CN104324769B

CN104324769B - Generation method based on the drop of microchannel

Info

Publication number: CN104324769B
Application number: CN201410655191.5A
Authority: CN
Inventors: 杜文斌; 徐鹏; 董立兵
Original assignee: Institute of Microbiology of CAS
Current assignee: Maccura Biotechnology Co ltd
Priority date: 2014-11-17
Filing date: 2014-11-17
Publication date: 2016-06-22
Anticipated expiration: 2034-11-17
Also published as: CN104324769A

Abstract

The application provides a kind of generation method of drop based on microchannel, first provides one end to have the microchannel of opening, is full of first liquid in described microchannel；The open containers filling second liquid is provided；Described microchannel is positioned at the ullage of open containers；Described first liquid and second liquid are arbitrarily immiscible two kinds of liquid or two kinds of liquid with interfacial reaction；Then described microchannel moves downward from the ullage of described open containers, makes the openings contact of described microchannel and enters described second liquid, and described first liquid is positioned at described microchannel opening part；Then the microchannel of the described second liquid of described opening entrance is upwardly away from the motion of melt surface of described open containers, the opening making described microchannel departs from second liquid, and described in be positioned at microchannel opening part first liquid depart from microchannel, in described second liquid formed drop。This drop formation method is easy, can form the drop that size is controlled, improves precision and the efficiency of high-volume drop formation。

Description

Generation method based on the drop of microchannel

Technical field

The application relates to micro liquid and measures or distribution technique field, is specifically related to a kind of generation method of drop based on microchannel。

Background technology

Under minute yardstick, the other liquid of microlitre to micro updating is manipulated accurately, it is the important proposition of the research fields such as modern project, physics, chemistry, material science, micro Process and biological engineering, has a wide range of applications in biochemical analysis, environmental monitoring, medical science and Clinical detection and micro-nano material are synthetically prepared etc.。Traditional emulsion polymerisation process (BoveyFA, etal.Emulsionpolymerization.NewYork:Intersciencepublishe rs1955, 1-22), film emulsion process (NakashimaT, etal.Membraneemulsificationbymicroporousglass.KeyEnginee ringMater1991, 513:61-61) and spraying emulsion process (LiuY, etal.Mixinginamulti-inletvortexmixer (MIVM) forfalshnano-precipitation.ChemicalEngineeringScience, 2008, 63 (11): 2829-2842) can large batch of generation particle diameter than more uniform microlayer model, but the main application of these methods is to prepare microsphere and prepare pharmaceutical carrier, be there is bigger error in the control accuracy of droplet size, cannot using the drop microreactor as accurate quantification, it is difficult to be applied in complicated biochemical reaction system。Drop formation technology based on micro-fluidic (Microfluidics) is rapidly developed (TehSY in recent years, etal.Dropletmicrofluidics.LabonaChip2008,8 (2): 198-220), the generation of its drop is based on dispersion phase and interface unstability when continuous phase crosses in microchannel。By different microfluidic channel chip designs, it is possible to generate uniform drop, and carry out merging, react and sorting etc. operates。But, the generation of the drop on micro-fluidic chip needs to meet the condition such as specific flow velocity, oil water interfacial tension and channel configurations and channel surface modification, and the scope that droplet size regulates is also affected by the restriction of factors above。It addition, after drop generates in micro-fluidic chip passage, it is necessary to specific step and device are transferred in storage container, it is difficult to the condition of single drop is customized, the operation comparatively inconvenience such as the location of drop, extraction and analysis。

By the microchannel injection such as capillary tube or injection micro liquid, and injecting liquid into micro-hole or point sample at substrate, this is the drop formation strategy of a kind of simplicity in principle。But, in practical operation, when departing from capillary tube, there is the surface tension that drop separates with continuous liquid in pipe and the adhesive force on drop and mouth of pipe surface, make the accurate quantification of droplet size be affected in drop。Generally, prior art adopts piezoelectric ceramics, heat shock expansion, high pressure EFI and special injection or the drop mode of excitation such as ultrasonic, increase drop and depart from the kinetic energy of microchannel outlet, to overcome capillary impact (TekinE, etal.Inkjetprintingasadepositionandpatterningtoolforpoly mersandinorganicparticles.SoftMatter2008,4 (4): 703-713；MeachamJM, etal.Dropletformationandejectionfromamicromachinedultras onicdropletgenerator:Visualizationandscaling.PhysicsofFl uids2005,17 (10): 100605；FerraroP, etal.Dispensingnano-picodropletsandliquidpatterningbypyr oelectrodynamicshooting.NatureNanotechnology2010,5:429-435), the particular configuration exported by microchannel and silanization or coating are processed, reduce the drop adhesive force (TavanaH at the mouth of pipe, etal.Nanolitreliquidpatterninginaqueousenvironmentsforsp atiallydefinedreagentdeliverytomammaliancells.NatureMate rials2009,8 (9): 736-741)。

But, these modes above-mentioned depend on complex fluid drive apparatus, relatively costly, and the accuracy controlling of droplet size is relatively difficult。

Summary of the invention

In view of this, the application provides a kind of generation method of drop based on microchannel, the drop formation method that the application provides is simple, the drop that size is controlled can be formed, improve precision and the efficiency of the high-volume micro-reaction of drop and mixture control, for providing basis based on the complex biological of drop and Chemical activator。

The application provides a kind of generation method of drop based on microchannel, comprises the following steps:

A) provide one end to have the microchannel of opening, in described microchannel, be full of first liquid；

The open containers filling second liquid is provided；Described microchannel is positioned at the ullage of open containers；Described first liquid and second liquid are arbitrarily immiscible two kinds of liquid or two kinds of liquid with interfacial reaction；

B) in described step a), microchannel moves downward from the ullage of described open containers, makes the openings contact of described microchannel and enters described second liquid, and described first liquid is positioned at described microchannel opening part；

C) microchannel of the described second liquid of described step b) split shed entrance is upwardly away from the motion of melt surface of described open containers, the opening making described microchannel departs from second liquid, and described in be positioned at microchannel opening part first liquid depart from microchannel, in described second liquid formed drop。

Preferably, in described step a), described microchannel is single single capillary tube, single multicore capillary tube, array capillary, micro-fluidic single channel or micro-fluidic multichannel array。

Preferably, the openings of sizes of described microchannel is between 0.05 micron to 0.5 millimeter。

Preferably, described microchannel is the microchannel that opening part processes through low-surface-energy。

Preferably, in described step a), the other end of described microchannel is connected to fluid drives equipment, continuously or intermittently produces first liquid liquid stream。

Preferably, described fluid drives equipment includes peristaltic pump, syringe pump, pressure-driven pump, air pressure driving pump or driven by electroosmosis pump。

Preferably, in described step a), described open containers is the liquid storage tank array of single liquid storage tank, one-dimensional or two-dimensional arrangements。

Preferably, in described step a), described first liquid is aqueous solution, and described second liquid is oil-based liquid immiscible with water；

Or, described first liquid is aqueous solution, and described second liquid is waterborne liquid immiscible with water；

Or, described first liquid is mineral oil, and described second liquid is perfluorine oil immiscible with mineral oil；

Or, described first liquid is sodium alginate aqueous solution, and described second liquid is calcium chloride water, both Presence of an interface reactions。

Preferably, move downward described in described step b) and adopt manual operation, the operation of manual translation platform independently with the motion of melt surface being upwardly away from described open containers described in described step c) or automatically move the control mode of platform operation。

Preferably, move downward and be upwardly away from described in described step c) frequency of motion of melt surface of described open containers described in described step b) between 0.0001 hertz to 1000000 hertz, amplitude relative to liquid level between 1 micron to 1 centimetre。

Compared with prior art, the generation method of the drop that the application provides, based on microchannel (or being called microchannel), is full of first liquid in described microchannel and one end has opening；The application also provides for filling the open containers of second liquid, and described microchannel is positioned at the ullage of open containers, and described first liquid and second liquid do not dissolve each other or have interfacial reaction。In this application, described microchannel moves downward from the ullage of described open containers, makes the openings contact of described microchannel and enters described second liquid, and described first liquid is positioned at described microchannel opening part；The microchannel of the described opening described second liquid of entrance is upwardly away from the motion of melt surface of described open containers, the opening making described microchannel departs from second liquid, and described in be positioned at microchannel opening part first liquid depart from microchannel, in described second liquid formed drop。The application is by the motion of above-mentioned microchannel opening, utilize the micro liquid interface energy when gas liquid film converts and hydrodynamic shear, overcome surface tension and adhesive force that liquid exports in microchannel, the drop making the outflow microchannel mouth of pipe can successfully depart from microchannel, and forms the drop that size is controlled。Therefore the application can pass through continuous flow and at the reciprocating upper frequency of gas-liquid interface, quickly generates the drop of high-volume fixed volume, thus improving precision and efficiency that drop such as biological sample generates。Meanwhile, the microlayer model that the application provides generates method directly can obtain the drop of volume and controllable quantity in dispersion phase, it is to avoid the evaporative effect of microlayer model, simplifies extraction and the storing step of drop or microsphere；And the application can adopt microchannel simple, lower-cost generate drop, therefore, the application provide utilize microchannel formed microlayer model method simple。

It addition, the application generate drop diameter can by the flow rate of liquid in microchannel, liquid volume, the openings of sizes of microchannel, microchannel motion frequency and amplitude etc. control, the control and regulation of droplet size are more flexible；Can also pass through to change the component flowing out first liquid in microchannel or microchannel, in open containers, sequentially form the drop of multiple different component and volume, both may be used for realizing large batch of micro-volume high flux screening, ultramicron biochemical reaction and the detection of multi-step can also be realized, have broad application prospects。

Accompanying drawing explanation

In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, the accompanying drawing used required in embodiment or description of the prior art will be briefly described below, apparently, accompanying drawing in the following describes is only embodiments of the invention, for those of ordinary skill in the art, under the premise not paying creative work, it is also possible to obtain other accompanying drawing according to the accompanying drawing provided。

Fig. 1 is the droplet distribution schematic diagram that one embodiment of the application generates；

Fig. 2 is the droplet distribution schematic diagram that one embodiment of the application generates；

Fig. 3 is the droplet distribution schematic diagram that another embodiment of the application generates；

The operating procedure schematic diagram of the drop formation method that Fig. 4 provides for the embodiment of the present application 1；

Fig. 5 is the flow velocity time history plot of pure water in capillary tube in the embodiment of the present application 1；

Fig. 6 is the position time history plot of capillary tube mouth of pipe distance mineral oil liquid level in the embodiment of the present application 1；

The operating procedure schematic diagram of the drop formation method that Fig. 7 provides for the embodiment of the present application 5；

Fig. 8 is the flow velocity time history plot of pure water in capillary tube in the embodiment of the present application 5；

Fig. 9 is the position time history plot of capillary tube mouth of pipe distance mineral oil liquid level in the embodiment of the present application 5；

The schematic diagram of the drop formation method that Figure 10 provides for the embodiment of the present application 7；

The schematic diagram of the drop formation method that Figure 11 provides for the embodiment of the present application 8；

The schematic diagram of the drop formation method that Figure 12 provides for the embodiment of the present application 9；

The schematic diagram of the drop formation method that Figure 13 provides for the embodiment of the present application 10；

The schematic diagram of the drop formation method that Figure 14 provides for the embodiment of the present application 11；

The schematic diagram of the drop formation method that Figure 15 provides for the embodiment of the present application 12；

Figure 16 is that the drop that the embodiment of the present application 13 generates under the flow velocity of 2.4 mul/min tiles microscopical view bottom liquid storage tank；

Figure 17 be the embodiment of the present application 14 4.8 mul/min flow velocitys under the drop that generates tile bottom liquid storage tank microscopical view；

Figure 18 is that the drop that the embodiment of the present application 15 generates under the flow velocity of 6 mul/min tiles microscopical view bottom liquid storage tank；

Figure 19 is that the drop that the embodiment of the present application 16 generates under the flow velocity of 12 mul/min tiles microscopical view bottom liquid storage tank；

Figure 20 is the drop graph of a relation with flow velocity of the embodiment of the present application 13～21 generation；

The process schematic of the drop formation method that Figure 21 provides for the embodiment of the present application 22～26；

The schematic diagram of different solutions ratio in the drop formation method that Figure 22 provides for the embodiment of the present application 22～26；

Figure 23 is that the sodium alginate micro ball that the embodiment of the present application 27 generates is laid in the microscope imaging figure bottom liquid storage tank；

Figure 24 is the curve chart that the sodium alginate micro ball diameter that the embodiment of the present application 27 generates changes with sodium alginate concentration；

The schematic diagram of the drop formation method that Figure 25 provides for the embodiment of the present application 28；

Figure 26 is the microscopical view that the drop that the embodiment of the present application 28 generates tiles bottom liquid storage tank；

Figure 27 is the size comparison diagram of the drop of the embodiment of the present application 28 and comparative example 1～2 generation；

The schematic diagram of the drop formation method that Figure 28 provides for the application comparative example 1；

Figure 29 is the microscopical view that the drop that the application comparative example 1 generates tiles bottom liquid storage tank；

The schematic diagram of the drop formation method that Figure 30 provides for the application comparative example 2；

Figure 31 is the microscopical view of tiling bottom liquid storage tank of the drop that the application comparative example 2 generates。

Detailed description of the invention

Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is only a part of embodiment of the present invention, rather than whole embodiments。Based on the embodiment in the present invention, the every other embodiment that those of ordinary skill in the art obtain under not making creative work premise, broadly fall into the scope of protection of the invention。

This application provides a kind of generation method of drop based on microchannel, comprise the following steps:

The application provides a kind of method utilizing microchannel to generate designated volume (micro-) drop of simple and flexible, has precision height and efficiency advantages of higher, can be applicable to the generation of biological sample drop。

The main device that the invention relates to includes microchannel, its one end open；It is pre-filled with first liquid in described microchannel, in the process of microchannel motion, first liquid can be injected according to setting program；The embodiment of the present application also provides for filling the open containers of second liquid, and described microchannel is positioned at above open containers, and pipe outlet (i.e. pipeline tapping) is the gas-liquid interface of second liquid in open containers。

The method that the application provides is based on the microchannel with opening, described microchannel can be single single capillary tube, single multicore capillary tube, array capillary, and there is micro-fluidic single channel or the multichannel array of one or more opening, it is preferably single single capillary tube, single multicore capillary tube or array capillary, compares Microfluidic droplet generation chip simpler, lower in cost。

In this application, the openings of sizes of described microchannel one end is preferably between 0.05 micron to 0.5 millimeter, between 5 microns to 0.3 millimeters。In order to the drop making generation is more homogeneous, the application can adopt the microchannel of little external diameter to export, it is preferable that initial inside diameter is 100 microns, external diameter is 200 microns。In an embodiment of the application, capillary apertures draws point for conical configuration, and internal diameter is 30 microns, and external diameter is 50 microns。

In order to the drop making generation is more homogeneous, the surface of microchannel outlet can be carried out low-surface-energy process by the application, namely described microchannel is preferably the microchannel that opening part processes through low-surface-energy, makes the first liquid of outflow be more readily pulled from the mouth of pipe, forms drop in second liquid。Described low-surface-energy processes and can process for low surface energy coat, or silanization treatment, and the application preferably employs the perfluor silane outer wall to microchannel such as capillary tube and carries out silanization treatment；Described silanization treatment is technological means well known to those skilled in the art。

It is full of first liquid in herein described microchannel, and can given either continuously or intermittently inject first liquid。In the embodiment of the present application, microchannel injecting fluid can be inject continuously with the motion of microchannel mouth, it is also possible to is move to any particular location point at microchannel to start, according to setting flow velocity and time injection first liquid。

In this application, as preferably, the other end of described microchannel is connected to fluid drives equipment；Described fluid drives equipment is the fluid drives equipment being arbitrarily likely to produce accomplished continuously or intermittently property first liquid liquid stream。The application can use peristaltic pump, syringe pump, pressure-driven pump, air pressure to drive pump or driven by electroosmosis pump etc., it is preferable that micro-injection pump, precision is higher and can set volume injected to receiving upgrading。The connected mode of microchannel and fluid drives equipment is not particularly limited by the application, connects as passed through Teflon (Teflon) tubule, and ensures seal。

The application provides open containers, is pre-loaded with second liquid, and can accept and store the drop of the first liquid injected in microchannel in it。Described open containers is arbitrarily to store the microlitre open containers to ml volumes liquid, can be described as liquid storage tank；Each liquid storage tank can store the drop of more than one first liquid, and namely described open containers can as the separate storage container of single drop, it is also possible to store a large amount of drop。In this application, described open containers is preferably the liquid storage tank array of single liquid storage tank, one-dimensional or two-dimensional arrangements, it is more preferably standard 96 hole or 384 hole ELISA Plate or PCR plate, as the high-volume storage liquid pool of the drop of specific quantity, volume and component, and the array liquid pool of the mixing of multiple droplet constituent and reaction。In this application, the bottom of described open containers can be flat, round bottom or conical bottom。

In this application, described microchannel is positioned at the ullage of open containers, and the opening of described microchannel is towards the liquid level of open containers。Further, the first liquid in herein described microchannel does not dissolve each other with the second liquid in open containers or possesses interfacial reaction。

Described first liquid and second liquid can be arbitrarily immiscible two kinds of liquid, in one embodiment of the invention, described first liquid is aqueous solution, described second liquid is oil-based liquid immiscible with water, such as mineral oil (including n-tetradecane etc.), vegetable oil, silicone oil and perfluoro alkane wet goods, the drop of generation is aqueous solution droplets。Or, described first liquid is mineral oil, and such as the organic facies such as the tetradecane and normal hexane, described second liquid is perfluorine oil immiscible with mineral oil。Described first liquid and second liquid can be immiscible aqueous two-phase, in another embodiment of the present invention, described first liquid is aqueous solution, described second liquid is waterborne liquid immiscible with water, if first liquid is dextran solution, second liquid is Polyethylene Glycol (PEG) aqueous solution, and the drop of generation is dextran solution drop。

Described first liquid and second liquid can also be two kinds of liquid with interfacial reaction, in one embodiment of the invention, described first liquid is sodium alginate aqueous solution, described second liquid is calcium chloride water, if mass concentration is the calcium chloride water of 1%, both Presence of an interface reactions, the drop of generation is calcium alginate gel bead。The application can also pass through to change the component flowing out first liquid in microchannel or microchannel, in open containers, sequentially form the drop of multiple different component and volume, both may be used for realizing large batch of micro-volume high flux screening, ultramicron biochemical reaction and the detection of multi-step can also be realized, have broad application prospects。

The embodiment of the present application controls the gas-liquid interface motion of the second liquid that do not dissolve each other that the outlet of described microchannel loads in open containers, quickly generates first liquid microlayer model。In the embodiment of the present application, described microchannel moves towards open containers, namely moves downward from the ullage of described open containers, makes the liquid level of second liquid in the openings contact open containers of described microchannel, and enters described second liquid；Now, described first liquid is positioned at described microchannel opening part, and the first liquid of microchannel injection contacts and enters second liquid, can be wrapped up by second liquid。

The described control moved downward can be adopted manual operation, the operation of manual translation platform or automatically move the mode of platform operation by the application。It should be noted that described in the control that moves downward be the motion relative to open containers liquid level of the microchannel taphole, unrelated with the motion overall for passage。The path that the described microchannel mouth of pipe moves downward is to move closer to gas-liquid interface from distance second liquid liquid level position farther out, until the first liquid of the microchannel mouth of pipe or the mouth of pipe enters second liquid solution。It should be noted that in described motor process, touch second liquid with the first liquid of microchannel mouth and be as the criterion, and whether do not go deep into below liquid level with microchannel and be as the criterion；The application preferred vertical second liquid liquid level moves downward。

In this application, described in the frequency that moves downward preferably between 0.0001 hertz to 1000000 hertz, between 0.1 hertz to 1000 hertz, it is most preferred that be 50 hertz；Described motion relative to the range-amplitude display of liquid level preferably between 1 micron to 1 centimetre, between 5 microns to 0.5 millimeters。

After moving downward, microchannel described in the embodiment of the present application is upwardly away from the motion of melt surface of open containers, it is possible to return to initial position；Pipe outlet departs from the second liquid in open containers, now, the first liquid at microchannel mouth place, due to the impact that surface tension and shearing force are main active force, departs from microchannel mouth, it is left in second liquid, the microlayer model of second liquid parcel first liquid can be formed。

The control of the described motion of melt surface being upwardly away from described open containers can be adopted manual operation, the operation of manual translation platform or automatically move the mode of platform operation by the application。In this application, described in be upwardly away from the frequency of motion of melt surface of described open containers preferably between 0.0001 hertz to 1000000 hertz, between 0.1 hertz to 1000 hertz, it is most preferred that be 50 hertz；Described motion relative to the range-amplitude display of liquid level preferably between 1 micron to 1 centimetre, between 5 microns to 0.5 millimeters。

In the embodiment of the present application, microchannel causes the drop disengaging of the first liquid in the microchannel mouth of pipe and microchannel and the first liquid of generation away from the process that open containers moves；This motor process can make the drop of the first liquid of generation be collected in the second liquid in open containers。As preferably, the vertical second liquid liquid level of the application moves upward。

The application, by the reciprocating motion on second liquid gas-liquid interface of the microchannel opening, makes the first liquid being expelled to microchannel outlet overcome the surface tension with duct fluid flow and drop and the adhesive force of the microchannel mouth of pipe, makes drop disengaging microchannel form drop。The process that generation method is one or more drop formation of the application drop, this process can start, stop and set the number of times of repetition according to programme-control, to generate certain amount of drop。

The embodiment of the present application generates the drop of first liquid, when the proportion of first liquid and the proportion of second liquid are equal, described drop is in free dispersity in open containers, suspending drops can occur in open containers, as it is shown in figure 1, Fig. 1 is the droplet distribution schematic diagram that one embodiment of the application generates；When the proportion of first liquid is less than second liquid, described drop floats on the near surface of second liquid, as in figure 2 it is shown, Fig. 2 is the droplet distribution schematic diagram that one embodiment of the application generates；When the proportion of first liquid is more than second liquid, described drop can sink down into the bottom of open containers, as it is shown on figure 3, Fig. 3 is the droplet distribution schematic diagram that another embodiment of the application generates。When the embodiment of the present application adopts the open containers of conical lower portion, and the proportion of second liquid is less than first liquid simultaneously, the drop of generation can sink, and collects in cone point place。

The application generate drop diameter can by the flow rate of liquid in microchannel, liquid volume, the openings of sizes of microchannel, microchannel motion frequency and amplitude etc. control, the control and regulation of droplet size are more flexible。The embodiment of the present application generates the adjustable extent of the volume of drop can receive liter from 20 picoliters to 10, reaches three orders of magnitude。

In sum, the generation method of the drop that the application provides, based on microchannel (or being called microchannel), is full of first liquid in described microchannel and one end has opening；The application also provides for filling the open containers of second liquid, and described microchannel is positioned at the ullage of open containers, and described first liquid and second liquid do not dissolve each other or have interfacial reaction。In this application, described microchannel moves downward from the ullage of described open containers, makes the openings contact of described microchannel and enters described second liquid, and described first liquid is positioned at described microchannel opening part；The microchannel of the described opening described second liquid of entrance is upwardly away from the motion of melt surface of described open containers, the opening making described microchannel departs from second liquid, and described in be positioned at microchannel opening part first liquid depart from microchannel, in described second liquid formed drop。The application is by the motion of above-mentioned microchannel opening, utilize the micro liquid interface energy when gas liquid film converts and hydrodynamic shear, overcome surface tension and adhesive force that liquid exports in microchannel, the drop making the outflow microchannel mouth of pipe can successfully depart from microchannel, and forms the drop that size is controlled。Therefore the application can pass through continuous flow and at the reciprocating upper frequency of gas-liquid interface, quickly generates the drop of high-volume fixed volume, thus improving precision and efficiency that drop such as biological sample generates。Meanwhile, the microlayer model that the application provides generates method directly can obtain the drop of volume and controllable quantity in dispersion phase, it is to avoid the evaporative effect of microlayer model, simplifies extraction and the storing step of drop or microsphere。

In order to further illustrate the application, the generation method of a kind of drop based on microchannel the application provided below in conjunction with embodiment is specifically described, but they can not be interpreted as the restriction to the application protection domain。

Embodiment 1

The operating procedure schematic diagram of the drop formation method that Fig. 4 provides for the embodiment of the present application 1。In Fig. 4,1 is quartz capillary, and 2 is liquid storage tank, and 3 is mineral oil, and 4 is pure water drop, 5 for volume injected be 5 receive rise pure water。The internal diameter of quartz capillary 1 is 25 microns, external diameter is 50 microns, length is 5 centimetres, and quartz capillary 1 carries out silanization treatment with dichlorodimethylsilane before the use in advance so that it is the mouth of pipe is hydrophobic。By Teflon (Teflon) tubule, (internal diameter is 300 microns in the upper end of quartz capillary 1, external diameter is 600 microns) and syringe pump (HarvardApparatus, PicoElite, not shown in FIG. 4) connect, the gap epoxide-resin glue connected is filled, it is ensured that seal。On described syringe pump, one volume of outfit is the syringe (not shown in FIG. 4) of 10 microlitres。Before the use, syringe, Teflon tubule and quartz capillary 1 are full of pure water, and check liquid flow path No leakage。Liquid storage tank 2 is glass cuvette long 1 centimetre, wide 1 centimetre, high 5 centimetres, and the volume of mineral oil 3 is 4 milliliters。

When the time is 0 second, quartz capillary 1 is positioned at mineral oil 3 ullage 5 millimeters place, it is ensured that the liquid level being full of water and water in quartz capillary 1 flushes with the mouth of pipe。Quartz capillary is with the speed of 5 millimeters per second, and vertical fluid level moves downward, and inserted below mineral oil liquid level 5 millimeters 2 second moment and remains stationary；Starting syringe pump, flow velocity is 1 receive the liter/second, and pattern is volume mode, and volume injected is 5 receive liter；Through 5 seconds, close syringe pump。Now (7 second moment), below mineral oil liquid level 5 millimeters, capillary outlet defines the drop that diameter is 212 microns, and its volume is 5 receive liter。Then, capillary tube is with the speed of 5 millimeters per second, and vertical fluid level moves upward, and mentions to mineral oil liquid level 5 millimeters；When capillary tube departs from mineral oil liquid level, acting on due to surface tension etc., the drop of capillary exit departs from capillary exit, stays in mineral oil, forms pure water drop 4 (9 second moment)。Owing to the density of pure water is 1 kg/liter, the density of mineral oil is 0.85 kg/liter, and drop is than mineral oil weight, and drop sinks to bottom liquid storage tank。The application can circulate above operating procedure, generate more volume be 5 receive rise pure water drop。

In above procedure, the flow velocity of the pure water in capillary tube is over time as it is shown in figure 5, the flow velocity time history plot that Fig. 5 is pure water in capillary tube in the embodiment of the present application 1。In motor process, over time as shown in Figure 6, Fig. 6 is the position time history plot of capillary tube mouth of pipe distance mineral oil liquid level in the embodiment of the present application 1 in the position of capillary tube mouth of pipe distance mineral oil liquid level。

With direct capillary injection volume be 4 receive rise or 5 receive rise drop compared with, this method relies on the changes such as the surface tension in the contact of the liquid level of liquid in immiscible liquid storage tank and disengaging process, capillariomotor kinetic energy is made to be converted into capillary energy, so that the solution flowed out can depart from the capillary tube mouth of pipe smoothly, it is absent from liquid because the absorption problem that cannot depart from the capillary, thereby ensure that the accuracy of droplet size。Additionally, due to the use of mineral oil, the drop of generation can be prevented effectively from evaporation, it is possible to stable storage microlayer model。

Embodiment 2

Method according to embodiment 1, it is thus achieved that volume micro-2.5 receive rise drop；Being distinctive in that, the flow velocity of syringe pump is 60 receive liter/min, changes 2.5 seconds injection time into。

Embodiment 3

Method according to embodiment 1, it is thus achieved that volume micro-2.5 receive rise drop；Being distinctive in that, the flow velocity of syringe pump changes 30 into and receives liter/min, and injection time is 5 seconds。

Embodiment 4

Method according to embodiment 1, it is thus achieved that volume is the drop of 25 picoliters；Being distinctive in that, the exit inside diameter drawing point capillary tube is 8 microns, and external diameter is 15 microns, and the flow velocity of syringe pump changes 15 into and receives liter/min, and injection time is 0.1 second。

The control that above example can set according to syringe pump, the skin that accurately acquisition volume is controlled, the drop risen of receiving。Therefore, this method belongs to a kind of method quantitatively measuring and operating skin, nanoliter volumes liquid。

Embodiment 5

The operating procedure schematic diagram of the drop formation method that Fig. 7 provides for the embodiment of the present application 5。In Fig. 7,1 is quartz capillary, and 2 is liquid storage tank, and 3 is mineral oil, and 4 is pure water drop, and 5 is injection pure water。The internal diameter of quartz capillary 1 is 25 microns, external diameter is 50 microns, length is 5 centimetres, and quartz capillary 1 carries out silanization treatment with dichlorodimethylsilane before the use in advance so that it is the mouth of pipe is hydrophobic。By Teflon (Teflon) tubule, (internal diameter is 300 microns in the upper end of quartz capillary 1, external diameter is 600 microns) and syringe pump (HarvardApparatus, PicoElite, not shown in FIG. 7) connect, the gap epoxide-resin glue connected is filled, it is ensured that seal。On described syringe pump, one volume of outfit is the syringe (not shown in FIG. 7) of 10 microlitres。Before the use, syringe, Teflon tubule and quartz capillary 1 are full of pure water, and check liquid flow path No leakage。Liquid storage tank 2 is glass cuvette long 1 centimetre, wide 1 centimetre, high 5 centimetres, and the volume of mineral oil 3 is 4 milliliters。

When the time is 0 second, quartz capillary 1 is vertically placed, and initial position is mineral oil 3 ullage 5 millimeters place。Quartz capillary is with the speed of 2 mm/second, and vertical fluid level moves downward, and inserts below mineral oil liquid level 5 millimeters and changes the direction of motion of capillary tube at once, vertically up moving, be returned to initial position。Now changing the direction of motion more at once is vertically downward, enters next motion cycle。In reciprocatory movement, capillary tube connects syringe pump and also ensures to be full of in capillary tube pure water, and syringe pump injects pure water continuously with 1 flow velocity receiving the liters/second, and under above continuous injection pattern, the time of a circulation is 5 seconds, and it is 5 receive liter that capillary tube is total to volume injected。In motor process, when capillary tube departs from mineral oil liquid level, acting on due to surface tension etc., the drop of capillary exit departs from capillary exit, stays in mineral oil, forms pure water drop 4 (9 second moment)。Owing to the proportion of pure water is more than mineral oil, drop sinks to bottom liquid storage tank。Through first circulate after, follow-up circulation can be stably obtained volume be 5 receive rise pure water drop。

In operating process, over time as shown in Figure 8, Fig. 8 is the flow velocity time history plot of pure water in capillary tube in the embodiment of the present application 5 to the flow velocity of the pure water in capillary tube。In motor process, the position of capillary tube mouth of pipe distance mineral oil liquid level is over time as it is shown in figure 9, Fig. 9 is the position time history plot of capillary tube mouth of pipe distance mineral oil liquid level in the embodiment of the present application 5。

In above step, syringe pump operates without suspend and restarting etc., and the motion of capillary tube continuously performs, and is absent from suspending, and therefore the efficiency of the present embodiment generation drop is higher compared to embodiment 1, and operating procedure more simplifies。By circulate operating procedure, the application can obtain a large amount of volume be 5 receive rise pure water drop。Owing to proportion is more than mineral oil, the drop of generation sinks to bottom liquid storage tank。

Embodiment 6

Method according to embodiment 5, it is thus achieved that volume be 2.5 receive rise pure water drop。Being distinctive in that, the flow velocity of syringe pump changes 0.5 into and receives the liter/second；Mineral oil is previously added the Span80 (sorbester p17) of 5% (v/v), so that the fusion avoided droplets from when generating a large amount of drop。

Embodiment 7

The schematic diagram of the drop formation method that Figure 10 provides for the embodiment of the present application 7。In Figure 10,1 is three parallel capillary tubies, and 2 is three drops synchronizing to generate。

The parameter of the present embodiment capillary tube and the operating procedure of drop formation, with embodiment 4, are distinctive in that, the present embodiment adopts three parallel capillary tubies to synchronize to generate three drops, and wherein, intercapillary spacing is 2 millimeters。

The array capillary design of the application, it is possible to by being continuously increased the radical of capillary tube, high-volume generates drop。

Embodiment 8

The schematic diagram of the drop formation method that Figure 11 provides for the embodiment of the present application 8。In Figure 11,1 is the micro-fluidic chip containing 5 parallel drop formation passages, and 2 is 5 drops synchronizing to generate。

The operating procedure of the present embodiment drop formation, with embodiment 7, is distinctive in that, adopts micro-fluidic chip to substitute capillary tube, reaches to improve the purpose of drop formation flux equally。Wherein, micro-fluidic chip is plastic material, and passage is 30 microns of deep, 100 microns wide, 1 centimeter length。Channel pitch is 1.5 millimeters, thick 2 millimeters of chip, wide 8.5 millimeters, length 2 centimetres。Channel outlet adopts sharp-crested design, it is to avoid the drop of generation is adsorbed on chip and can not fall。

This method can simply by array microchannel, and Mass production drop, efficiency is higher。

Embodiment 9

The schematic diagram of the drop formation method that Figure 12 provides for the embodiment of the present application 9。In Figure 12,1 is twin-core capillary tube, namely has a dividing plate to be divided into two not connected passages inside a capillary, and I is first passage, and II is second channel；The 2 bi-component mixing drops for generating。

The present embodiment generates the operating procedure of drop with embodiment 1, is distinctive in that, the present embodiment utilizes twin-core capillary tube, generates the drop of bi-component mixing；Wherein, twin-core capillary tube is of a size of external diameter 300 microns, internal diameter 200 microns, thick 50 microns of central dividing plate；In twin-core capillary tube first passage I, pass into the luciferin solution of 1 mol/L, in second channel II, pass into pure water solution；The dilution drop that bi-component mixing drop is the fluorescein being mixed with both solution of described generation, the fluorescein concentration of drop is determined by the flow rate of two passages。

This method can also adopt the multicore capillary tube separated with two or more, generates the multicomponent mixing drop being mixed with two or more solution。

Embodiment 10

The schematic diagram of the drop formation method that Figure 13 provides for the embodiment of the present application 10。In Figure 13,1 is three closely side-by-side single channel capillary tubies, and I is the first capillary tube, and II is the second capillary tube, and III is three capillary；The 2 three component mixing drops for generating。

The operating procedure of the parameter of the single single channel capillary tube of the present embodiment and generation drop, with embodiment 1, is distinctive in that, the present embodiment adopts three closely side-by-side single channel capillary tubies, generates the drop of three components mixing；Wherein, potassium iodide, potassium iodate, three kinds of colourless solutions of dilute sulfuric acid it are injected separately in the first capillary tube I, the second capillary tube II and three capillary III, three component mixing drops of described generation are the drop being mixed with these three solution, owing to redox reaction generates iodine, making drop is yellowish-brown。

Embodiment 11

The schematic diagram of the drop formation method that Figure 14 provides for the embodiment of the present application 11。In Figure 14,1 confluxes chip for four-way, and I is first passage, and II is second channel, and III is third channel, and IV is fourth lane；The 2 four component mixing drops for generating。

The present embodiment generates the operating procedure of drop with embodiment 1, is distinctive in that, the present embodiment adopts four-way to conflux chip, generates the drop of four components mixing；Wherein, four-way confluxes has four passages finally to merge into a straight channel in chip, the material of chip is poly-dimethoxysilane (PDMS), is of a size of 1.5 centimetres wide, 2.5 centimetres high, 0.5 cm thick, and chip end adopts the method cut to obtain tip outlet；Being injected separately into red pigment, flavochrome, marennin, four kinds of solution of cyanine at four passages, they merge in the straight channel at rear；Four component mixing drops of described generation are the dark drop being mixed with these four pigment solution。

Embodiment 12

The schematic diagram of the drop formation method that Figure 15 provides for the embodiment of the present application 12。In Figure 15,1 is drop formation stream chip, and 2 is the emulsion phase drop generated。

The present embodiment generates the operating procedure of drop with embodiment 1, is distinctive in that, the present embodiment adopts drop formation stream chip, generates the drop of emulsion phase；Wherein, having upper and lower two entrances in drop formation stream chip, superincumbent entrance injects the mineral oil that with the addition of surfactant, divides in two passages, and the aqueous phase pinch off that porch, lower section is injected forms water in oil drop in passage in the wings；The material of chip is poly-dimethoxysilane (PDMS), be of a size of 2 centimetres wide, 3 centimeter length, 5 millimeters thick, chip end adopts blade to cut into conical design；The drop of the emulsion phase that emulsion phase drop is W/O/W generated in liquid storage tank。

The frequency of this method flow velocity and vibration by changing chip, it is possible to achieve an emulsion drop wraps up different number of drop。

Embodiment 13～16

According to the method for embodiment 1, generate drop by regulating flow velocity。It is distinctive in that, in embodiment 13～16, first liquid is deionized water, second liquid is containing 4%AbilEM90 (cetyl polyethylene/polypropylene glycol-10/1 dimethyl silica alkanol, source is for Germany/Ying Chuangaoshi Mitt USA Corporation) the tetradecane, syringe pump flow velocity respectively 2.4 mul/min, 4.8 mul/min, 6 mul/min, 12 mul/min。

Figure 16 is that the drop that the embodiment of the present application 13 generates under the flow velocity of 2.4 mul/min tiles microscopical view bottom liquid storage tank；Figure 17 be the embodiment of the present application 14 4.8 mul/min flow velocity under the drop that generates tile bottom liquid storage tank microscopical view；Figure 18 is that the drop that the embodiment of the present application 15 generates under the flow velocity of 6 mul/min tiles microscopical view bottom liquid storage tank；Figure 19 is that the drop that the embodiment of the present application 16 generates under the flow velocity of 12 mul/min tiles microscopical view bottom liquid storage tank。

The application, by measuring the radius of drop, calculates and obtains the volume of the drop of generation under different in flow rate。Result is referring to the graph of a relation of the drop that Figure 20, Figure 20 are the embodiment of the present application 13～21 generation with flow velocity。

Embodiment 17～21

According to the method for embodiment 13, generate drop by regulating flow velocity。It is distinctive in that, syringe pump flow velocity respectively 0.06 mul/min, 0.30 mul/min, 0.60 mul/min, 1.2 mul/min, 18 mul/min in embodiment 17～21。

It can be observed from fig. 20 that its corresponding flow velocity of volume of drop that this method generates under different in flow rate presents linear relationship。

Embodiment 22～26

The process schematic of the drop formation method that Figure 21 provides for the embodiment of the present application 22～26。In Figure 21,1 is 96 orifice plates with conical lower portion, and I is for receiving a liter sample drop, and II is for receiving a liter reaction reagent solution, and III is mineral oil, and I+II is the multi component droplet generated。

Embodiment 22～26 utilizes the method for embodiment 1, in each hole of 96 orifice plates 1, first generate a sample drop I receiving liter sample volume, generating another drop II containing reaction reagent again in each hole, the proportion of the two drop is bigger than oil phase, therefore sinks at the bottom of pipe；Due to V-shaped at the bottom of pipe, two drops can converge at bottom tip；When in oil phase not or containing very small amount of surfactant time, two drops just merge and become a drop I+II, and it is mixed with sample I and reaction reagent solution II。

Generating the different solutions of different volumes in different pipes, the ratio merging the different solutions that the drop formed is mixed with is different。The schematic diagram of different solutions ratio in the drop formation method provided for the embodiment of the present application 22～26 referring to Figure 22, Figure 22。In embodiment 22～26, the method utilizing embodiment 1, in five pipes, first generate respectively with sample and 10 receive liter, 20 receive liter, 30 receive liter, 40 receive liter, 50 receive the drop risen, then in this five pipe with reaction reagent solution generate respectively 50n receive liter, 40 receive liter, 30 receive liter, 20 receive liter, 10 receive liter, drop；In each pipe, two drops sink to bottom, and converge at bottom tip position and merge。

The volume of the drop of the fusion formed in each pipe is equal, is 60 and receives liter；But the sample that they are mixed with is different from the ratio of reaction reagent solution, respectively 1:5,1:2,1:1,2:1,5:1。

This method is capable of the control of large batch of multi component droplet mixed volume and ratio, can be applicable to high flux screening experiment。

Embodiment 27

According to the method for embodiment 5, by changing the composition of first liquid and second liquid, it is thus achieved that the sodium alginate micro ball of solidification。The present embodiment specifically includes:

The first liquid flowed out in capillary tube is replaced by micro-sodium alginate aqueous solution, and the second liquid in liquid storage tank is replaced by the calcium chloride water that mass concentration is 0.5%。

In use, capillary tube is vertically placed, initial position is calcium chloride water ullage 5 millimeters place, capillary tube is with the speed of 2 mm/second, vertical downward movement, inserts below calcium chloride water 5 millimeters, and changes the direction of motion of capillary tube at once, vertically up move, be returned to calcium chloride water ullage 5 millimeters place。Now changing the direction of motion more at once is vertically downward, enters next motion cycle。The time of each motion cycle is 0.02 second。In reciprocatory movement, capillary tube is full of sodium alginate aqueous solution in connecting syringe pump and ensureing capillary tube, and syringe pump is with the continuous injecting fluid of flow velocity of 0.12 mul/min。In motor process, when sodium alginate aqueous solution enters calcium chloride water by capillary tube, due to the effect of calcium ion, making sodium alginate cross-linking solidify, viscosity increases。Needing the regular hour owing to sodium alginate aqueous solution solidifies in calcium chloride water, among a circulation of capillary tube, sodium alginate aqueous solution remains to keep the state of liquid to flow out。Acting on due to surface tension etc., sodium alginate aqueous solution forms drop and departs from capillary exit and enter liquid storage tank。

Above under continuous injection pattern, difference by the capillary inner diameter used, can be stably obtained volume be 0.0042 receive rise～4.2 receive rise sodium alginate micro ball, as shown in figure 23, Figure 23 be the embodiment of the present application 27 generate sodium alginate micro ball be laid in the microscope imaging figure bottom liquid storage tank。

In order to ensure the mechanical performance of sodium alginate micro ball, after microsphere generation terminates, adding 400 microlitre mass concentrations in liquid storage tank is the calcium chloride water of 5%, makes sodium alginate micro ball solidify further。

The microsphere volume of this method is mainly determined by the internal diameter of capillary outlet, by changing the internal diameter of capillary outlet, it is possible to obtain diameter is 20 microns to the 200 microns sodium alginate micro balls not waited。

Character about the drop of this method, it is possible to changed by the concentration of sodium alginate aqueous solution and adjust。Simultaneously when sodium alginate aqueous solution concentration changes, change due to its viscosity and solidification rate, there is corresponding change in microsphere diameter, as shown in figure 24, Figure 24 is the curve chart that the sodium alginate micro ball diameter that the embodiment of the present application 27 generates changes with sodium alginate concentration。

Embodiment 28

The schematic diagram of the drop formation method that Figure 25 provides for the embodiment of the present application 28。In Figure 25,1 is capillary tube, and 2 is liquid storage tank, and 3 is the tetradecane containing 3%EM90, and 4 is pure water drop。Wherein, capillary tube 1 model is 30 micron inside diameter, 50 microns outer diameter, and capillary tube outer wall adopts dichlorodimethylsilane to carry out Silanized hydrophobic process。

According to the method for embodiment 1, generate drop。It is the microscopical view that the drop that the embodiment of the present application 28 generates tiles bottom liquid storage tank referring to Figure 26, Figure 26。

Calculating the diameter obtaining described drop, as shown in figure 27, Figure 27 is the size comparison diagram of the drop of the embodiment of the present application 28 and comparative example 1～2 generation。

Comparative example 1

According to the method for embodiment 28, generating drop, be distinctive in that, the capillary tube mouth of pipe moves back and forth under tetradecane liquid level。

Schematic diagram referring to the drop formation method that Figure 28, Figure 28 provide for the application comparative example 1。It is the microscopical view that the drop that the application comparative example 1 generates tiles bottom liquid storage tank referring to Figure 29, Figure 29。As can be seen from Figure 29, the drop of generation compares heterogeneity。Referring to Figure 27 it can be seen that the diameter of drop that this comparative example generates compares embodiment 28 fluctuates bigger。

Comparative example 2

According to the method for embodiment 28, generating drop, be distinctive in that, the capillary tube mouth of pipe moves back and forth on tetradecane liquid level。

Schematic diagram referring to the drop formation method that Figure 30, Figure 30 provide for the application comparative example 2。It is the microscopical view that the drop that the application comparative example 2 generates tiles bottom liquid storage tank referring to Figure 31, Figure 31。As can be seen from Figure 31, the drop size of generation is excessive。Referring to Figure 27 it can be seen that the diameter of drop that this comparative example generates compares embodiment 28 fluctuates bigger。

As seen from the above embodiment, the application moves at gas-liquid interface based on microchannel outlet, and injects the drop generating size controllable precise。The drop formation method that the application provides has simple, precision and efficiency relatively advantages of higher, has broad application prospects。

Claims

1., based on a generation method for the drop of microchannel, comprise the following steps:

2. generation method according to claim 1, it is characterised in that in described step a), described microchannel is single single capillary tube, single multicore capillary tube, array capillary, micro-fluidic single channel or micro-fluidic multichannel array。

3. generation method according to claim 2, it is characterised in that the openings of sizes of described microchannel is between 0.05 micron to 0.5 millimeter。

4. generation method according to claim 1, it is characterised in that described microchannel is the microchannel that opening part processes through low-surface-energy。

5. generation method according to claim 1, it is characterised in that in described step a), the other end of described microchannel is connected to fluid drives equipment, continuously or intermittently produces first liquid liquid stream。

6. generation method according to claim 5, it is characterised in that described fluid drives equipment includes peristaltic pump, syringe pump, pressure-driven pump, air pressure driving pump or driven by electroosmosis pump。

7. generation method according to claim 1, it is characterised in that in described step a), described open containers is the liquid storage tank array of single liquid storage tank, one-dimensional or two-dimensional arrangements。

8. generation method according to claim 1, it is characterised in that in described step a), described first liquid is aqueous solution, and described second liquid is oil-based liquid immiscible with water；

9. generation method according to claim 1, it is characterized in that, move downward described in described step b) and adopt manual operation, the operation of manual translation platform independently with the motion of melt surface being upwardly away from described open containers described in described step c) or automatically move the control mode of platform operation。

10. generation method according to claim 1, it is characterized in that, move downward and be upwardly away from described in described step c) frequency of motion of melt surface of described open containers described in described step b) between 0.0001 hertz to 1000000 hertz, amplitude relative to liquid level between 1 micron to 1 centimetre。