipment, the first microfluid pushing equipment, the second microfluid pushing equipment, Multi-channel switching valve respectively with the controller
It is connected.
3. a kind of microfluidic experimental device according to claim 2, it is characterised in that: be equipped with sensing in the liquid mixing bottle
Device, the sensor are connected with the controller.
4. a kind of microfluidic experimental device according to claim 1 or 2 or 3, it is characterised in that: the stoste pushing equipment
Quantity be it is N number of and be set side by side, N >=1, each stoste pushing equipment is connected with the liquid mixing bottle respectively.
5. a kind of microfluidic experimental device according to claim 1 or 2 or 3, it is characterised in that: the micro-fluidic laboratory
Quantity be M and be set side by side, M >=1；The quantity of the first-level buffer bottle is similarly M, and each first-level buffer bottle goes out liquid
Mouth is connected by the channel a M conveying equipment for fluid substances with corresponding micro-fluidic laboratory；The quantity of the level 2 buffering bottle is same
It is M, the liquid outlet of each level 2 buffering bottle passes through another channel M conveying equipment for fluid substances and corresponding micro-fluidic laboratory
It is connected.
6. a kind of microfluidic experimental device according to claim 1 or 2 or 3, it is characterised in that: the micro-fluidic laboratory
For micro-fluidic chip or biological culture dish or biologic test case.
7. a kind of microfluidic experimental device according to claim 1 or 2 or 3, it is characterised in that: the stoste pushing equipment
Including stoste bottle and the third pushing equipment being attached thereto.
8. a kind of microfluidic experimental device according to claim 3, it is characterised in that: the sensor be pH sensor,
One of salinity sensor, dissolved oxygen sensor are a variety of.
9. a kind of microfluidic experimental device according to claim 1 or 2 or 3, it is characterised in that: the microfluid push is set
Standby is one of pulsometer, peristaltic pump.
10. a kind of micro-fluidic experimental method based on microfluidic experimental device described in claim 1-9, which is characterized in that including
Following steps:
Step 1: respective standard liquid needed for being respectively charged into experiment into N number of stoste pushing equipment；
Step 2: controller controls the stoste pushing equipment to push to respective standard liquid in liquid mixing bottle, while liquid mixing bottle
In sensor real-time monitoring liquid mixing bottle in solution respective concentration signal and this concentration signal is sent to controller；
Step 3: whether the concentration signal and actually required concentration signal that controller judgement receives are consistent, stop if consistent
Stoste pushing equipment is completed first time solution and is prepared to liquid mixing bottle transfer criteria liquid；Stoste push is continued through if inconsistent
For equipment to liquid mixing bottle conveying respective standard liquid until all concentration signals are consistent with actually required concentration signal, completion is molten for the first time
Liquid is prepared；
Step 4: after the completion of the preparation of first time solution, liquid will be matched through Multi-channel switching valve by the first microfluid conveying equipment
Solution in bottle pushes in first first-level buffer bottle；
Step 5: repeating step 2 and step 3, is sequentially completed second solution preparation, third time solution and prepares ... the M times
Solution is prepared, and corresponding solution is accordingly pushed to second first-level buffer bottle, third first-level buffer bottle ... m-th level-one
Surge flask；
Step 6: after completing corresponding solution push in M first-level buffer bottle, the solution in this M first-level buffer bottle is same
When push to first group of experiment of progress in corresponding micro-fluidic laboratory；
Step 7: while carrying out step 6, step 2 and step 3 is repeated, is respectively completed first needed for next group of experiment
Secondary solution is prepared, second of solution is prepared, third time solution is prepared ..., and the M times solution is prepared, and corresponding solution is accordingly pushed away
It send to first level 2 buffering bottle, second level 2 buffering bottle, third level 2 buffering bottle ... m-th level 2 buffering bottle, to M
Operation in next step is waited after completing corresponding solution push in a level 2 buffering bottle；
Step 8: after the completion of first group of experiment, the solution in M level 2 buffering bottle is pushed to corresponding micro-fluidic reality simultaneously
Test second group of experiment of indoor carry out.
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