CN109647547A - A kind of preparation method of the controllable aqueous two-phase drop based on microflow control technique - Google Patents
A kind of preparation method of the controllable aqueous two-phase drop based on microflow control technique Download PDFInfo
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- 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
- B01L3/502769—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 characterised by multiphase flow arrangements
- B01L3/502784—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 characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
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Abstract
The preparation method of the invention discloses a kind of controllable aqueous two-phase drop based on microflow control technique.This method includes the integrated of micro-fluidic pump valve system, double-aqueous phase system selection, regulation of drop etc..The present invention using can the separation of spontaneous phase double-aqueous phase system, be integrated at one in " cross " the flow liquid drop chip of pump valve system, stablized by adjusting two-phase flow velocity, pump valve switch periods etc., controllably form drop.The technology is expected to play a role in the biological applications such as Separation of Proteins, Cellular compartment culture, DNA extraction.
Description
Technical field
The invention belongs to the fields such as microflow control technique, materials chemistry, and especially it is related to a kind of controllable based on microflow control technique
The preparation method of aqueous two-phase drop.
Background technique
Microfluidic droplet technology is a micro-fluidic important branch, it can directly using property incompatible between fluid come
Monodisperse drop is obtained, and the technology of manipulations such as sorted, divided, captured, merged to it.Microfluidic droplet has biggish
Specific surface area can efficiently complete mass exchange, chemical reaction etc. in small volume.Therefore, the technology is at small point
The fields such as sub- detection, single cell analysis, drug delivery, microparticle synthesis, organizational project are widely used.
However, traditional Microfluidic droplet technology is all based on the double newborn phase systems of grease to generate uniform controllable drop,
The use to organic phase and all kinds of ions or nonionic surfactant can be wherein related to, and these substances are difficult in system
It completely removes, this allows for application of the technology in field of biomedicine and is limited.In recent years, with traditional grease two-phase
The different double-aqueous phase system of system is introduced in Microfluidic droplet field, and has obtained preliminary development.As its name suggests, aqueous two-phase
Organic phase is not present in system, the principle mutually separated is, when two kinds of solute polymers of different nature in aqueous solution dense
More than after certain threshold value, the physicochemical properties of two-phase aqueous solution can also change degree therewith, to generate spontaneous phase point
From.Double-aqueous phase system is the extraction for biomass originally, therefore has good biocompatibility.The composition of aqueous two-phase can be with
It is the different Polymer Solution of two kinds of properties (such as PEG and glucan) or a kind of Polymer Solution and a kind of salting liquid (such as PEG
And phosphate), the system that two of them Polymer Solution is constituted has better biocompatibility.More common and phase point at present
It is made of from the preferable system of effect PEG and glucan.But double-aqueous phase system obtains drawback there is also obvious, due to two water
Alternate surface can will be much smaller than grease system, and the generation of aqueous two-phase drop, especially controllable generate just become this system
The widely applied obstruction in Microfluidic droplet technology.For the present invention using the micro-fluidic chip of pump valve system is integrated with, success is real
The preparation of uniform controllable aqueous two-phase drop is showed.
Summary of the invention
The purpose of the present invention is to provide one kind to be based on micro-fluidic chip pump valve integrated system, for controllably generating equal a pair of
The new method of aqueous phase droplets.
A kind of controllable aqueous two-phase drop preparation method based on microflow control technique of the present invention, including the following steps:
(1) pump valve system is integrated: using the method for conventional soft lithographic, PDMS chip is prepared, based on one traditional " ten
Word " stream Microfluidic droplet chip, the dispersed phase channel two sides integrated pneumatic power pump valve in " cross " intersection upstream,；The chip
Mainly by continuous phase entrance, gas access, dispersed phase entrance, drop outlets, dispersed phase channel, gas passage, continuous phase channel,
Main channel and pump valve composition；
Continuous phase entrance, dispersed phase entrance pass through continuous phase channel, dispersed phase channel respectively and connect with main channel, dispersed phase
Channel and continuous phase channel converge to formation " cross " intersection at main channel；Gas access reaches air driven pump through gas passage
Elastic deformation occurs for valve, gas-powered pump valve side wall therein；
The position of pump valve passes through pump valve inflation and two kinds of shapes of tranquillization in the dispersed phase channel two sides of " cross " intersection upstream
State periodically squeezes dispersed phase channel and forms aqueous two-phase stably and controllablely so that dispersed phase be made to enter in continuous phase by phased manner
Drop；
(2) double-aqueous phase system selects: the double-aqueous phase system used is the system either one of two Polymer Solutions composition
The system of kind Polymer Solution and a kind of salting liquid composition,
The system of described two Polymer Solution compositions is polyethylene glycol (PEG)-glucan, polypropylene glycol (PPG)-Portugal is poly-
Sugar, polyvinyl alcohol (PVA)-methylcellulose etc. can the separation of spontaneous phase combination；
The system of a kind of Polymer Solution and a kind of salting liquid composition is PEG- sodium sulphate, PPG- potassium phosphate, poly- second
Vinyl pyrrolidone (PVP)-potassium phosphate etc. can the separation of spontaneous phase combination；
Selecting double-aqueous phase system is the combination of PEG- glucan, the PEG molecular weight ranges 1000-20000Da, concentration
Range: 2.5-50%；Dextran molecule amount range: 70k-500kDa, concentration range: 2.5-20%.
(3) regulation of drop: using glucan as dispersed phase, PEG is as continuous phase；Or PEG is as dispersed phase, glucan
As continuous phase；Period by changing dispersed phase flow velocity, continuous phase flow velocity and pump valve switch forms drop to change in channel
Size and drop spacing.Dispersed phase flow rates: 0.01-1 μ l/min, continuous phase flow rates: 0.5-5 μ l/min,
Pump valve switch time 0.05-0.5s.
100-300 μm of width of the chip main channel, long 1-2cm.Pump valve and dispersed phase interchannel are away from 40-60 μm, pump valve
Between 40-60 μm of dispersed phase channel width, chip each section channel height is 100-300 μm；
It is very uniform to be formed by aqueous two-phase drop size, 30-300 μm of adjustable diameter range.It is expected to be applied to albumen
In the biological applications such as matter separation, Cellular compartment culture, DNA extraction.
The present invention using can the separation of spontaneous phase double-aqueous phase system, " cross " flow liquid of pump valve system is integrated at one
It drips in chip, is stablized by adjusting two-phase flow velocity, pump valve switch periods etc., controllably forms drop.The technology is expected in albumen
It plays a role in the biological applications such as matter separation, Cellular compartment culture, DNA extraction.
Detailed description of the invention
Fig. 1 is single channel aqueous two-phase drop chip schematic diagram, in which: a overall pattern；B pump valve structure partial figure；C pump valve knot
Structure switch state schematic diagram.
1 represents continuous phase entrance；2 represent gas access；3 represent dispersed phase entrance；4 represent drop outlets；5 represent dispersion
Phase channel；6 represent gas passage；7 represent continuous phase channel；8 represent main channel；9 represent pneumatic pump valve.
Fig. 2 is the aqueous two-phase drop pictorial diagram and diameter distribution statistics figure of embodiment 1, wherein a drop pictorial diagram (scale:
200μm)；B diameter statistical chart.
Fig. 3 is the aqueous two-phase drop pictorial diagram and diameter distribution statistics figure of embodiment 2, wherein a drop pictorial diagram (scale:
200μm)；B diameter statistical chart.
Specific embodiment
First according to actual needs, the Microfluidic droplet chip that design and processing dimension are suitable for；Then double water that selection needs
Phase composition system, including type, concentration, molecular weight etc.；Finally utilize dispersed phase flow velocity, continuous phase flow velocity and pump valve switching frequency
Drop parameter can be regulated and controled.Present invention will be further explained below with reference to the attached drawings and examples.
Embodiment 1
A kind of controllable aqueous two-phase drop preparation method based on microflow control technique, it is characterised in that: include the following steps:
(1) pump valve system is integrated: using the method for conventional soft lithographic, PDMS chip is prepared, based on one traditional " ten
Word " flows Microfluidic droplet chip, the dispersed phase channel two sides integrated pneumatic power pump valve in " cross " intersection upstream；The chip is such as
Shown in Fig. 1, mainly by continuous phase entrance 1, gas access 2, dispersed phase entrance 3, drop outlets 4, dispersed phase channel 5, gas is logical
Road 6, continuous phase channel 7, main channel 8 and pump valve 9 form；
Continuous phase entrance 1, dispersed phase entrance 3 are connect by continuous phase channel 7, dispersed phase channel 5 with main channel 8 respectively,
Dispersed phase channel and continuous phase channel converge to formation " cross " intersection at main channel；Gas access 2 is reached through gas passage 6
Elastic deformation occurs for pneumatic pump valve 9, gas-powered pump valve side wall therein；
The position of pump valve 9 passes through pump valve inflation and two kinds of tranquillization in 5 two sides of dispersed phase channel of " cross " intersection upstream
Period of state squeezes dispersed phase channel, thus enter dispersed phase in continuous phase by phased manner, stably and controllable landform water in pairs
Phase drop；
200 μm of the chip main channel width, long 1.5cm.Point of pump valve and dispersed phase interchannel away from 50 μm, between pump valve
40 μm of dephasing channel width, chip each section channel height is 100-300 μm；
(2) double-aqueous phase system selects: using the combination of PEG- glucan.PEG molecular weight is 20kDa, concentration 17%；Portugal
Glycan molecule is 500kDa, concentration 15%.
(3) regulation of drop: using glucan as dispersed phase, PEG is as continuous phase；By changing dispersed phase flow velocity, connecting
The period of continuous phase flow velocity and pump valve switch changes the size and drop spacing that drop is formed in channel.Dispersed phase flow velocity is
0.2 μ l/min, continuous phase flow velocity are 2 μ l/min, pump valve switch time 0.2s.To the experimental data of collection, it is straight drop has been counted
Diameter, result are 88.0 ± 3.5 μm, as shown in Figure 2.
Embodiment 2
(1) pump valve system is integrated: using the method for conventional soft lithographic, PDMS chip is prepared, based on one traditional " ten
Word " flows Microfluidic droplet chip, the dispersed phase channel two sides integrated pneumatic power pump valve in " cross " intersection upstream；The chip is such as
Shown in Fig. 1, mainly by continuous phase entrance 1, gas access 2, dispersed phase entrance 3, drop outlets 4, dispersed phase channel 5, gas is logical
Road 6, continuous phase channel 7, main channel 8 and pump valve 9 form；
Continuous phase entrance 1, dispersed phase entrance 3 are connect by continuous phase channel 7, dispersed phase channel 5 with main channel 8 respectively,
Dispersed phase channel and continuous phase channel converge to formation " cross " intersection at main channel；Gas access 2 is reached through gas passage 6
Elastic deformation occurs for pneumatic pump valve 9, gas-powered pump valve side wall therein；
The position of pump valve 9 passes through pump valve inflation and tranquillization two in dispersed phase channel (5) two sides of " cross " intersection upstream
Kind period of state squeezes dispersed phase channel, so that dispersed phase be made to enter in continuous phase by phased manner, stably and controllable landform is in pairs
Aqueous phase droplets；
200 μm of the chip main channel width, long 1.5cm.Point of pump valve and dispersed phase interchannel away from 50 μm, between pump valve
40 μm of dephasing channel width, chip each section channel height is 200 μm；
(2) double-aqueous phase system selects: using the combination of PEG- glucan.PEG molecular weight is 8kDa, concentration 10%；Portugal
Glycan molecule is 70kDa, concentration 10%.
(3) regulation of drop: using glucan as dispersed phase, PEG is as continuous phase；By changing dispersed phase flow velocity, connecting
The period of continuous phase flow velocity and pump valve switch changes the size and drop spacing that drop is formed in channel.Dispersed phase flow velocity is
0.1 μ l/min, continuous phase flow velocity are 2 μ l/min, pump valve switch time 0.4s.To the experimental data of collection, it is straight drop has been counted
Diameter, result are 88.0 ± 3.5 μm, as shown in Figure 3.
Claims (3)
1. a kind of preparation method of the controllable aqueous two-phase drop based on microflow control technique, it is characterised in that: include the following steps:
(1) pump valve system is integrated: using the method for conventional soft lithographic, PDMS chip is prepared, based on one traditional " cross "
Stream Microfluidic droplet chip, the dispersed phase channel two sides integrated pneumatic power pump valve in " cross " intersection upstream,；The chip is main
By continuous phase entrance (1), gas access (2), dispersed phase entrance (3), drop outlets (4), dispersed phase channel (5), gas passage
(6), continuous phase channel (7), main channel (8) and pump valve (9) composition；
Continuous phase entrance (1), dispersed phase entrance (3) pass through continuous phase channel (7), dispersed phase channel (5) and main channel (8) respectively
Connection, dispersed phase channel and continuous phase channel converge to formation " cross " intersection at main channel；Gas access (2) is logical through gas
Road (6) reaches pneumatic pump valve (9), and elastic deformation occurs for gas-powered pump valve side wall therein；
The position of pump valve (9) passes through pump valve inflation and two kinds of tranquillization in dispersed phase channel (5) two sides of " cross " intersection upstream
Period of state squeezes dispersed phase channel, thus enter dispersed phase in continuous phase by phased manner, stably and controllable landform water in pairs
Phase drop,
(2) double-aqueous phase system selects: selecting double-aqueous phase system for the combination of PEG- glucan；The PEG molecular weight ranges:
1000-20000Da, concentration range: 2.5-50%；Dextran molecule amount range: 70k-500kDa, concentration range: 2.5-20%；
(3) regulation of drop: using glucan as dispersed phase, PEG is as continuous phase or PEG as dispersed phase, glucan conduct
Continuous phase；Change the ruler that drop is formed in channel by changing the period of dispersed phase flow velocity, continuous phase flow velocity and pump valve switch
Very little size and drop spacing；Dispersed phase flow rates: 0.01-1 μ l/min, continuous phase flow rates: 0.5-5 μ l/min, pump valve
Switch time 0.05-0.5s.
2. a kind of preparation method of controllable aqueous two-phase drop based on microflow control technique according to claim 1, feature
It is: 100-300 μm of width of the chip main channel, long 1-2cm；Pump valve and dispersed phase interchannel are away from 40-60 μm, between pump valve
40-60 μm of dispersed phase channel width, chip each section channel height is 100-300 μm.
3. a kind of preparation method of controllable aqueous two-phase drop based on microflow control technique according to claim 1, feature
Be: the aqueous two-phase drop size of formation is very uniform, 30-300 μm of adjustable diameter range；It is expected to applied to protein point
From in the biological applications such as, Cellular compartment culture, DNA extraction.
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| CN110841734A (en) * | 2019-12-06 | 2020-02-28 | 中国科学院长春光学精密机械与物理研究所 | Digital PCR device and single-pump liquid drop generating system thereof |
| CN111804353A (en) * | 2020-07-14 | 2020-10-23 | 浙江大学 | Device and method for realizing micro-droplet passive fusion |
| CN112852706A (en) * | 2019-11-28 | 2021-05-28 | 中国科学院大连化学物理研究所 | 3D (three-dimensional) organ engineering method based on aqueous two-phase droplet microfluidics |
| CN112844501A (en) * | 2019-11-28 | 2021-05-28 | 中国科学院大连化学物理研究所 | Multi-liquid-core hydrogel microcapsule chip based on double aqueous phases and application thereof |
| CN113209359A (en) * | 2021-04-26 | 2021-08-06 | 青岛大学 | Alkylated chitosan hemostatic microcapsule and preparation method thereof |
| CN113304790A (en) * | 2021-05-26 | 2021-08-27 | 南京工业大学 | Three-dimensional microfluidic chip for realizing high-throughput preparation of micro-droplets by parallelization design |
| CN113477284A (en) * | 2021-06-18 | 2021-10-08 | 上海市宝山区吴淞中心医院 | Three-dimensional cross-type liquid drop generation micro-fluidic device |
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Cited By (9)
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|---|---|---|---|---|
| CN112852706A (en) * | 2019-11-28 | 2021-05-28 | 中国科学院大连化学物理研究所 | 3D (three-dimensional) organ engineering method based on aqueous two-phase droplet microfluidics |
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| CN113304790A (en) * | 2021-05-26 | 2021-08-27 | 南京工业大学 | Three-dimensional microfluidic chip for realizing high-throughput preparation of micro-droplets by parallelization design |
| CN113477284A (en) * | 2021-06-18 | 2021-10-08 | 上海市宝山区吴淞中心医院 | Three-dimensional cross-type liquid drop generation micro-fluidic device |
| CN113477284B (en) * | 2021-06-18 | 2022-08-02 | 上海市宝山区吴淞中心医院 | Three-dimensional cross-type liquid drop generation micro-fluidic device |
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