CN101279232A - Preparation of microballoons based on microfluid - Google Patents
Preparation of microballoons based on microfluid Download PDFInfo
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Abstract
A microfluid-based microballoon preparation method adopts a microfluid channel system to lead a colloidal solution or a pre-polymer solution to form liquid drips in a mobile phase. The formation of the liquid drip-based template is used for bioanalysis and polymeric micro spheres of protein, gene and drug screening carrier. The preparation method includes the following steps: preparation of the microfluid channel: establishing a microfluid channel network by adopting the micro-machining technology or connecting a T-shaped channel with a pinhead, a polymer pipe and a tee joint (the channel is provided with two inlets, namely, a dispersed phase inlet and a continuous phase inlet and an outlet); preparation of the liquid drip template: encasing the two phase solutions of a dispersed phase solution and a continuous phase solution respectively into a pin case, connecting with the respective inlet, digital controlling an ejector jet pump to control the velocity of flow of the two phase solutions and acquiring even and stable liquid drip template from the outlet; preparation of the microballoon: drying and solidifying the liquid drip template and removing the impurities on the surface or inside to acquire a polymer microballoon according with the requirement of bioanalysis and protein, gene and drug screening carrier.
Description
Technical field
What the present invention relates to is a kind of method for preparing microsphere based on microfluid.This preparation method is by forming drop by continuous phase liquid to effects such as the shearing force of decentralized photo liquid, surface tension, and is dry by solidifying again, forms the polymer microballoon of carrier requirements such as meeting bioanalysis and protein, gene, drug screening.
Background technology
Along with the continuous development of biomedical technology, be that the liquid-phase chip technology of carrier has obtained increasing utilization with the microballoon in bioanalysis and protein, gene, drug screening.With respect to other forms of solid phase carrier, microballoon has significant advantage: the first, and the specific area of microballoon is big, can increase the effecting reaction surface to volume ratio, and the chemical reaction on surface is carried out in littler volume; The second, adopt microballoon can utilize some other supplementary means to realize a kind of reaction system between solid-liquid reaction and liquid-liquid reactions, thereby accelerate the reaction speed of system as stir, liquid washes away etc. as carrier; The 3rd, the molecule of microsphere surface combination can be separated from solution after reaction is finished easily; The 4th, along with the change of microsphere surface functionalization group, can expand the purposes of microballoon.
At present, preparation multi-functional, the high-performance polymer microballoon is a focus of studying both at home and abroad always, and has penetrated in the research of numerous related disciplines.Method for preparing polymer micro commonly used has emulsion polymerization, dispersion copolymerization method, seeding polymerization method, suspension polymerization and microsuspension polymerization method etc., but the microspherulite diameter of this method preparation is in nanometer and sub-micrometer range, and the difficulty that certain particle is coated in the polymer is very big.And prepare hundreds of micron a method spray drying process and template etc. are arranged, but all have problems such as the inhomogeneous and making apparatus requirement height of particle diameter, cost of manufacture height to several millimeters particle diameter polymer microballoons.Microflow control technique is meant the employing Micrometer-Nanometer Processing Technology, on more than one square centimeters chip, produce the microchannel network structure, the laboratory main equipment is integrated on the as far as possible little operating platform, in order to finishing different experimentations, and the technology that can analyze product.It not only makes the consumption of reagent reduce, and speed of experiment is improved, and expense reduces, and has demonstrated fully the development trend of current laboratory equipment microminiaturization, integrated and portability.Therefore, exploitation will be simplified experimental procedure based on the method for preparing microsphere of microflow control technique, dwindle experimental facilities, accurately control the result, realize a large amount of preparations of microballoon.
Summary of the invention
Technical problem: the purpose of this invention is to provide a kind of method for preparing microsphere based on microfluid, this method preparation cost is low, is convenient to integrated and microminiaturization, and the microballoon monodispersity of preparation is good, favorable repeatability.
Technical scheme: the present invention is by forming the drop template by continuous phase liquid to effects such as the shearing force of decentralized photo liquid, surface tension, dry by solidifying again, form the polymer microballoon of carrier requirements such as meeting bioanalysis and protein, gene, drug screening.
The method for preparing microsphere that the present invention is based on microfluid adopts the microfluidic channel system to make emulsion or preceding aggressiveness solution form drop in flowing mutually, be formed for the polymer microballoon of carriers such as bioanalysis and protein, gene, drug screening based on the drop template, concrete preparation method may further comprise the steps:
1) preparation of microfluid passage: adopt micro-processing technology to set up the microfluidic channel network, perhaps select syringe needle, polymer pipe, threeway to connect a T shape passage, this passage has 2 inlets, is respectively decentralized photo inlet and continuous phase inlet, and 1 outlet is arranged;
3) preparation of drop template: decentralized photo and continuous phase two phase liquid are respectively charged into syringe, connect inlet separately,, obtain uniform and stable drop template from outlet with digital control syringe pump control two phase liquid flow velocity;
3) preparation of microballoon: with drop template dry solidification, remove surface or inner impurity, obtain meeting the polymer microballoon of bioanalysis and protein, gene, the requirement of drug screening carrier.
Described microfluidic channel network using silicon or glass or polymethyl methacrylate or dimethyl silicone polymer micro-fluid chip are made.
Described polymer pipe is polytetrafluoroethylene (PTFE) Teflon or polyether-ether-ketone Peek or polyvinyl chloride pipe.
It is continuous phase that described drop template is selected oil phase for use, and water is that decentralized photo prepares water in oil drop template, and perhaps selecting water for use is continuous phase, and oil phase is that decentralized photo prepares oil-in-water drop template; Oil phase is selected a kind of in silicone oil, paraffin oil, toluene, the dichloro hexane, and water is selected water or alcohol.
Described colloidal solution is a kind of in silicon dioxide colloid particle solution or colloidal tio 2 particle solution or the polystyrene type polymeric colloid particle solution; Before aggressiveness solution be polyvinyls solution, or teos solution, or a kind of in positive four butyl acetate solutions of peptide acid.
Diameter of micro ball is controlled, by increasing the decentralized photo flow velocity or reducing the continuous phase flow velocity microsphere diameter is increased, and reduces the decentralized photo flow velocity or increase the continuous phase flow velocity microsphere diameter is reduced.
Beneficial effect:, utilize microfluidic channel to prepare microballoon and have the following advantages according to the present invention:
1) preparation cost is low: as long as make the preparation that a microfluid system can be realized a series of microballoons.
2) be convenient to integrated and microminiaturization: because the size of microfluidic channel at several cubic centimetres, is injected if add Micropump, so whole device will be very small.
3) the liquid requirement is little: because the microchannel is a micron order or millimetre-sized, therefore can realize the flow velocity of μ l/min, reduce the waste of liquid.
4) Zhi Bei microballoon kind is many: can prepare the microballoon of different size scope by the internal diameter that changes outlet or decentralized photo outlet syringe needle, can prepare the microballoon of different materials by using different decentralized photos.
5) Zhi Bei microballoon monodispersity is good: because drop is the active force generation by two phase liquid, and two phase liquid is all very stable in passage, so produced the drop of diameter homogeneous, has both obtained the good microballoon of monodispersity behind the dry solidification.
6) favorable repeatability: because the material behavior of the size of microballoon and device is irrelevant, so but as long as the identical just repeated experiments result of two-phase liquid with previous experiments.
7) through after the corresponding curing processing, the Stability Analysis of Structures of microballoon can satisfy the instructions for use as bio-carrier preferably.
Description of drawings
Fig. 1 is a microfluidic channel schematic diagram of the present invention, and the figure acceptance of the bid is marked with: continuous phase inlet 11, decentralized photo inlet 12, outlet 13, pin hole 14.
Fig. 2 shears the microfluidic channel schematic diagram for the present invention converges, the figure acceptance of the bid is marked with: continuous phase passage 21, decentralized photo passage 22, T shape channel interface 23, collection cups 24.
The specific embodiment
We have designed a kind of microfluidic channel system (as Fig. 1).The employing micro-processing technology is set up microfluidic channel network system (comprising silicon, glass, polymethyl methacrylate, dimethyl silicone polymer micro-fluid chip) or is selected syringe needle, polymer pipe (as Teflon, Peek, polyvinyl chloride), threeway (as Peek, stainless steel) etc. to connect a T shape channel system, there are 2 inlets in this system, be respectively decentralized photo inlet and continuous phase inlet, 1 outlet is arranged.This microfluidic channel network has finally formed monodispersed oil-in-water or water in oil drop template by effects such as the shearing force between water and the oil phase, surface tension.
Adopt the microfluidic channel system to make colloidal solution or preceding aggressiveness solution form drop in flowing mutually, be formed for the polymer microballoon of bioanalysis and protein, gene, drug screening carrier based on the drop template, specifically the preparation method may further comprise the steps:
Preparation of microfluid passage: adopt micro-processing technology to set up the microfluidic channel network, perhaps select syringe needle, polymer pipe, threeway to connect a T shape passage, this passage has 2 inlets, is respectively decentralized photo inlet 11 and continuous phase inlet 12, and 1 outlet 13 is arranged;
The preparation of drop template: decentralized photo and continuous phase two phase liquid are respectively charged into syringe, connect inlet separately,, obtain uniform and stable drop template from exporting 13 with digital control syringe pump control two phase liquid flow velocity;
The preparation of microballoon: with drop template dry solidification, remove surface or inner impurity, obtain meeting the polymer microballoon of bioanalysis and protein, gene, the requirement of drug screening carrier.
It is continuous phase that described drop template is selected oil phase for use, and water is that decentralized photo prepares water in oil drop template, and perhaps selecting water for use is continuous phase, and oil phase is that decentralized photo prepares oil-in-water drop template; Oil phase is selected a kind of in silicone oil, paraffin oil, toluene, the dichloro hexane, and water is selected water or alcohol.Described colloidal solution is a kind of in silicon dioxide colloid particle solution or colloidal tio 2 particle solution or the polystyrene type polymeric colloid particle solution; Before aggressiveness solution be polyvinyls solution, or teos solution, or a kind of in positive four butyl acetate solutions of peptide acid.Collect outlet drop template,, remove the microballoon that surface or inner impurity obtain being used for carrier through dry solidification.
Regulate the flow velocity of two-phase and the concentration of decentralized photo and just can obtain the different-diameter microballoon, use different decentralized photos just can obtain the microballoon of different materials.
The present invention is a kind of microsphere preparation technology based on microfluid, and passage has two kinds of preparation methods: the one, and adopt micro-processing technology to set up microfluidic channel system (comprising silicon, glass, polymethyl methacrylate, dimethyl silicone polymer micro-fluid chip); The 2nd, select syringe needle, polymer pipe (as Teflon, Peek, polyvinyl chloride), threeway (as Peek, stainless steel) etc. to connect a T shape channel system.The microballoon of preparation mainly contains two kinds of structures: the one, and complete high polymer micro balloons, the 2nd, by the microballoon of basic particle assembling one-tenth.
The method that microsphere diameter is increased has: 1. increase the decentralized photo flow velocity; 2. reduce the continuous phase flow velocity.
The method that microsphere diameter is reduced has: 1. reduce the decentralized photo flow velocity; 2. increase the continuous phase flow velocity.
Embodiment one: the preparation of polystyrene microsphere
1. the preparation of passage:
1.. select for use three length for the 10cm internal diameter be the Teflon pipe of 500 μ m respectively as continuous phase, the passage of decentralized photo and outlet.Wherein an end of continuous phase pipeline inserts the inlet of No. 6 syringe needles as polyvinyl alcohol water solution; One end of decentralized photo inserts the inlet of No. 6 syringe needles as polystyrene solution, and the other end inserts the outlet of the syringe needle of one 60 μ m internal diameter 150 μ m external diameters as polystyrene solution.
2.. the Peek threeway with internal diameter 500 μ m connects above three Teflon pipe, and wherein the outlet syringe needle of decentralized photo passage is positioned on the central axis of outlet, continuous phase passage and other two channel vertical.
2. the preparation of drop template:
1.. the polystyrene solid is dissolved in the mixed solvent of dichloroethanes and benzene with 1% concentration, and the mixed proportion of dichloroethanes and benzene is 1: 1; Polyvinyl alcohol (PVA) is dissolved in the middle of the 500ml ultra-pure water, forms 10%PVA solution;
2.. with PS solution, PVA solution is respectively charged into syringe, is loaded on the digital control syringe pump, and the syringe of two-phase connects the inlet of decentralized photo and continuous phase pipeline respectively;
3.. the flow velocity of regulating PS solution is 5 μ l/min, and the flow velocity of PVA solution is 100 μ l/min;
4.. when outlet forms stable drop template, collect the drop template with the collection cups that PVA solution is housed;
3. the preparation of polystyrene microsphere
1.. receive after the drop template of some, collection cups is placed Rotary Evaporators, it is 60 ℃ that temperature is set, and rotating speed is 40rpm, begins rotation;
2.. the solvent for the treatment of PS solution evaporates fully, takes out microballoon, cleans the PVA solution that goes to the surface, and 60 ℃ of oven dry can obtain the polystyrene microsphere that diameter is 50 μ m.
Embodiment two: the preparation of silicon dioxide colloid crystal microballoon
1. the preparation of passage:
1.. with three length for the 10cm internal diameter be the Teflon pipe of 500 μ m respectively as continuous phase, the passage of decentralized photo and outlet.Wherein an end of continuous phase pipeline inserts the inlet of No. 6 syringe needles as polyvinyl alcohol water solution; One end of decentralized photo inserts the inlet of No. 6 syringe needles as polystyrene solution, and the other end inserts the outlet of the syringe needle of one 110 μ m internal diameter 300 μ m external diameters as polystyrene solution.
2.. the Peek threeway with internal diameter 500 μ m connects above three Teflon pipe, and wherein the outlet syringe needle of decentralized photo passage is positioned on the central axis of outlet, continuous phase passage and other two channel vertical.
2. the preparation of drop template:
1.. the 200nm silicon dioxide granule of 0.15g is distributed in the middle of the ultra-pure water of 1ml;
2.. silicon dioxde solution and silicon oil solution are respectively charged into syringe, are loaded on the digital control syringe pump, the syringe of two-phase connects the inlet of decentralized photo and continuous phase pipeline respectively;
3.. the flow velocity of regulating silicon dioxde solution is 10 μ l/min, and the flow velocity of silicon oil solution is 50 μ l/min;
4.. when outlet forms stable drop template, collect the drop template with the collection cups that silicon oil solution is housed;
3. the preparation of colloidal crystal microballoon
1.. receive after the drop template of some, collection cups was placed 60 ℃ of baking ovens 12 hours;
2.. behind the dry formation of drop template microballoon, microballoon is washed out from silicone oil with n-hexane;
3.. resulting microballoon is placed 700 ℃ baking oven two hours, can obtain all good 300 μ m colloidal crystal microballoons of mechanical strength and spectrum.
Claims (6)
1. method for preparing microsphere based on microfluid, it is characterized in that: adopt the microfluidic channel system to make colloidal solution or preceding aggressiveness solution in flowing mutually, form drop, be formed for the polymer microballoon of bioanalysis and protein, gene, drug screening carrier based on the drop template, concrete preparation method may further comprise the steps:
1) preparation of microfluid passage: adopt micro-processing technology to set up the microfluidic channel network, perhaps select syringe needle, polymer pipe, threeway to connect a T shape passage, this passage has 2 inlets, is respectively decentralized photo inlet (11) and continuous phase inlet (12), and 1 outlet (13) is arranged;
2) preparation of drop template: decentralized photo and continuous phase two phase liquid are respectively charged into syringe, connect inlet separately,, obtain uniform and stable drop template from outlet (13) with digital control syringe pump control two phase liquid flow velocity;
3) preparation of microballoon: with drop template dry solidification, remove surface or inner impurity, obtain meeting the polymer microballoon of bioanalysis and protein, gene, the requirement of drug screening carrier.
2. the method for preparing microsphere based on microfluid according to claim 1 is characterized in that the making of described microfluidic channel network using silicon or glass or polymethyl methacrylate or dimethyl silicone polymer micro-fluid chip.
3. the method for preparing microsphere based on microfluid according to claim 1 is characterized in that described polymer pipe is polytetrafluoroethylene (PTFE) Teflon or polyether-ether-ketone Peek or polyvinyl chloride pipe.
4. the method for preparing microsphere based on microfluid according to claim 1, it is characterized in that it is continuous phase that described drop template is selected oil phase for use, water is that decentralized photo prepares water in oil drop template, and perhaps selecting water for use is continuous phase, and oil phase is that decentralized photo prepares oil-in-water drop template; Oil phase is selected a kind of in silicone oil, paraffin oil, toluene, the dichloro hexane, and water is selected water or alcohol.
5. the method for preparing microsphere based on microfluid according to claim 1 is characterized in that described colloidal solution is a kind of in silicon dioxide colloid particle solution or colloidal tio 2 particle solution or the polystyrene type polymeric colloid particle solution; Before aggressiveness solution be polyvinyls solution, or teos solution, or a kind of in positive four butyl acetate solutions of peptide acid.
6. the method for preparing microsphere based on microfluid according to claim 1, it is characterized in that diameter of micro ball is controlled, by increasing the decentralized photo flow velocity or reducing the continuous phase flow velocity microsphere diameter is increased, reduce the decentralized photo flow velocity or increase the continuous phase flow velocity microsphere diameter is reduced.
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