CN103182333B - A kind of liposomal preparation and gathering-device and method - Google Patents
A kind of liposomal preparation and gathering-device and method Download PDFInfo
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- CN103182333B CN103182333B CN201310066314.7A CN201310066314A CN103182333B CN 103182333 B CN103182333 B CN 103182333B CN 201310066314 A CN201310066314 A CN 201310066314A CN 103182333 B CN103182333 B CN 103182333B
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Abstract
The present invention discloses a kind of liposomal preparation and gathering-device and method, and described device comprises signal generator and preparation and collects chip.Preparation and collection chip comprise preparation room, collecting chamber, waste liquid chamber, described preparation room comprises electrode pair and prepares chamber, electrode pair is by conductive tape connection signal generator, described collecting chamber comprises upper strata collection chamber and underlying collection chamber, alternating floor miillpore filter between two-layer chamber, described preparation room, is connected by microchannel between collecting chamber and waste liquid chamber.Liposomal preparation and gathering-device mainly apply the method powering up and prepare liposome, and utilizing miillpore filter to carry out collection process to liposome on the same chip, the present invention compared with prior art has following technique effect: the efficient giant liposomes prepared huge blank liposome or be loaded with medicine, albumen, particulate; Liposomal preparation and collection integration reduce pilot process; Required amount of reagent is few.
Description
Technical field
The present invention relates to liposomal preparation field, particularly a kind of liposomal preparation and gathering-device and use this device to prepare and collect the method for liposome.
Background technology
Liposome is that phospholipid bilayer tunic is assembled into the inner closed vesica for aqueous phase in aqueous phase.Liposome has amphiphilic, biocompatibility, and cell membrane and organelle film have similar structures, are widely used in research fields such as beauty treatment, food, medicine and biochemistries.The particle size range of liposome is from 10 nm to 100 μm, and wherein, the liposome that particle diameter is greater than 5 μm is called giant liposomes.Giant liposomes can be used as carrier for medicine transmission, also analog cell or organelle can be come as microreactor, biochemical reaction etc. in research body, in addition, giant liposomes also can be used for the simulation of cell membrane, carries out the research of membrane structure, function and the mechanism such as perforate membrane, fusion.
The giant liposomes preparation method of prior art mainly comprises traditional liposomal preparation, micro-fluidic injection method, powers up preparation method.Wherein traditional liposomal preparation mainly comprises several as follows:
1. film dispersion method, first matrix material is dissolved in organic phase, be contained in round-bottomed flask, decompression or logical nitrogen remove organic solvent, and round-bottomed flask diapire forms lipid membrane, then, the solution that water or other contain thing to be encapsulated is joined on lipid membrane, makes film be scattered in aqueous phase by mechanical dispersion modes such as jolting, rotation or vortexs, lipid membrane imbibition, bending closing forms liposome, and namely thing to be encapsulated is wrapped in liposome while liposome is formed.The shortcoming of the method is, the formation of lipid membrane is obvious by the impact of organic removal of solvents mode and mechanical dispersion mode, and restive lipid film is uniformly distributed, organic matter easily remains, and required time is long, and particle diameter is uneven, the amount that huge particle diameter liposome is formed is few, poor repeatability.
2. two-phase dispersion method, by matrix material is dissolved in organic solvent, add water again or other contains the solution of thing to be encapsulated, organic phase is fully contacted with aqueous phase, then, reduce pressure organic solvent evaporation, in evaporation process, matrix material forms liposome in aqueous phase, and namely thing to be encapsulated is wrapped in liposome while liposome is formed.The shortcoming of the method is, required time is longer, and the composition of organic phase on aqueous phase has impact, limits the kind of thing to be encapsulated, and device controls not good to reaction speed, and liposomal particle size is uneven, and the amount that huge particle diameter liposome is formed is few, poor repeatability.
Prepare giant liposomes owing to adopting traditional liposomal preparation and there is above-mentioned more problem, therefore, Recent study personnel propose the micro-fluidic injection method utilizing chip structure to prepare giant liposomes, utilize electrode applying electric field to prepare the electric preparation method of giant liposomes in addition, these methods all improve effect prepared by giant liposomes.
Wherein, the micro-fluidic injection method utilizing chip structure to prepare giant liposomes by
yung-Chieh Tandeng people 2003 propose (
yung-Chieh Tan, Kenneth Longmuir, Abraham P. Lee.microfluidic liposome generation from monodisperse droplet emulsion-towards the realization of artificial cells [J] .2003,
summer Bioengineering Conference.),the method is: first, aqueous phase is injected the specific organic solvent being dissolved with liposome materials, is formed the Water-In-Oil drop of certain particle diameter by chip structure; Subsequently, Water-In-Oil drop is collected out from chip, extracts, centrifugal, then be resuspended in aqueous phase, finally form giant liposomes.Afterwards, researcher progressively improves the method, and now, accurately controlled by chip structure and liquid flow, whole preparation process can complete on chip, finally exports the giant liposomes formed from chip channel.But the method needs retrofit chip, liquid with precise control flows, and limits to some extent organic equal reagent, high to requirement for experiment condition, and the efficiency of preparation giant liposomes is low.
Utilize chip structure to apply electric preparation method that electric field prepares giant liposomes by Angelova and Dimitrov proposed in 1986 (
m. I. Angelova, D. S. Dimitrov. Liposome Electro formation [J]. 1986,81:303-311.)the method first matrix material is dissolved in organic solvent the earliest, then organic phase is added drop-wise on parallel wire electrode, remove organic solvent under a nitrogen again, form lipid membrane, then, the solution that water or other contain thing to be encapsulated is added on electrode, apply the signal of telecommunication, electrode forms giant liposomes.Afterwards, researcher progressively improves the method and chip used, and proposition is larger with surface, and the transparent two panels ito glass electrode slice being easy to observe substitutes parallel wire electrode, do interelectrode spacer structure with PDMS, adjust the chip structure of spacing between electrode as required; Afterwards, propose and prepare lipid film on non-conducting material, it can be used as the substrate that liposome is formed, be placed between two electrodes, substrate is formed the chip structure of giant liposomes; And, adopt silicon to make surface array electrode structure, for the preparation of the giant liposomes of specified particle diameter as base material.But adopt non-conducting material to be placed between electrode and prepare giant liposomes, more to the restriction of non-conducting material, the giant liposomes be formed on layer of non-conductive material is difficult to observe; Adopt silicon to make array electrode structure as base material, light cannot through silicon base, and preparation process cannot be observed under inverted microscope, and in addition, adopting silicon to make surface array electrode structure as base material needs specialty processing, and processing is expensive and complicated.
In sum, the existing electric preparation method that these utilize chip structure applying electric field to prepare giant liposomes all also exists some problem to be solved, the chip that early stage electric preparation method adopts is simple, easy processing, but electric preparation process cannot accurately control, the improvement chip that the later stage proposes can control preparation process more accurately, but, also exist be difficult to observe, the shortcoming such as expensive, complicated operation.In addition, all chips proposed all do not comprise giant liposomes sorting, collection part, and the sorting of liposome and collection need to increase post-processing step, increase difficulty, add input, and affect the separating effect of giant liposomes and last collection rate.
Summary of the invention
Object of the present invention aims to provide a kind of efficient, simple, less investment, be easy to the giant liposomes preparation that realizes and the device collected and using method thereof, it had both comprised the part that electric field prepares giant liposomes, comprised again and prepared giant liposomes and the method for collecting to the device of prepared giant liposomes separation and collection part with this device of use.
In order to realize foregoing invention object, the technical solution adopted in the present invention is as follows:
This device comprises signal generator and preparation and collects the chip of liposome, wherein to prepare and the chip collecting liposome comprises preparation room, collecting chamber, waste liquid chamber and microchannel, described preparation room comprises upper and lower pair of electrodes and prepares chamber with middle, electrode is connected with signal generator by conductive tape, prepare chamber to be connected with collecting chamber by microchannel, upper strata, described collecting chamber comprises upper strata collection chamber and underlying collection chamber, alternating floor miillpore filter between two-layer chamber, underlying collection chamber is connected with waste liquid chamber by lower floor microchannel.
In liposomal preparation of the present invention and gathering-device, described signal generator is for providing the signal source of multiple interchange or DC signal to electrode.
In liposomal preparation of the present invention and gathering-device, described chip comprises base plane and the wall surface material layer on it, described prepares in the wall surface material layer that chamber, collecting chamber, waste liquid chamber be based upon on same base plane jointly.
In liposomal preparation of the present invention and gathering-device, described collecting chamber, the quantity of waste liquid chamber are one or more, and collecting chamber is identical with the quantity of waste liquid chamber, is connected one to one.
In liposomal preparation of the present invention and gathering-device, in the pair of electrodes up and down of described preparation room, basal electrode adopts base plane, and upper electrode is be positioned over to prepare above chamber and by the fixing rectangular flat of the fixed structure on wall surface material layer.
In liposomal preparation of the present invention and gathering-device, described chamber of preparing is connected with upper strata collection chamber by microchannel, upper strata, and underlying collection chamber is connected with waste liquid chamber by lower floor microchannel.
In liposomal preparation of the present invention and gathering-device, described wall surface material layer is two-layer up and down, and the chamber of preparation room is in lower layer wall surface material layer, and the upper electrode fixed structure of preparation room is positioned at upper strata wall surface material layer; The underlying collection chamber of collecting chamber is positioned at the lower floor of wall surface material layer, and upper strata collection chamber and miillpore filter are positioned at the upper strata of wall surface material layer; The chamber of waste liquid chamber is positioned at upper and lower two layers of walls cover material layer.
In liposomal preparation of the present invention and gathering-device, described preparation room is connected by the microchannel, upper strata being positioned at upper strata wall surface material layer with collecting chamber, and collecting chamber is connected by the lower floor microchannel being positioned at lower layer wall surface material layer with waste liquid chamber.
In liposomal preparation of the present invention and gathering-device, described preparation room chamber shape is circular, and described upper electrode fixed structure shape is rectangle, and the center of two structures is overlapping in the vertical, circular diameter is greater than the wide of rectangle, can expose the arcuate region of both sides.
Other feature of this device is, described base plane is the smooth flat that upper surface is coated with a kind of conductive material ITO or gold.Described wall surface material layer is PDMS or silicon rubber, and one insulate and plastic material.Described flat board is the smooth surface flat board that lower surface is coated with a kind of conductive material ITO or gold.Described microporous membrane surface is smooth, and aperture is accurate, and absorption is few, and come off without medium, aperture >=5 μm, material is Merlon.
Adopt the method for above-mentioned liposomal preparation and gathering-device preparation and collection liposome, technical scheme is: prepare the organic solvent adding in chamber and be dissolved with matrix material, carry out again vacuumizing process, until organic solvent evaporation until after matrix material forms dry lipid film in chamber substrate; Be fixed on by upper electrode in the wall surface material layer of upper strata, the arcuate region exposed by preparation room chamber both sides in chamber, add water or other solution containing thing to be encapsulated prepares liposome by signal generator to the preparation room electrode applying signal of telecommunication again; After required giant liposomes to be formed, shutdown signal generator, adds water or other solution for rinsing, required liposome is flushed to each upper strata collection chamber by the microchannel of upper strata wall surface material layer in chamber; Be separated by miillpore filter, required liposome rests on upper strata collection chamber, and finally for collecting, remaining residue then enters underlying collection chamber, then flows into waste liquid chamber by microchannel, thus realizes preparation and the collection of required liposome.
This application of installation powers up the principle of preparation giant liposomes, giant liposomes preparation efficiency is improve by extra electric field, in addition, the present invention proposes to utilize the miillpore filter of different pore size to be separated prepared liposome on the same chip, collect the giant liposomes of required particle diameter, decrease unnecessary pilot process, simplify the sorting of giant liposomes, collect step, improve separative efficiency and the collection efficiency of giant liposomes.Except above-mentioned points, the present invention also has that volume is little, cost is low, simple operation and other advantages.
Accompanying drawing explanation
Fig. 1 is the composition schematic diagram of liposomal preparation according to an embodiment of the invention and gathering-device.
Fig. 2 is the composition schematic diagram of liposomal preparation according to an embodiment of the invention and gathering-device.
Fig. 3 and Fig. 3 A is the structure assembling Sum decomposition schematic diagram of the preparation room part of liposomal preparation of the present invention and gathering-device.
Fig. 4 and Fig. 4 A is the structure assembling Sum decomposition schematic diagram of the collecting chamber part of liposomal preparation of the present invention and gathering-device.
Fig. 5 and Fig. 5 A is the structure assembling Sum decomposition schematic diagram of the waste liquid chamber part of liposomal preparation of the present invention and gathering-device.
Fig. 6 and Fig. 6 A is the preparation room of liposomal preparation of the present invention and gathering-device and the structure chart of collecting chamber coupling part and generalized section.
Fig. 7 and Fig. 7 A is the collecting chamber of liposomal preparation of the present invention and gathering-device and the structure chart of waste liquid chamber coupling part and generalized section.
In Fig. 1 to Fig. 7: 1-signal generator, 2-prepares and collects the chip of liposome, 3-preparation room, 4-collecting chamber, 5-waste liquid chamber, 6-conductive tape, 7-basal electrode, 8-prepares chamber, 9-upper strata fixed electrode structure, 10-upper electrode, 11-underlying collection chamber, 12-upper strata collection chamber, 13-miillpore filter, 14-lower floor waste chamber, 15 upper strata waste chamber, microchannel, 16-upper strata, 17-lower floor microchannel.
Detailed description of the invention
Be below adopt the liposomal preparation of structure of the present invention and gathering-device prepare and collect the embodiment of the method for liposome, should be appreciated that following examples are only unrestricted for illustration of technical scheme of the present invention.
Embodiment 1:
The liposomal preparation of the present embodiment and gathering-device are as shown in Fig. 1, Fig. 3, Fig. 3 A, Fig. 4, Fig. 4 A, Fig. 5, Fig. 5 A, Fig. 6, Fig. 6 A, Fig. 7 and Fig. 7 A, device comprises: the signal generator 1 sending the signal of telecommunication, prepare and collect the chip 2 of liposome, the signal generator in the present embodiment with preparation and the chip collecting liposome be connected by conductive tape 6.Wherein prepare and collect in the chip 2 of liposome and be manufactured with preparation room 3, collecting chamber 4, waste liquid chamber 5.Chip 2 is made up of the wall surface material layer on base plane and its, and wall surface material layer is divided into again two-layer up and down.
Preparation room 3 comprises basal electrode 7 and upper electrode 10, for middle prepares chamber 8 between electrode, electrode is connected with signal generator 1 by conductive tape 6, basal electrode 7 is base plane, base plane is the smooth flat that upper surface is coated with conductive material ITO or gold, prepare chamber and be arranged in lower layer wall surface material layer, upper electrode 10 is for being positioned over the rectangular flat prepared and fix above chamber and by the fixed structure on wall surface material layer, and the smooth surface that flat board is coated with conductive material ITO or gold for lower surface is dull and stereotyped.Prepare chamber 8 to be connected with the upper strata collection chamber 11 of collecting chamber 4 by microchannel, upper strata 16.Prepare the shape of chamber 8 for circular, the shape of upper electrode 10 is rectangle, and the center of two structures is overlapping in the vertical, and circular diameter is greater than the wide of rectangle, exposes the arcuate region of both sides.
Collecting chamber comprises upper strata collection chamber 12 and underlying collection chamber 11, alternating floor miillpore filter 13 between two-layer chamber, and underlying collection chamber is connected with waste liquid chamber 5 by lower floor microchannel 17.The underlying collection chamber 11 of collecting chamber is positioned at the lower floor of wall surface material layer, and upper strata collection chamber 12 and miillpore filter 13 are positioned at the upper strata of wall surface material layer.
The chamber of waste liquid chamber 5 is arranged in upper and lower two layers of walls cover material layer, is divided into lower floor's waste chamber 14 and upper strata waste chamber 15.
By with lecithin and cholesterol with the quality of 5:1 than mixed dissolution in ether solvent, add fluorescent dye DiI (1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine perchlorate) organic phase is carried out dyeing process, then, this organic phase is added drop-wise on the basal electrode 7 of preparation room, then, adopting nitrogen to blow 5min allows ether evaporate, to prepare again and the chip 2 collecting liposome is positioned over vacuum environment lower 2 hours, ether is allowed to evaporate completely, after basal electrode is formed dry lipid film, deionized water is added to preparing in chamber of preparation room 3, again by conductive tape 6 connection signal generator 1 and upper electrode 10, basal electrode 7, the signal of telecommunication 2 hours are applied to preparing chamber, preparing after chamber 8 forms a large amount of required giant liposomes, shutdown signal generator 1, deionized water is added to preparing chamber, the solution prepared in chamber is flushed in the upper strata collection chamber 12 of collecting chamber 4 through microchannel, upper strata 16, through 10 μm, miillpore filter 13(aperture) filtering screening, non-giant liposomes, giant liposomes (particle diameter≤10 μm) and the broken end of liposome just flow to underlying collection chamber 11, and then flow to waste liquid chamber 5 through lower floor microchannel 17, satisfactory giant liposomes just stays upper strata collection chamber, finally use with collection again.
The collection rate adopting the present embodiment method giant liposomes (particle diameter >=10 μm) is 40%.
Embodiment 2:
The liposomal preparation of the present embodiment and gathering-device are as shown in Figure 1.
By with lecithin and cholesterol with the quality of 5:1 than mixed dissolution in ether solvent, add fluorescent dye DiI (1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine perchlorate) organic phase is carried out dyeing process, then, this organic phase is added drop-wise on the basal electrode 7 of preparation room, then, adopting nitrogen to blow 5min allows ether evaporate, to prepare again and the chip 2 collecting liposome is positioned over vacuum environment lower 2 hours, ether is allowed to evaporate completely, after basal electrode is formed dry lipid film, interpolation in chamber of preparing to preparation room 3 contains the solution of 200mM glucose and 20mM Nacl, again by conductive tape 6 connection signal generator 1 and upper electrode 10, basal electrode 7, the signal of telecommunication is applied 2 hours to preparing chamber 8, preparing after chamber forms a large amount of required giant liposomes, shutdown signal generator 1, the solution containing 200mM glucose and 20mM Nacl is added to preparing chamber, the solution prepared in chamber is flushed in the upper strata collection chamber 12 of collecting chamber 4 through microchannel, upper strata 16, through 10 μm, miillpore filter 13(aperture) filtering screening, non-giant liposomes, giant liposomes (particle diameter≤10 μm) and the broken end of liposome just flow to underlying collection chamber 11, and then flow to waste liquid chamber 5 through lower floor microchannel 17, satisfactory giant liposomes just stays upper strata collection chamber 12, finally use with collection again.
The collection rate adopting the present embodiment method giant liposomes (particle diameter >=10 μm) is 60%.
Embodiment 3:
The structure of the liposomal preparation that the present embodiment adopts and gathering-device as shown in Figure 2, it is identical for its basic structure and embodiment 1, difference is that the quantity of collecting chamber 4 and waste liquid chamber 5 respectively has 4, and they are connected one to one, to collect the giant liposomes of different-grain diameter.
By with lecithin and cholesterol with the quality of 5:1 than mixed dissolution in ether solvent, add fluorescent dye DiI (1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine perchlorate) organic phase is carried out dyeing process, then, this organic phase is added drop-wise on the basal electrode 7 of preparation room, then, adopting nitrogen to blow 5min allows ether evaporate, to prepare again and the chip 2 collecting liposome is positioned over vacuum environment lower 2 hours, ether is allowed to evaporate completely, after basal electrode is formed dry lipid film, to the solution containing microballoon adding 200mM glucose and 20mM Nacl of preparing in chamber of preparation room 3, (microspherulite diameter is 1 μm, concentration is 1 × 10^9/mL), again by conductive tape 6 connection signal generator 1 and upper electrode 10, basal electrode 7, the signal of telecommunication 2 hours are applied to preparing chamber, preparing after chamber 8 forms a large amount of required giant liposomes, shutdown signal generator 1, the solution containing 200mM glucose and 20mM Nacl is added to preparing chamber, the solution prepared in chamber is flushed in the upper strata collection chamber 12 of collecting chamber 4 through microchannel, upper strata 16, miillpore filter 13(aperture successively in four collecting chambers 4 is followed successively by 5 μm, 8 μm, 10 μm, 12 μm) filtering screening, non-huge chamber 11, and then flow to waste liquid chamber 5 through lower floor microchannel 17, satisfactory giant liposomes just stays upper strata collection chamber 12, finally use with collection again.
Adopt the present embodiment method giant liposomes, the collection rate of the giant liposomes (particle diameter >=5 μm, particle diameter >=8 μm, particle diameter >=10 μm, particle diameter >=12 μm) of each collecting chamber is followed successively by 80%, 60%, 60%, 60%.
Claims (7)
1. liposomal preparation and a gathering-device, is characterized in that: device comprises signal generator and preparation and collects the chip of liposome, is provided with preparation room, collecting chamber, waste liquid chamber and microchannel in wherein said chip; Described preparation room comprises upper and lower pair of electrodes and prepares chamber with middle, and electrode is connected with signal generator by conductive tape, prepares chamber and is connected with collecting chamber by microchannel; Described collecting chamber comprises upper strata collection chamber and underlying collection chamber, alternating floor miillpore filter between two-layer chamber, and upper strata collection chamber is communicated with by microchannel and prepares chamber, and underlying collection chamber is connected with waste liquid chamber by microchannel;
Described chip comprises base plane and the wall surface material layer on it, and described chamber, collecting chamber, waste liquid chamber and the microchannel of preparing is produced in wall surface material layer;
In the pair of electrodes up and down of described preparation room; basal electrode is base plane; described base plane is the smooth flat that upper surface is coated with conductive material ITO or gold; upper electrode is be positioned over the rectangular flat prepared and fix above chamber and by the fixed structure on wall surface material layer, and the smooth surface that flat board is coated with conductive material ITO or gold for lower surface is dull and stereotyped;
Described wall surface material layer is two-layer up and down, prepares chamber in lower layer wall surface material layer, and the upper electrode fixed structure of preparation room is arranged in upper strata wall surface material layer; The underlying collection chamber of collecting chamber is positioned at the lower floor of wall surface material layer, and upper strata collection chamber and miillpore filter are positioned at the upper strata of wall surface material layer; The chamber of waste liquid chamber is positioned at upper and lower two layers of walls cover material layer; The microchannel connecting preparation room and collecting chamber is arranged in upper strata wall surface material layer, and the microchannel connecting collecting chamber and waste liquid chamber is arranged in lower layer wall surface material layer.
2. liposomal preparation according to claim 1 and gathering-device, is characterized in that: described signal generator is for providing the signal source of multiple interchange or DC signal to electrode.
3. liposomal preparation according to claim 1 and gathering-device, is characterized in that: the quantity of described collecting chamber, waste liquid chamber is one or more, and collecting chamber is identical with the quantity of waste liquid chamber, is connected one to one.
4. liposomal preparation according to claim 1 and gathering-device, is characterized in that: described wall surface material layer is PDMS or silicon rubber, and one insulate and plastic material.
5. liposomal preparation according to claim 4 and gathering-device, it is characterized in that: the preparation chamber shape of described preparation room is for circular, described upper electrode fixed structure shape is rectangle, and the center of two structures is overlapping in the vertical, circular diameter is greater than the wide of rectangle, exposes the arcuate region of both sides.
6. liposomal preparation according to claim 4 and gathering-device, is characterized in that: described miillpore filter aperture >=5 μm, and material is Merlon.
7. one kind uses the preparation of device described in any one of claim 1-6 and collects the method for liposome, it is characterized in that: first preparing the organic solvent adding in chamber and be dissolved with matrix material, carry out vacuumizing process, until organic solvent evaporation until matrix material is after preparing lipid film chamber substrate being formed drying; Be fixed on by upper electrode in the wall surface material layer of upper strata, the arcuate region exposed by preparation room chamber both sides in chamber, add water or other contains the solution of thing to be encapsulated, then applies the signal of telecommunication by signal generator to preparation room electrode and prepare liposome; After required giant liposomes to be formed, shutdown signal generator, adds water or other solution for rinsing, required liposome is flushed to each upper strata collection chamber by the microchannel of upper strata wall surface material layer in chamber; Be separated by miillpore filter, required liposome rests on upper strata collection chamber, and finally for collecting, remaining residue then enters underlying collection chamber, then flows into waste liquid chamber by microchannel, thus realizes preparation and the collection of required liposome.
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| CN106413896B (en) * | 2014-04-10 | 2019-07-05 | 10X基因组学有限公司 | For encapsulating and dividing fluid means, system and method and its application of reagent |
| AU2016256979B2 (en) | 2015-05-04 | 2021-01-28 | Versantis AG | Method for preparing transmembrane pH-gradient vesicles |
| CN105138865A (en) * | 2015-10-10 | 2015-12-09 | 重庆科技学院 | Multi-parameter optimization method of giant liposome preparation process |
| CN107876113B (en) * | 2017-10-20 | 2019-10-11 | 重庆医科大学 | A kind of preparation of giant liposomes and sorting unit |
| CN113559798B (en) * | 2021-07-14 | 2022-05-31 | 中山大学 | Preparation method of giant liposome with controllable size and capable of being separated from substrate |
| CN115554951B (en) * | 2022-11-10 | 2024-04-30 | 重庆大学 | Chip and method for preparing uniform giant liposome |
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