CN108970403A - A kind of giant liposomes preparation and separating structure and method based on dielectrophoresis - Google Patents
A kind of giant liposomes preparation and separating structure and method based on dielectrophoresis Download PDFInfo
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- CN108970403A CN108970403A CN201810948022.9A CN201810948022A CN108970403A CN 108970403 A CN108970403 A CN 108970403A CN 201810948022 A CN201810948022 A CN 201810948022A CN 108970403 A CN108970403 A CN 108970403A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D57/00—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C
- B01D57/02—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C by electrophoresis
Abstract
The present invention provides a kind of giant liposomes preparation and separating structure and method based on dielectrophoresis, the structure includes the insulating layer of basal electrode layer and surface disposed thereon, basal electrode layer is equipped with interconnected preparation structure, separating structure, preparation structure includes preparing chamber and positioned at the upper electrode prepared above chamber in insulating layer, separating structure includes the sorting microchannel for sorting liposome, it prepares chamber and is connected to sorting microchannel by transition passage, sorting microchannel is located in insulating layer, basal electrode layer is equipped with the sorting electrode for being located at sorting microchannel side.The present invention applies electric signal by electrode and piezoelectric ceramic wafer is combined to issue ultrasonic signal, efficient preparation giant liposomes, it prepares reproducible, the giant liposomes of different-grain diameter are sorted by dielectrophoresis, preparation efficiency and the efficiency of separation are significantly improved, and non-breakable, sorting obtains giant liposomes similar in more partial size.
Description
Technical field
The present invention relates to liposome preparation and separating structure fields, more particularly to a kind of huge rouge based on dielectrophoresis
Plastid preparation and separating structure and method.
Background technique
Liposome is made of one layer or multilayer lipid bilayer, and inside is the closure vesica of water phase.It has double
Parent's property, has the characteristics such as class membrane structure at biocompatibility, has been applied to medicine, beauty, food and biochemistry in recent years
Etc. multiple research fields.According to the particle size of liposome, liposome can be divided into: small-sized liposome (0.02 μm of -0.2 μ of diameter
M), large-scale liposome (0.2 μm -1 μm of diameter), giant liposomes (diameter > 1 μm).Wherein, giant liposomes not only have aforementioned
Liposome common intrinsic, have more some unique properties, such as: have micron order size, be easily manipulated and in optical microphotograph
It is easy to observe under mirror, size is up to cell size, membrane structure similar cell film etc..Thus, giant liposomes have more wide
Application, be increasingly valued by people.It can encapsulate macromolecular drug, the objects such as transporter matter, DNA, plasmid, microballoon
Matter is transmitted for gene transfer, drug；It can be used as cell model, simulate cellular environment, carry out protein expression or cell function
The research of energy etc.；Also it can be used as membrane modle to study its physical property, such as mechanical performance and electrical properties；It can be also used for
Study the process and mechanism of interaction and the film perforation, fusion of molecule and film；Also it can be used as biochemical reactor, life is provided
Change the extra small reaction volume of reaction, observes and wrap up the quick biochemical reaction in volume at it.
But giant liposomes, while bring more application values, due to its enormous size, stability is poor, molding is difficult,
Also more difficulty is brought for its preparation and subsequent application.The giant liposomes preparation method proposed at present mainly includes mild water
Legal, solvent evaporated method, micro-fluidic injection method and electric forming method.
Mild hydration method: lipid is dissolved in organic by the improvement to " film dispersion method " in traditional liposomal preparation
Phase forms lipid soln；Take appropriate lipid soln in container, evaporating completely forms lipid film after falling organic solvent；Addition
Water phase is hydrated, and avoids all external disturbances as far as possible in entire hydro-combination process, after certain time, is formed in container huge
Type liposome.
Solvent evaporated method: lipid is dissolved in organic by the improvement to " two-phase dispersion method " in traditional liposomal preparation
Phase forms lipid soln, then adds water phase, comes into full contact with organic phase with water phase, then, depressurizes organic solvent evaporation,
In evaporation process, matrix material forms giant liposomes in water phase.
Micro-fluidic injection method: this method is improved by traditional solvent injection method, and the process employs propose in recent years
Micro-fluidic chip, 2003 by Tan (Yung-Chi eh Tan, Kenneth Longmuir, Abraham P.Lee,
Microfluidic liposome generation from monodisperse droplet emulsion-towards
The realization of artificial cells.2003, summer bioengineering conference.) it mentions
Out.Firstly, water phase injection is dissolved in the specific organic solvent of lipid, by chip structure by particle shape at the oil of certain partial size
Packet water drop；Then, Water-In-Oil drop is taken out and carries out various subsequent processings, obtain giant liposomes, such as: can by being injected into
In liquid phase to remove organic solvent, organic solvent is removed, so that lipid is separated, recombination forms giant liposomes (Tan
YC,Hettiarachchi K,Siu M,et al.Controlled microfluidic encapsulation of
cells,proteins,and microbeads in lipid vesicles.Journal of the American
Chemical Society,2006,128:5656-5658.)；Or by cooling to -10 DEG C, so that drop again will after freezing
Droplet surface replaces with the lipid mixture to form lipid bilayer；And then organic solvent is removed by rotary evaporation, then
Buffer is added and is used for lipid hydration, ultimately forms giant liposomes (Sugiura S, Kuroiwa T, Kagota T, et
al.Novel method for obtaining homogeneous giant vesicles from a monodisperse
water-in-oil emulsion prepared with a microfluidic device.Langmuir,2008,24:
4581-4588.)。
Electric forming method: Angelova in 1986 et al., which is put forward for the first time, applies electric field preparation giant liposomes using electrode
(Angelova MI,Dimitrov DS.Liposome electroformation.Faraday Discussions,1986,
81:303-311.), in early days using parallel platinum wire as electrode, lipid soln is added dropwise on electrode, re-evaporation forms rouge
Plasma membrane while adding water phase, by applying electric field, prepares giant liposomes.Then, electrode gradually improves, and has also been proposed and adopts
With ito glass plane electrode, silicon microwell array electrode etc..
But there are problems in terms of preparation and sorting for existing method, such as: giant liposomes particle diameter distribution is wide, difference
Greatly, prepare poor repeatability, preparation efficiency is low, encapsulating substance encapsulation rate is low, there are organic solvent residual etc., giant liposomes systems
It needs to be post-processed again after standby, especially the giant liposomes of encapsulating substance, need giant liposomes and unencapsulated
Substance separates, and using the difficulty for increasing giant liposomes application of Conventional nano liposome, increases the throwing of human and material resources
Enter, and existing post processing mode, often using centrifugation or membrane filtration, the separating effect that there are giant liposomes is poor, easy
The problems such as giant liposomes uniform particle diameter that deformation, easily broken, sorting obtain is poor.These problems all seriously hinder huge rouge
The application of plastid.Although these preparation methods are being continuously improved, up to the present, there is various lack in all preparation methods
It falls into, the above problem can not be overcome.
Summary of the invention
In view of the foregoing deficiencies of prior art, the purpose of the present invention is to provide a kind of lipids based on dielectrophoresis
Body preparation and separating structure and method, for solving the preparation of liposome in the prior art, sorting impacts liposome, divides
The problems such as selecting efficiency lower.
In order to achieve the above objects and other related objects, first aspect present invention provides a kind of lipid based on dielectrophoresis
Body preparation and separating structure, the insulating layer including basal electrode layer and surface disposed thereon, the basal electrode layer are equipped with
Interconnected preparation structure, separating structure, the preparation structure include preparing chamber and position in the insulating layer
In preparing the upper electrode above chamber, the separating structure includes the sorting microchannel for sorting liposome, the preparation
Chamber is connected to the sorting microchannel by transition passage, and the sorting microchannel is located in the insulating layer, the substrate
Electrode layer includes the sorting electrode positioned at the sorting microchannel side.
In some embodiments of the invention, the preparation structure further includes being located under basal electrode layer lower surface
Layer piezoelectric ceramic wafer, positioned at the upper electrode prepared above chamber, the basal electrode layer includes being located at the preparation
The lower electrode of cavity bottom.
In some embodiments of the invention, the upper electrode, lower electrode are electrically connected to the first signal generator,
The basal electrode layer further includes substrate glasses, and the lower electrode is located at the upper surface of the substrate glasses, the upper electrode
Upper surface be equipped with upper layer glass, substrate glasses are as lower support structure, through preparation structure and the bottom of separating structure.
Lower electrode, upper electrode, sort electrode material be specifically as follows ITO.
In some embodiments of the invention, lower layer's piezoelectric ceramic wafer is electrically connected to first by control circuit
Signal generator.
In some embodiments of the invention, the transition passage is inclined to set, and lower end, which is connected to, described prepares chamber
Room bottom, the upper end are connected to the separating structure.
In some embodiments of the invention, the transition passage is equipped with the backgauge part that can be inserted freely into or take out.
In some embodiments of the invention, the chamber for preparing has feed pathway, and the feed pathway is communicated with
Syringe pump.
In some embodiments of the invention, the separating structure further includes sorting sample introduction chamber, the transition passage
Upper end is connected to the sorting sample introduction chamber.Sorting sample introduction chamber specifically can be set to the shapes such as round, rectangular, with accumulate compared with
More fluids is carried out continuously subsequent sorting, will not be interrupted.
In some embodiments of the invention, the end of the sorting microchannel is set there are three outlet, is respectively communicated to use
The second collection chamber, the first collection chamber for collecting remaining substance, third collection chamber of the liposome needed for collecting.
In some embodiments of the invention, further include fixed frame for fixing the preparation structure, separating structure.
Second aspect of the present invention provides a kind of liposome preparation and method for separating based on dielectrophoresis, including walks as follows
It is rapid:
1) preparation of liposome: to prepare in chamber inject needed for feed liquid, after being evaporated organic solvent, inject deionized water or
Required buffer liquid applies electric signal to chamber is prepared, after lipid film is expanded to the liposome to form required diameter range, stops applying
Power up signal applies ultrasonic signal to chamber is prepared by piezoelectric ceramic wafer, lipid film is promoted to close to form liposome；
2) sorting of liposome: new deionized water or required buffer liquid are injected in chamber to preparing, passes through control circuit
Resonant electrical signals are applied to lower layer's piezoelectric ceramic wafer, change the dielectric constant of solution inside and outside liposome, while making liposome
Into sorting microchannel, electric signal is applied to sorting electrode, sorts to obtain required liposome, required liposome by dielectrophoresis
The giant liposomes that can be required partial size are also possible to contain the liposome of content, and the content contained can be micro-
Ball, nucleic acid, albumen etc..
As described above, a kind of liposome preparation and separating structure and method based on dielectrophoresis of the invention, have with
The utility model has the advantages that the present invention passes through, electrode applies electric signal and combination piezoelectric ceramic wafer issues ultrasonic signal down, efficiently prepares rouge
Plastid, prepares reproducible, the liposome of different-grain diameter is sorted by dielectrophoresis, or contain the liposome and not of content
The content contained, preparation efficiency and the efficiency of separation are significantly improved, non-breakable, and sorting obtains similar in more partial size
Liposome.
Detailed description of the invention
Fig. 1 is shown as the liposome preparation and separating structure overall structure diagram of the embodiment of the present invention.
Fig. 2 is shown as the liposome preparation of the embodiment of the present invention and the horizontal cross-sectional structural schematic diagram of separating structure.
Fig. 3 is shown as the liposome preparation of the embodiment of the present invention and the vertical schematic cross-sectional view of separating structure.
Fig. 4 is shown as in the embodiment of the present invention figure of taking pictures of Liposomal suspensions made from preparation structure under the microscope.
Fig. 5, Fig. 6 are shown as the figure of taking pictures of Liposomal suspensions under the microscope in sorting microchannel.
Fig. 7 is shown as the second collection chamber is collected into the embodiment of the present invention giant liposomes taking pictures under the microscope
Figure.
Piece mark explanation
1-the first signal generator
2-second signal generators
3-chips
4-syringe pumps
5-preparation structures
6-separating structures
7-the first collection chamber
8-the second collection chamber
9-third collection chamber
10-control circuits
11-basal electrode layers
12-feed pathways
13-prepare chamber
14-upper electrodes
15-lower layer's piezoelectric ceramic wafers
16-transition passages
17-sorting electrodes
18-sorting microchannels
19-the first microchannel
20-the second microchannel
21-third microchannels
22-backgauge parts
23-sorting sample introduction chambers
24-fixed frames
25-insulating layers
26-lower electrodes
27-substrate glasses
28-upper layer glass
Specific embodiment
Illustrate embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this specification
Other advantages and efficacy of the present invention can be easily understood for disclosed content.The present invention can also pass through in addition different specific realities
The mode of applying is embodied or practiced, the various details in this specification can also based on different viewpoints and application, without departing from
Various modifications or alterations are carried out under spirit of the invention.
It should be noted that the basic conception that only the invention is illustrated in a schematic way is illustrated provided in the present embodiment,
Then only shown in schema with it is of the invention in related component rather than component count, shape and size when according to actual implementation draw
System, when actual implementation kenel, quantity and the ratio of each component can arbitrarily change for one kind, and its assembly layout kenel can also
It can be increasingly complex.This specification structure depicted in this specification institute accompanying drawings, ratio, size etc., only to cooperate disclosed in specification
Content be not intended to limit the invention enforceable qualifications, therefore not so that those skilled in the art understands and reads
Has technical essential meaning, the modification of any structure, the change of proportionate relationship or the adjustment of size are not influencing institute of the present invention
Under the effect of capable of generating and the purpose that can reach, it should all still fall in disclosed technology contents and obtain the range that can cover
It is interior.Meanwhile cited such as "upper" in this specification, "lower", "left", "right", " centre " and " one " term, also only just
In being illustrated for narration, rather than to limit the scope of the invention, relativeness is altered or modified, without substantive change
Under more technology contents, when being also considered as the enforceable scope of the present invention.
Fig. 1 is shown as the liposome preparation and separating structure overall structure diagram of the embodiment of the present invention, and Fig. 2 is shown as this
The liposome preparation of inventive embodiments and the horizontal cross-sectional structural schematic diagram of separating structure, Fig. 3 are shown as the embodiment of the present invention
The vertical schematic cross-sectional view of liposome preparation and separating structure.The present invention provides a kind of huge lipid based on dielectrophoresis
Body preparation and separating structure are specially a chip 3, and the chip 3 is including basal electrode layer 11 and is disposed thereon surface
Insulating layer 25, basal electrode layer 11 are equipped with interconnected preparation structure 5, separating structure 6, and preparation structure includes being located at insulation
In layer 25 prepare chamber 13 and positioned at the upper electrode 14 prepared chamber 13 above, the upper surface of upper electrode 14 is equipped with upper
Layer glass 28, separating structure 6 includes the sorting microchannel 18 for sorting liposome, prepares chamber 13 and sorting microchannel 18 is logical
The connection of transition passage 16 is crossed, sorting microchannel 18 is located in insulating layer, and basal electrode layer 11, which is equipped with, is located at sorting microchannel 18
Unilateral sorting electrode 17, positive and negative electrode are staggered unilateral in sorting microchannel 18 along fluid flow direction.Sort electrode 17
Arrangement there are many, also can according to need be arranged in sorting microchannel 18 two sides.
Preparation structure 5 further includes lower layer's piezoelectric ceramic wafer 15 positioned at 11 lower surface of basal electrode layer, lower layer's piezoelectricity pottery
Porcelain chip 15 is electrically connected to the first signal generator 1 by control circuit 10, and basal electrode layer 11 is by substrate glasses 27 and its
The lower electrode layer 26 of upper covering is constituted, and the middle part for preparing 13 side of chamber is connected to a feed pathway 12, and feed pathway 12 is communicated with
Syringe pump 4, the other side for preparing chamber 13 are communicated with a transition passage 16, and the side of the transition passage 16, which is connected to, prepares chamber
The other side of the bottom of room 13, the transition passage 16 is equipped with a U-shaped opening, and U-shaped opening facilitates Liposomal suspensions and is flowed into the right side
The sorting sample introduction chamber 23 of side, sorting sample introduction chamber 23 keep it continuous in sorting microchannel 18 for accumulating Liposomal suspensions
Flowing, U-shaped opening face sort sample introduction chamber 23, and transition passage 16 is equipped with the backgauge part 22 that can be inserted freely into or take out, no
When needing feed liquid by the transition passage 16, backgauge part 22 is inserted into transition passage 16, is played the role of that feed liquid is stopped to pass through, is kept off
Materials and parts 22 are specifically as follows trapezoidal lid, and the insertion depth of trapezoidal lid is deeper than U-shaped mouth, effectively stop feed liquid circulation.
Separating structure 6 includes that a sorting sample introduction chamber 23 makes it into point for collecting the Liposomal suspensions for preparing part
Microchannel is selected, sorting 23 side of sample introduction chamber is connected to U-shaped opening, and other side is connected to sorting microchannel 18, sorts micro- logical
The end in road 18 is set there are three outlet, and three outlets are respectively communicated to the first microchannel 19, the second microchannel 20, third microchannel
21, three outlets of sorting microchannel 18 are respectively communicated to three corresponding collection chamber, including the first collection chamber 7, second
Collection chamber 8, third collection chamber 9.
In some embodiments of the invention, the upper surface of insulating layer 25 is burnishing surface, convenient for the preparation of liposome.
In some embodiments of the invention, upper electrode 14, lower electrode layer 26 are electrically connected to the generation of the first signal
Device 1, lower layer's piezoelectric ceramic wafer 15 are electrically connected to the first signal generator 1 by control circuit 10.
It in some embodiments of the invention, further include the fixed frame 24 for being mounted on preparation structure 5,6 outside of separating structure,
Fixed frame 24 clamps basal electrode layer 11, insulating layer 25, upper electrode 14, so that feed liquid will not leak outside in chamber.
Lower layer's piezoelectric ceramic wafer 15 is installed in 24 lower part of fixed frame, and lower layer's piezoelectric ceramic wafer 15 is located at basal electrode layer 11
Bottom.
In some embodiments of the invention, transition passage 16 and the substrate for preparing chamber 13 are in 45° angle, transition passage 16
Lower end connection prepare chamber 13, upper end is connected to plane on insulating layer, 16 upper end of transition passage close to separating structure 6 so that
Liposome can smoothly enter into separating structure 6.
In some embodiments of the invention, the sorting sample introduction chamber 23 of separating structure 6 is located in insulating layer 25, bottom
Positioned at the upper surface of basal electrode layer 11.Sorting sample introduction chamber 23 specifically can be set to circle, be convenient for liquid communication.
In some embodiments of the invention, transition passage 16 is located in insulating layer 25, and sorting electrode 17 is located at substrate electricity
In pole layer 11.
In some embodiments of the invention, three collection chamber are respectively positioned in insulating layer 25, and cavity bottom is located at substrate
11 upper surface of electrode layer.
Operational process of the invention is as follows: starting the cleaning processing to the chip 3 for preparing and sorting giant liposomes, plasma
Lecithin, is then dissolved in chloroform solvent by cleaning, addition fluorescent dye DiI (1,1 '-Dioctadecyl-3,3,
3 ', 3 '-tetramethylindocarbocyanine perchlorate), after organic phase is carried out dyeing processing, it is added with
Then machine blows 5min with nitrogen, then will prepare, sort huge rouge mutually on the base plane for preparing chamber 13 of preparation structure 5
The chip 3 of plastid is placed in vacuum environment lower 2 hours, allows organic solvent evaporating completely, lipid film is formed on base plane, so
It is prepared in chamber 13 backward and deionized water (or the buffer needed) is injected through channel 12 by syringe pump 4, then use first
Signal generator 1 prepares chamber 13 by upper electrode 14,11 pairs of basal electrode layer and applies electric signal 2Vpp, 10Hz, and it is small to continue 4
When, when being expanded to form required diameter range (10-50 μm) giant liposomes to more lipid film, stop applying electrode
Electric signal, then lower layer's piezoelectric ceramic wafer 15 is applied using the first signal generator 1 and control circuit connected to it 10
Resonant electrical signals generate ultrasonic signal and act on and prepares chamber 13, after promoting lipid film to close and to form giant liposomes, close the
One signal generator 1.The dielectrophoresis of giant liposomes is influenced by its partial size and inside and outside Dielectric constant, passes through note
It penetrates pump 4 and adds dielectrophoresis buffer (or other desired buffer) to chamber 13 is prepared, while being sent out using the first signal
Raw device 1 and control circuit connected to it 10 apply resonant electrical signals to lower layer's piezoelectric ceramic wafer 15, and promotion prepares chamber
Interior feed liquid is uniformly mixed, and changes the dielectric constant of solution inside and outside liposome.
The solution prepared in chamber 13 enters separating structure 6 by transition passage 16, right by second signal generator 2
Sort electrode 17 apply electric signal, 5-10Vpp, 100KHz so that giant liposomes with much smaller than 10 μm non-giant liposomes,
Lipid powder, non-encapsulating substance etc. are separated by dielectrophoresis.Finally, required giant liposomes pass through the second microchannel
20 enter the second collection chamber 8；The substances such as remaining non-giant liposomes then pass through the first microchannel 19, third microchannel 21 and enter
First collection chamber 7, third collection chamber 9 complete sorting and collect.
Fig. 4 show the figure of taking pictures that Liposomal suspensions made from preparation structure 5 amplify 200 times under the microscope.It can from Fig. 4
To find out, giant liposomes and non-giant liposomes, lipid powder, non-encapsulating substance etc. are mixed in together, if without dividing
Choosing will be unable to exclusive PCR when progress follow-up study such as constructs cell model, miniature biochemical reactor.
Fig. 5, Fig. 6 show the figure of taking pictures of Liposomal suspensions under the microscope in sorting microchannel, can from figure
Out, the lesser liposome of partial size converges on electrode, and the biggish liposome of partial size flows forward along among channel.
Fig. 7 show the giant liposomes that intermediate collection chamber is collected into, comparison diagram 7 and Fig. 4 it can be found that huge lipid
Body is effectively collected, rather than giant liposomes, lipid powder, non-encapsulating substance etc. are efficiently separated.
To sum up, the present invention applies electric signal by electrode and piezoelectric ceramic wafer is combined to issue ultrasonic signal, efficiently prepares
Giant liposomes, prepare reproducible, and the giant liposomes of different-grain diameter are sorted by dielectrophoresis, or will contain content
Giant liposomes separated with the content of unentrapped, preparation efficiency and the efficiency of separation are significantly improved, non-breakable, real
Now by giant liposomes and non-giant liposomes, do not encapsulate content and efficiently separate.
The above-described embodiments merely illustrate the principles and effects of the present invention, and is not intended to limit the present invention.It is any ripe
The personage for knowing this technology all without departing from the spirit and scope of the present invention, carries out modifications and changes to above-described embodiment.Cause
This, institute is complete without departing from the spirit and technical ideas disclosed in the present invention by those of ordinary skill in the art such as
At all equivalent modifications or change, should be covered by the claims of the present invention.
Claims (10)
1. a kind of giant liposomes preparation and separating structure based on dielectrophoresis, it is characterised in that: including basal electrode layer
(11) and the insulating layer on surface disposed thereon (25), the basal electrode layer (11) are equipped with interconnected preparation structure
(5), separating structure (6), the preparation structure (5) they include being located at preparing chamber (13) in the insulating layer (25), and described point
Selecting structure (6) includes the sorting microchannel (18) for sorting liposome, described to prepare chamber (13) and the sorting microchannel
(18) it is connected to by transition passage (16), the sorting microchannel (18) is located in the insulating layer (25), the basal electrode
Layer (11) includes the sorting electrode (17) positioned at described sorting microchannel (18) side.
2. giant liposomes preparation according to claim 1 and separating structure, it is characterised in that: the preparation structure (5)
It further include preparing chamber (13) positioned at lower layer's piezoelectric ceramic wafer (15) of basal electrode layer (11) lower surface, positioned at described
The upper electrode (14) of top, the basal electrode layer (11) include being located at the lower electrode for preparing chamber (13) bottom
(26)。
3. giant liposomes preparation according to claim 2 and separating structure, it is characterised in that: the upper electrode
(14), lower electrode (26) is electrically connected to the first signal generator (1), and the basal electrode layer (11) further includes substrate glasses
(27), the lower electrode (26) is located at the upper surface of the substrate glasses (27), and the upper surface of the upper electrode (14) is equipped with
Upper layer glass (28).
4. giant liposomes preparation according to claim 2 and separating structure, it is characterised in that: lower layer's piezoelectric ceramics
Chip (15) is electrically connected to the first signal generator (1) by control circuit (10).
5. giant liposomes preparation according to claim 1 and separating structure, it is characterised in that: the transition passage (16)
Be inclined to set, lower end be connected to it is described prepare chamber (13) bottom, the upper end is connected to the separating structure (6).
6. giant liposomes preparation according to claim 5 and separating structure, it is characterised in that: the transition passage (16)
It is equipped with the backgauge part (21) that can be inserted freely into or take out.
7. giant liposomes preparation according to claim 1 and separating structure, it is characterised in that: described to prepare chamber (13)
It is communicated with feed pathway (12), the feed pathway (12) is communicated with syringe pump (4).
8. giant liposomes preparation according to claim 1 and separating structure, it is characterised in that: the separating structure (6)
It further include sorting sample introduction chamber (23), the upper end of the transition passage (16) is connected to the sorting sample introduction chamber (23).
9. giant liposomes preparation according to claim 1 and separating structure, it is characterised in that: further include for fixing
The fixed frame (25) of preparation structure (5), separating structure (6) is stated, the end of sorting microchannel (18) is set there are three outlet, point
It is not connected to the second collection chamber (8) for collecting required liposome, for collecting unqualified liposome or other substances
First collection chamber (7), third collection chamber (9).
10. a kind of giant liposomes preparation and method for separating based on dielectrophoresis, includes the following steps:
1) preparation of liposome: required feed liquid is injected in chamber to preparing, after being evaporated organic solvent, injects deionized water or required
Buffer applies electric signal to chamber is prepared, and after lipid film is expanded to the liposome to form required diameter range, stops applying electricity
Signal applies ultrasonic signal to chamber is prepared by piezoelectric ceramic wafer, lipid film is promoted to close to form liposome；
2) sorting of liposome: new deionized water or required buffer liquid are injected in chamber to preparing, through control circuit under
Layer piezoelectric ceramic wafer applies resonant electrical signals, changes the dielectric constant of solution inside and outside liposome, while liposome is entered
Microchannel is sorted, electric signal is applied to sorting electrode, sorts to obtain required giant liposomes by dielectrophoresis.
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| CN115228407A (en) * | 2021-04-23 | 2022-10-25 | 重庆融海超声医学工程研究中心有限公司 | Apparatus and method for producing lipid vesicle |
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| 周煜,李德胜,李浩群,刘本东: "一种压电喷墨打印机喷头的动力学特性研究", 纳米技术与精密工程 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115228407A (en) * | 2021-04-23 | 2022-10-25 | 重庆融海超声医学工程研究中心有限公司 | Apparatus and method for producing lipid vesicle |
| CN115228407B (en) * | 2021-04-23 | 2023-08-22 | 重庆融海超声医学工程研究中心有限公司 | Preparation device and preparation method of lipid vesicles |
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