CN102008983B - Microfluidic chip suitable for producing microcapsules - Google Patents
Microfluidic chip suitable for producing microcapsules Download PDFInfo
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- CN102008983B CN102008983B CN2010105261870A CN201010526187A CN102008983B CN 102008983 B CN102008983 B CN 102008983B CN 2010105261870 A CN2010105261870 A CN 2010105261870A CN 201010526187 A CN201010526187 A CN 201010526187A CN 102008983 B CN102008983 B CN 102008983B
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Abstract
The invention discloses a microfluidic chip suitable for producing microcapsules. The microfluidic chip comprises a microcapsule production part and oil-water two-phase separation part, wherein the oil-water two-phase separation part comprises a water-phase inlet, a water-phase inlet passage, an oil-phase outlet, an oil-phase outlet passage, a water-phase outlet and a separation channel; the microcapsule production part is connected with the water-phase outlet through the separation channel; the water-phase inlet passage and the oil-phase outlet passage are connected with the separation channel; and the width of the oil-phase outlet passage is less than the diameter of the microcapsules. A stable oil-water interface is formed on an oil-water two-phase contact surface of a micro-passage by adjusting flow speed; and the microcapsules in an oil phase enter a water phase from the oil phase under the hydrodynamic effect to finish oil-water two-phase separation of the microcapsules. The microfluidic chip has a simple structural design and realizes high controllability and operability of oil-water two-phase microcapsule separation, so favorable conditions are created for wrapping isolation and water-phase culture of microcapsule biological samples (cells, bacteria and the like).
Description
Technical field
The invention belongs to the micro-total analysis field, relate to a kind of structure of micro-fluidic chip, relate in particular to a kind of structure that is applicable to that microcapsules profit two is separated.
Background technology
The micro-fluidic chip technique functions comes from the nineties in 20th century, is applied to the analytical chemistry field at first, and it is architectural feature with the microchannel; With life science is main research object, through micro-electromechanical processing technology (MEMS), whole breadboard function is comprised that print preliminary treatment, reaction, separation, detection etc. are integrated on the microchip; Analysis speed is greatly improved, has the integrated level height, reagent consumption is few; Cost of manufacture is low; Characteristics such as analysis efficiency height have very extensive applicability and application prospect, are the emphasis of current micro-total analysis system research.
Microcapsules technology can be improved the material outward appearance and improve performance, is widely used in many industrial circles.Encapsulated through material is carried out, can improve the physical property (color, outward appearance, apparent density etc.) of packing material; Improve Stability of Substance, durability.Microcapsules can storage of pharmaceutical etc. the material of micro situation, and discharge when needed.At biological field, can cell or bacterium be encapsulated the entering microcapsules, thereby avoid bioactivator to avoid the influence of environment, improve the reactivity and the stability of encrusting substance.
Based on the microcapsules production technology of micro-fluidic chip, utilize the Channel Design of micro-meter scale, can accurately control the size of the microcapsules that produce; Produce speed and shape; Through the method that repeatedly encapsulates, can produce the microcapsules of multilayer, be with a wide range of applications.Yet utilizing micro-fluidic chip to produce in the method for microcapsules, generally adopt oil as continuous phase at present, water encapsulates the water generates microcapsules as decentralized photo through oil.The existence of oil phase can exert an influence to the secretion metabolism of biologically active in the microcapsules and cell, therefore needs after microcapsules prepare, microcapsules to be transferred to water from oil phase.Existent method has chemical method (fulling, extraction etc.), physical method (centrifugal process at present; Externally-applied magnetic field, electric field etc.), these methods exist complicated operation discontinuous; Micro-processing technology requires high, need deficiency such as modify to microcapsules, has limited the application of micro-fluidic chip.Therefore, need a kind ofly can realize the micro-fluidic chip that microcapsules profit two is separated easily, efficiently.
Summary of the invention
Problem to be solved by this invention is the deficiency to prior art, provides a kind of and can realize the micro-fluidic chip that microcapsules profit two is separated easily, efficiently.
The micro-fluidic chip that is applicable to microcapsules production of the present invention comprises that egative film, microcapsules produce part and profit two part that is separated; Said profit two is separated and partly comprises the water inlet; Water access road, oil phase outlet, oil phase exit passageway, water outlet and split tunnel; Microcapsules produce part, and outlet is connected through split tunnel with water, and the water access road is connected with split tunnel respectively with the oil phase exit passageway, and the width of oil phase exit passageway is less than the diameter of microcapsules.
The structure that microcapsules produce part can like T type passage (T-Channel), flow focus type passage (Flow-Focus Channel) etc. according to conventional way design.
The eliminating of oil for the benefit of, oil phase outlet number is no less than water inlet number.
Different according to the difference of photoresist that photoetching process adopts and chip material, the profit two part channel height that is separated can be 1um-100um, and best channel height depends on and the size of microcapsules is as the criterion with the diameter that is no more than microcapsules.
Width of channel depends on the size of photoetching process and microcapsules equally, and oil phase exit passageway width can be realized better separating effect less than the water access road.Microcapsules to produce the 50um diameter are example, oil phase exit passageway width W=15um, and the channel width of water inlet is W=50um.
During profit two was separated partly, the spacing of oil phase exit passageway and water access road had considerable influence to the stabilize oil water layer, and channel pitch is too little, when increasing little processed complex degree, and adjacency channel meeting reciprocal influence; Channel pitch is too big, because oil is different with the surface tension coefficient of water, is not easy to form stable laminar flow.As a kind of preferred, oil phase exit passageway spacing is 4 times of oil phase exit passageway width.
The water access road is connected with split tunnel with 90 ° of angles with the oil phase exit passageway.
Micro-fluidic chip material of the present invention can be traditional inorganic material, like glass, and quartz, silicon chips etc. also can be polymeric materials, such as polymethyl methacrylate (PMMA), dimethyl silicone polymer (PDMS), Merlon (PC) etc.
For a better understanding of the present invention, utilize the process and the principle introduction of micro-fluidic chip production microcapsules of the present invention following:
Form the Water-In-Oil microcapsules by conventional method; Entering profit two was separated after the stage; Because oil and water viscosity coefficient have a great difference, the interface of formation has very big disturbance easily, in the present invention; Through optimizing the flow velocity of chip structure design and adjusting water and oil phase, reach stable profit two phase stable interface.Through the water inlet oil in the main channel is pushed, make oil extrude from oil outlet gradually, and microcapsules because volume is big, can not flow out from the oil phase outlet, under hydromechanical effect, get into water, reach the purpose that microcapsules profit two is separated.In the operating process, mainly be the flow velocity of regulating water and oil phase, aqueous phase flow rate is excessive, can produce bigger fluctuation to oil phase, and aqueous phase flow rate is too small, has produced not the interface of stable water oil two phase laminar flows.The profit two that is unfavorable for microcapsules is separated.
Micro-fluidic chip of the present invention has following advantage: this device process is simple, need not machined electrode, and microcapsules such as are modified at complex steps; Cost is low, does not need expensive filming equipment or externally-applied magnetic field and electric field in the manufacturing process.Utilize the present invention can realize at an easy rate microcapsules profit two is separated.In view of the above, the present invention can be widely used in fields such as life science, pharmaceutical science and medical science.
Description of drawings:
Fig. 1: in the micro-fluidic chip of the present invention profit two be separated the part structural representation.
Fig. 2: microcapsules produce the micro-fluidic chip overall structure sketch map of part for the stream focus channel.
Fig. 3: the embodiment of the invention 1 pure oil and pure water two interfacial test that is separated.
Fig. 4: the embodiment of the invention 2 microcapsules produce part.
Fig. 5: the embodiment of the invention 2 water-oil separating parts.
Fig. 6: the embodiment of the invention 2 calcium alginate microcapsule profits two test experimental result that is separated.
Fig. 7: the embodiment of the invention 3 contains saccharomycete calcium alginate microcapsule profit two test that is separated.
Fig. 8: the be separated software simulation analysis at interface of the embodiment of the invention 4 profits two.
In the accompanying drawing: 1---the water inlet, 2---the water access road, 3---the oil phase outlet; 4---the oil phase exit passageway, 5---split tunnel, 6---the water exit passageway; 7---the water outlet, 8---oil phase one inlet, 9---decentralized photo reagent one inlet; 10---oil phase two inlets, 11---decentralized photo reagent two inlets, 12---the chip hybrid chamber.
The specific embodiment:
Embodiment 1:
Pure oil and pure water two interfacial test that is separated
1) chip design with manufacture, by structure shown in Figure 1, produce micro-fluidic chip.
Design structure through mapping software, with film printout template, the method for soft lithographic is prepared in the formpiston template that the preparation micro-fluidic chip is used on the silicon chip, and four road waters inlets (1) are arranged in this stencil design, and four road binders export (3) mutually.Dimethyl silicone polymer (PDMS) liquid prepolymer is inserted on the formpiston template of micro-fluidic chip of preparation, treat that the reaction of PDMS liquid prepolymer is solidified after, the demoulding and with the card punch punching obtains micro flow chip.As shown in Figure 2, the dimethyl silicone polymer for preparing (PDMS) polymer and slide are passed through oxygen plasma (Plasma) bonding, prepare the micro-fluidic chip of required good seal.
2) be example with pure oil and pure water interface, explain that structural design of the present invention can realize stable oil-water interfaces.
As shown in Figure 3, water access road (2) is connected with split tunnel (5) with 90 ° of angles with oil phase exit passageway (4); The number of oil phase outlet is 8 the tunnel, and the number of water inlet is 4 the tunnel, and oil phase outlet number is more than the water number that enters the mouth, and the purpose of such design is in order to guarantee that oil phase has enough passages to discharge; In Fig. 3; Oil phase exit passageway width is 30um, and the width of water access road is 150um, and oil phase exit passageway width is less than the water access road; The purpose of design is to guarantee that the water inlet has enough big flux like this, and the microcapsules oil phase can be extruded from the oil phase exit passageway; In Fig. 3, the exit passageway spacing is 4 times of oil phase exit passageway width, and the purpose of design is when keeping oil phase to be discharged smoothly like this, realizes stable oil-water interfaces.In test: feed pure water at water inlet (1), flow velocity is 600ul/h, with separate circulation passage (5) in pure oil contact after; Form oil-water interfaces; Oil phase is discharged from oil phase outlet (3) owing to receive the extruding of water, reaches the oil-water two-phase interfaces balance of stable state through rate of flow of fluid control.Can find out that from lab diagram the water flow can be suitably excessive, the part pure water will flow out from the oil phase outlet, reach the target of thorough discharge oil phase.
Calcium alginate microcapsule profit two test that is separated.
Chip design and making are as example 1.
1) microcapsules generation part is as shown in Figure 4 in the microfluidic chip structure: feed oil at oil phase one inlet (8); Feed sodium alginate at decentralized photo reagent one inlet (9); Because sodium alginate is water miscible reagent, focus on mouth at stream, under the extruding of oil phase; Can produce the sodium alginate micro gel capsule, shown in Fig. 4-(a).In like manner feed oil, feed calcium chloride, produce the calcium chloride microcapsules, shown in Fig. 4-(b) at decentralized photo reagent two inlets (11) through oil phase two inlet (10).Sodium alginate micro gel capsule and calcium chloride microcapsules mix in chip hybrid chamber (12), are cured as gelatinous calcium alginate microcapsule and are dispersed in the oil phase, like 4-(c), shown in the 4-(d).Control the speed of oil phase inlet (8) (10) oil, can accurately control the size of microcapsules, produce speed and shape.
2) microfluidic chip structure water-oil separating part is as shown in Figure 5: the drop that is coated with calcium alginate microcapsule gets into water-oil separating passage (5), and in Fig. 5-(a), the upper end is the water inlet; The lower end is the oil phase outlet, and calcium alginate is near oil-water separation interface the time, and certain little disturbance can take place oil-water interfaces; Shown in Fig. 5-(b), afterwards, the drop and the oil-water interfaces that contain calcium alginate merge; Calcium alginate gets into water from oil phase, shown in Fig. 5-(c).After accomplishing water-oil separating, calcium alginate microcapsule continues to flow forward at water, shown in Fig. 5-(d), can collect it in microcapsules water outlet (7).
3) calcium alginate microcapsule profit two result of the test that is separated is as shown in Figure 6, and at the microcapsules particle diameter distribution homogeneous that water is collected, it is good that shape keeps.
Contain saccharomycete calcium alginate microcapsule profit two test that is separated.
Chip design and making are as example 1.
1) saccharomycete is unicellular fungi, can be used for brewageing production.The saccharomyces cerevisiae that adopts in this test adopts YPD (Yeast Extract Peptone Dextrose Medium) culture medium, and prescription is 1% yeast extract (Yeast Extract), 2% peptone (Peptone), 2% glucose (Dextrose Glucose).For keeping saccharomycetic activity, in experiment a few days ago, yeast is recovered; Through four all after dates that go down to posterity, contain the nutrient solution of yeast, mix with sodium alginate; Utilize the step of embodiment 2 afterwards, carry out saccharomycetic calcium alginate and encapsulate, shown in Fig. 7-(a).
2) experimental result is shown in Fig. 7-(b): in the calcium alginate gel microcapsules that solidify, be distributed with saccharomyceticly, arrow indication among the figure is the saccharomycete of budding, and promptly a saccharomycete division growth is two saccharomycete.The calcium alginate gel microcapsules are not solid sphere, and its micro-structural by many holes is propped up and is configured to, and colloidal particle in the gel or macromolecule connect mutually under certain condition; Form the space network structure; Be full of liquid in the structure space, so can utilize calcium alginate microcapsule to carry out bacterium, fungi as decentralized medium; The dimensional culture of cell has potential biological applications prospect.
The be separated software simulation analysis at interface of profit two.
1) the present invention is based on microcapsules profit two phase disengagement methods and chip structure thereof the design feasibility in theory of micro-fluidic chip for illustration, use multiple physical field coupling software Comsol to simulate, and, optimize and improve the chip structure design according to analog result.
2) software simulation setting: in Comsol software, the chip structure figure that draws, the setting of the line parameter of going forward side by side: the port number of the oil phase exit passageway (4) of simulation is an octuple, and the port number of water access road (2) is four the tunnel.The oil phase flow velocity that split tunnel (5) is set is 60ul/h, and the flow velocity of water access road (2) is 600ul/h; The pressure that oil phase outlet and water outlet are set is zero.Paint structure chart, set parameter after, structure chart is carried out gridding and calculates; The software simulation experimental result that obtains is as shown in Figure 8; Mutually independently in the oil phase exit passageway (4), have only the first via occupied by oil in octuple, other seven road binders export by water occupied mutually.Be consistent with the experimental result of embodiment 1, proved the reasonability of this structural design.
Claims (9)
1. micro-fluidic chip that is applicable to that microcapsules are produced; Comprise that microcapsules produce part and profit two part that is separated; It is characterized in that: said profit two is separated and partly comprises water inlet (1); Water access road (2), oil phase outlet (3), oil phase exit passageway (4), water outlet (7) and split tunnel (5); Microcapsules produce part and are connected through split tunnel (5) with water outlet (7), and water access road (2) is connected with split tunnel (5) respectively with oil phase exit passageway (4), and the width of oil phase exit passageway (4) is less than the diameter of microcapsules.
2. the micro-fluidic chip that is applicable to that microcapsules are produced as claimed in claim 1, it is characterized in that: oil phase exit passageway (4) width is less than water access road (2).
According to claim 1 or claim 2 be applicable to the micro-fluidic chip that microcapsules are produced, it is characterized in that: oil phase outlet (3) number is no less than water inlet (1) number.
According to claim 1 or claim 2 be applicable to the micro-fluidic chip that microcapsules are produced, it is characterized in that: oil phase exit passageway spacing is 4 times of oil phase exit passageway (4) width.
According to claim 1 or claim 2 be applicable to the micro-fluidic chip that microcapsules are produced, it is characterized in that: water access road (2) is connected with split tunnel (5) with 90 ° of angles with oil phase exit passageway (4).
According to claim 1 or claim 2 be applicable to the micro-fluidic chip that microcapsules are produced, it is characterized in that: the profit two part channel height that is separated is 1um-100um.
According to claim 1 or claim 2 be applicable to the micro-fluidic chip that microcapsules are produced, it is characterized in that: the structure that microcapsules produce part is T type passage or stream focus type passage.
According to claim 1 or claim 2 be applicable to the micro-fluidic chip that microcapsules are produced, it is characterized in that: the micro-fluidic chip material is glass, quartz, silicon chip or polymeric material.
9. the micro-fluidic chip that is applicable to that microcapsules are produced as claimed in claim 8, it is characterized in that: described polymeric material is dimethyl silicone polymer, polymethyl methacrylate or Merlon.
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| CN102553498A (en) * | 2012-02-06 | 2012-07-11 | 东南大学 | Preparation method of giant-scale porphyrin microcapsule and application thereof |
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| CN111329066A (en) * | 2020-03-30 | 2020-06-26 | 江南大学 | Method for preparing ganoderan microcapsule by using microfluidic technology |
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