CN107059407A - It is a kind of to be used to capture nano fibrous membrane of folacin receptor height expression cancer cell and its preparation method and application - Google Patents
It is a kind of to be used to capture nano fibrous membrane of folacin receptor height expression cancer cell and its preparation method and application Download PDFInfo
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- CN107059407A CN107059407A CN201710316358.9A CN201710316358A CN107059407A CN 107059407 A CN107059407 A CN 107059407A CN 201710316358 A CN201710316358 A CN 201710316358A CN 107059407 A CN107059407 A CN 107059407A
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- nano fibrous
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- cancer cell
- fibrous membrane
- folacin receptor
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- OVBPIULPVIDEAO-LBPRGKRZSA-N Folic acid Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 title claims abstract description 67
- 239000011724 folic acid Substances 0.000 title claims abstract description 53
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- D06M15/19—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment with synthetic macromolecular compounds
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- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
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- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/54—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties by welding together the fibres, e.g. by partially melting or dissolving
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- D06M13/402—Amides imides, sulfamic acids
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Abstract
The present invention relates to a kind of nano fibrous membrane for being used to capture folacin receptor height expression cancer cell and its preparation method and application, nano fibrous membrane is the polylactic-co-glycolic acid PLGA nano fibrous membranes of modified with folic acid.Preparation method includes：(1) preparation of electrostatic spinning nano fiber film；(2) modified with folic acid of nano fibrous membrane.The nano fibrous membrane is embedded into inside micro-fluidic chip and for the specificity capture of cancer cell.Preparation technology of the present invention is simple, and high specificity can be integrated in inside micro-fluidic chip, realizes that specific efficient is captured to cancer cell, with good application prospect.
Description
Technical field
It is more particularly to a kind of to be used to capture folacin receptor height expression cancer the invention belongs to electrostatic spinning nano fiber Material Field
Nano fibrous membrane of cell and its preparation method and application.
Background technology
Research in recent years shows, the liquid biopsy based on circulating tumor cell (Circulating Tumor Cells, CTCs)
Technology is expected to turn into a kind of brand-new tumour technique for detection.Due to CTCs, quantity is considerably less in blood, and 109Individual haemocyte
In only containing 1~100 CTCs, the CTCs cells that rareness is isolated in numerous haemocytes have much difficulty, ground as CTCs
The main obstruction studied carefully.Therefore the key of CTCs researchs is from the complicated blood sample containing a large amount of haemocytes accurately, efficiently
Ground is sorted out.By improving isolation technics and equipment, high capture rate, high-purity and high-throughout requirement are met, so that
As the instrument that disclosure satisfy that scientific research and clinical demand.
Nano material is smaller (1-100nm) due to its size, with the properties such as special optics, electronics and magnetics, energy
Enough it is effectively applied to various medical researches.Electrostatic spinning be it is a kind of can with it is fast and convenient prepare with high-specific surface area nanometer
Or the production technology of micron order fiber, this method has the characteristics that：Preparation process is simple, kinds of fibers is various；Obtained
Nanofiber has great specific surface area, can provide substantial amounts of cells contacting site, makes the number of capture cell in unit area
Amount increase [Chen, L., et al., Aptamer-Mediated Efficient Capture and Release of T
Lymphocytes on Nanostructured Surfaces.Advanced Materials,2011,23,4376-4380]。
Numerous studies show that nanofiber easily carries out surface modification, and can load bioactive molecule, such as protein, nucleic acid, sugar
Class and growth factor etc..Meanwhile, nanofiber can simulate natural extracellular matrix (Extracellular matrix, ECM),
Good microenvironment is provided for the adhesion of cell, propagation and physiological function.
At present, the platform for capturing CTCs using the nano fibrous membrane of antibody modification is favored by researcher, utilizes function
The report that change nanofiber is captured to cell is also more and more.Wuhan University Zhang etc. [Zhang, N., et al.,
Electrospun TiO2Nanofiber-Based Cell Capture Assay for Detecting Circulating
Tumor Cells from Colorectal and Gastric Cancer Patients.Advanced Materials,
2012,24,2756-2760] using functionalized titanic oxide nanometer Electrospun coating substrate, the separation to CTCs in blood is realized
Enrichment.Except functionalization recognizes antibody, the Electrospun of substrate deposition is arranged in parallel, its topological structure and extracellular matrix phase interaction
With to improve CTC capture rates.Research shows, cell surface nanostructured component (microvillus and filopodia) and extracellular base
Matter interacts, the function such as influence cell adhesion, active transport and differentiation.Base outside nano material analog cell thus can be passed through
Texture builds the Nanosurface of some controllable cell adhesions, realizes the capture to CTC.
Because micro-fluidic chip line size is matched very much in micron dimension and cell size, stream can be manipulated well again
Body, is highly suitable to be applied for cell sorting.Moreover, sorting CTCs sample is very limited, handle micro using micro-fluidic chip
Fluid just compensate for this defect, so by the common concern of researcher.In the last few years, there is increasing researcher will
Microfluidic chip technology is applied in CTCs sorting.
Nano material is combined with micro-fluidic chip, cell can be further improved with capturing the contact of target spot, so that
Solve the problems such as enrichment sensitivity is relatively low when few pending blood sample amount in clinical detection and single use micro-fluidic chip.Zhao
Et al. [Zhao, L., et al., High-purity prostate circulating tumor cell isolation by
a polymer nanofiber-embedded microchip for whole exome sequencing.Adv Mater,
2013.25(21):P.2897-2902 the micro-fluidic chip that there is EpCAM specific antibodies to modify on surface] is built, passes through control
The factor such as flow rate of liquid in microchannel length, passage, adds contact of the cell surface identification probe with cell, further carries
High CTC acquisition sensitivity.And the substrate can realize single celled micro-dissections, and carry out full base to the CTCs of acquisition
Because of sequencing, good effect is shown.However, EpCAM antibody can only capture the cancer cell of epithelial origin, and it is expensive, no
It is easy to maintain, with very big application limitation.
Folic acid is as indispensable vitamin in a kind of human body, with very important biological significance.In recent years
Research shows that folacin receptor is in the malignant cell surface height expression of epithelium and non-epithelial origin, folic acid (Folic
Acid, FA) there is very strong specificity and affinity between part and folacin and folacin receptor (FR), far above antibody
Affinity between acceptor, thus folate conjugate detects the tumour of FR positive expressions frequently as targeting diagnosis reagent.Ge Nuo
Si Bo companies use the nucleic acid molecular probe of modified with folic acid as targeting agent, and cancer patient is detected using round pcr is targetted
CTC levels in blood, the supplementary means diagnosed as non-small cell lung cancer CT so that the accuracy rate of diagnosis is greatly improved, mesh
It is preceding to have obtained CFDA certifications, preferable effect is obtained in the early diagnosis of lung cancer.Compared with the nanofiber of antibody modification,
The nanofiber adhesive force of folic acid functionalization is strong, cheap, is easy to store and transports, so the non-antibody target of such as folic acid class
There is very important commercial value in following in-vitro diagnosis field to molecule.Micro-fluidic chip can be grasped in micron order yardstick
Flow control body, has great advantage in cell capture, and CTCs capture is carried out in a dynamic condition, process is advantageously implemented
Automation and industrialization.
Retrieve domestic and international pertinent literature and patent results show：The nanofiber capture folacin receptor height expression of modified with folic acid
Cell simultaneously combines the technology that microfluidic chip technology carries out cell capture, and there is not been reported.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of nanometer for being used to capture folacin receptor height expression cancer cell
Tunica fibrosa and its preparation method and application, the nano fibrous membrane is modified using folate-targeted, with reference to the Large ratio surface of nanofiber
The advantage of product and microfluidic chip technology to cell and fluid precise manipulation, it is ensured that high capture rate, stability of material is good, uses
It is convenient, make simple, it is with low cost, with good application prospect.
The invention provides a kind of nano fibrous membrane for being used to capture folacin receptor height expression cancer cell, the nanofiber
Film is the poly lactic-co-glycolic acid PLGA nano fibrous membranes of modified with folic acid；Wherein, the folic acid is connected to polyamino chemical combination in advance
Thing surface.
The multiamino compound is polyethyleneimine PEI.
It is crosslinked during the modification using EDC/NHS.
Fibre diameter in the nano fibrous membrane is between 10nm~2000nm.
Present invention also offers a kind of preparation method for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell,
Including：
(1) poly lactic-co-glycolic acid PLGA is dissolved in organic solvent, stirring obtains spinning solution；Then carry out Static Spinning
Silk, dries, obtains PLGA nano fibrous membranes；
(2) folic acid is dissolved in organic solvent, reacted 2-3 days with PEI after being activated through EDC/NHS, dialysis is freeze-dried
To PEI-FA (as illustrated in figure 1 c)；Wherein, folic acid, EDC, NHS and PEI mass ratio are 4-8:20-50:10-20:200-500；
(3) in the PLGA nano fibrous membranes obtained using EDC/NHS activation steps (1), immersion ultra-pure water, it is subsequently added
Tunica fibrosa is taken out after the PEI-FA aqueous solution, reaction 2-3d, cleans, dries in immersion ultra-pure water, produce for capturing folic acid
The nano fibrous membrane (as shown in Figure 1 d) of acceptor height expression cancer cell；Wherein, PLGA nano fibrous membranes, EDC, NHS and PEI-FA
Mass ratio be 100:90-96:50-60:2-10.
The mass volume ratio of PLGA and organic solvent in the step (1) are 1g:4mL；PLGA Mw is 80,000.
Organic solvent in the step (1) is volume ratio 3:The mixing of 1 tetrahydrofuran and N,N-dimethylformamide
Solution；Organic solvent in the step (2) is dimethyl sulfoxide (DMSO) DMSO.
Electrostatic spinning process parameter in the step (1) is：20-35 DEG C of temperature, voltage is 15-20kV, receives distance
For 15cm, flow velocity is 0.3mL/h, and ambient humidity is 40-50%, is received in standard glass slide or diameter 14mm circular cover glass
On.
Drying in the step (1) is natural air drying, and drying time is 12-24h.
PEI in the step (2) uses fluorescein isothiocynate FITC or other fluorochrome labels, is easy to detection.
EDC/NHS soak times in the step (2) are 2-3h.
The concentration of the aqueous solution of PEI-FA in the step (3) is 1~2mg/mL.
It is described present invention also offers a kind of application for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell
Nanofiber film combination micro-fluidic chip carries out the specificity capture of folacin receptor height expression cancer cell.
The micro-fluidic chip includes：The cell capture passage being made up of substrate of glass, cap rock, the one of cell capture passage
Hold as sample injection port, the other end is sample export, and cap rock has cylindroid array, and nano fibrous membrane is sandwiched in substrate of glass and lid
Sandwich structure is formed in the middle of layer, passes through external Boards wall and sealing passage.
The method that the nano fibrous membrane of the present invention is embedded into inside micro-fluidic chip includes：
(1) nano fibrous membrane is directly received in substrate of glass or PDMS substrate surfaces；
(2) cut when nano fibrous membrane is bonded according to long 28mm, width 8mm size with scalpel, redundance operation
Knife is gently wiped off, then clean with alcohol wipe, is dried.
(3) it is liquid leakage caused by avoiding pressure excessive, micro-fluidic core necessary is fixed as far as possible rather than with fixture.
The method of capture tumour cell includes：
(1) targeting cell is subjected to fluorescent staining, then carries out resuspension with cell culture fluid；
(2) will be first full of with phosphate buffer (PBS) in chip, then by blood sample inject micro-fluidic chip cell
Capture in passage, then rinsed with phosphate buffer (PBS)；
(3) in capture passage, because fiber surface is modified with substantial amounts of folic acid, table can be crossed with target capture folacin receptor
The cancer cell reached.When cancer cell flows through from fiber surface, targeting cell can be captured, and stick to fiber surface, non-targeted thin
Born of the same parents are with fluid outlet passage；
(4) in fluorescence microscopy Microscopic observation capture region, the targeting cell that captures can be detected and counted.
Beneficial effect
(1) present invention prepares nano fibrous membrane using electrostatic spinning technique, it is possible to achieve FA simple and efficient modification, can be with
Realize inside embedding, such as micro-fluidic chip；
(2) present invention can realize human oral cavity epithelial cancer cell using the PLGA electrostatic spinning nano fibers film of modified with folic acid
The specificity capture of (KB cell)；
(3) present invention is using folate-targeted modification, with reference to the bigger serface and microfluidic chip technology pair of nanofiber
The advantage of cell and fluid precise manipulation, it is ensured that high capture rate, stability of material is good, easy to use, makes simple, cost is low
It is honest and clean, with good application prospect.
Brief description of the drawings
Fig. 1 a-d are the synthesis schematic diagram of functionalized nano-fiber of the present invention；
The SEM figures (left side) and diameter distribution profile (right side) for the PLGA electrostatic spinning nano fibers that Fig. 2 is prepared for the present invention；
The work(of UV-visible absorption spectrum (left side) and the PEI-FI modification for the PEI-FI solution that Fig. 3 is prepared for the present invention
The shows fluorescent microscopy images (right side) of nanofiber after energyization；
The PLGA electrostatic spinning nano fibers (3) that Fig. 4 is prepared for the present invention, the PLGA nanofibers (2) after PEI is modified
With the infrared spectrogram of the PLGA nanofibers (1) after PEI, FA modification；
Fig. 5 for the present invention prepare FA modification PLGA electrostatic spinning nano fibers in different time (10min, 30min,
50min, 70min) to FA acceptors high expressing cell (KB cells) (left side) and the effect of low expression cell (L929 cells) (right side) capture
Rate；
The micro flow control chip device figure captured for cancer cell for the embedded nano fiber that Fig. 6 assembles for the present invention；
Fig. 7 is used for the situation that cancer cell is captured for the micro-fluidic chip of embedded nano fiber under the conditions of different in flow rate；
Fig. 8 is the structural representation of nano fibrous membrane of the present invention.
Embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than limitation the scope of the present invention.In addition, it is to be understood that after the content of the invention lectured has been read, people in the art
Member can make various changes or modifications to the present invention, and these equivalent form of values equally fall within the application appended claims and limited
Scope.
Embodiment 1
2.0g PLGA is weighed, THF and DMF mixed solvent (THF (tetrahydrofuran) and DMF (N-N- diformazans is dissolved in
Base formamide) volume ratio be 3:1) in, under magnetic agitation effect, until PLGA, all dissolving is finished, and obtains spinning solution dense
Spend for 25% (1g/4mL).Then spinning solution is drawn in syringe, electrostatic spinning is carried out using high-voltage electrostatic spinning machine, spun
Strand part is：25 DEG C of temperature, humidity 40-50%, voltage 20Kv；Receive apart from 15cm；Fluid flow 0.3mL/h.Wherein receive
Plate is clean slide or the circular cover glass sticked on slide.It is put into fume hood and dries 12-24h.Scanning electricity
Sub- microscope (Scanning Eelectron Microscope, SEM) result shows that the nanofiber surface of acquisition is smooth, directly
Footpath is uniform, and average fibre diameter is 850.2nm (Fig. 2).
Then PEI surfaces are arrived in FA modifications, 4.41mg FA is dissolved in 5mL dimethyl sulfoxide (DMSO) DMSO, using super
Sound about 30min is completely dissolved it, is then added dropwise under magnetic stirring after 1mL EDC aqueous solution 28.7mg, reaction 30min
1mL NHS aqueous solution 16.4mg are added, 2.5h, fully activation folacin-carboxyl group are stirred using magnetic agitation instrument.Then plus
Enter 5mL PEI dimethyl sulphoxide solution (250mg), react 2~4 days, finally dialyse under the conditions of magnetic agitation, remove free
FA, freeze-dried back.
First slide or cover glass that spinning there are PLGA nanofibers (100mg) are placed on staining rack, then leaching completely
Not in the staining jar equipped with ultra-pure water, 5mL EDC (95.8mg) aqueous solution is added, 30min is reacted under the conditions of magnetic agitation,
Then 5mL NHS (57.9mg) aqueous solution is added, at normal temperatures with activation nano fibrous membrane carboxyl 2.5h under the conditions of magnetic agitation,
The 10mL PEI-FA 13mg aqueous solution is eventually adding in staining jar, 2~3d is stirred using magnetic agitation instrument.After the completion of reaction,
Dyeing is placed in the solution of ultra-pure water, cleaning in three hours is soaked under the conditions of magnetic agitation and removes free PEI, per hour
Change water once.Wash away be positioned over after free PEI-FA, vacuum drying it is standby at low temperature.Fig. 3 ultraviolet spectrogram and fluorescence shows
Micro mirror figure, which demonstrates PEI, can successfully modify nanofiber surface, and Fig. 4 infrared spectrogram further demonstrates that polyethyleneimine
Amine-folic acid (PEI-FA) can modify nanofiber surface.
Embodiment 2
In order to verify that fibrous material has specific capture effect, with the human oral cavity epithelial with folacin receptor height expression
Cell (KB cells) is model cell to examine the functionalized nano-fiber specificity that the FA of preparation is modified to capture the effect of KB cells
Really.
The circular cover glass for obtaining the covering nanofiber that FA has been modified is removed from slide, sample is then put into 12
Well culture plate.The sample of different time points is placed in different culture plates, each 3 Duplicate Samples of sample.After ultraviolet irradiation 1h,
400 μ L serum free mediums immersion fiber slide 30min is added in the orifice plate.Then culture medium is suctioned out, by ready without blood
Clear cell suspension is added in orifice plate, and cell concentration is 250/mL, and suspension dosage is 400 μ L, it is ensured that added in each orifice plate
Cell be 100.Then stand 10,20,40,60min, then take out culture medium, and with PBS three times, using it is living carefully
Born of the same parents' dyestuff calcein is dyed, and is counted in fluorescence microscopy Microscopic observation.Then cell capture efficiency is counted.Control group uses L929
Cell (cell of folacin receptor low expression), is handled using above-mentioned similarity condition.The Nanowire of functional modification is understood in figure
The literalness PLGA nanofibers of peacekeeping have significant difference to the capture rate of KB cells, and compare work(knowable to the figure of left and right
Nanofiber can be changed to the capture rate of folacin receptor high expressing cell apparently higher than low expression cell (Fig. 5).
Embodiment 3
Prepare dimethyl silicone polymer (PDMS) chip (as shown in Figure 6)：By performed polymer and curing agent with mass ratio 10：1
Ratio is well mixed, vacuum degassing bubble, and PDMS solution is poured into and is placed with the culture dish of silicon chip mould, solid in 60 DEG C of baking ovens
Change.The PLGA nano fibrous membranes for modifying folic acid are cut into the size close with chip cell capture passage, then will be cut
The slide of nano fibrous membrane together carries out ion processing with micro-fluidic chip.Wherein, it is covered with the slide of nano fibrous membrane only
Glass surface is handled, tunica fibrosa is blocked temporarily using slightly larger PDMS films, prevents destruction of the plasma to tunica fibrosa.Processing
The two is bonded under a certain pressure rapidly afterwards.Device after bonding is wrapped into diaphragm seal prevents pollution, is kept in dark place.Cell is caught
Obtain before experiment and to be stained with medical infusion lines in feeder connection and outlet using epoxide resin AB glue, and clamp with fixture packaged core
Piece, reduces risk of leakage.
Micro-fluidic chip captures circulating tumor cell：
Step A：Suspended after KB cell pancreatin is digested, 1000rpm is centrifuged off pancreatin, then outstanding with serum free medium
It is floating, lucifuge dyeing 30min under the conditions of calcein live cell fluorescent dye, 37 DEG C is then added in culture medium, is centrifuged off
Dyeing liquor, then, finally recycles serum free medium suspension cell with PBS twice, taken respectively after counting containing 125,250,
The μ L of suspension 10 of 500 KB cells；
Step B：It will be first full of with phosphate buffer (PBS) in chip, then the mixed cell suspension 1mL prepared noted
Enter in the fiber target capture passage of micro-fluidic chip, flow velocity is about 1-5mL/h, phosphate buffer (PBS) is then used again
Rinse；
Step C：, can be with target capture folacin receptor because fiber surface is modified with substantial amounts of folic acid in capture passage
The cancer cell of overexpression, when cell flows through from fiber surface, cell can be caught in tunica fibrosa surface flow, targeting cell
Obtain, stick to fiber surface, non-targeted cell is flowed into liquid storage tank with fluid outlet passage, bottom is deposited on by Action of Gravity Field
Portion；
Step D：Capture region is detected under fluorescence microscope, the targeting cell captured can be detected.Sent out after statistics
The chip obtains the maximum capture rate of KB cells up to more than 92% (Fig. 7) when existing flow velocity is 1mL/h.
Claims (10)
1. a kind of nano fibrous membrane for being used to capture folacin receptor height expression cancer cell, it is characterised in that：The nano fibrous membrane
For the poly lactic-co-glycolic acid PLGA nano fibrous membranes of modified with folic acid；Wherein, the folic acid is connected to multiamino compound in advance
Surface.
2. a kind of nano fibrous membrane for being used to capture folacin receptor height expression cancer cell according to claim 1, its feature
It is：The multiamino compound is polyethyleneimine PEI.
3. a kind of nano fibrous membrane for being used to capture folacin receptor height expression cancer cell according to claim 1, its feature
It is：It is crosslinked during the modification using EDC/NHS.
4. a kind of preparation method for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell, including：
(1) poly lactic-co-glycolic acid PLGA is dissolved in organic solvent, stirring obtains spinning solution；Electrostatic spinning is then carried out, is done
It is dry, obtain PLGA nano fibrous membranes；
(2) folic acid is dissolved in organic solvent, reacted 2-3 days with PEI after being activated through EDC/NHS, dialysis, freeze-drying is obtained
PEI-FA；Wherein, folic acid, EDC, NHS and PEI mass ratio are 4-8:20-50:10-20:200-500；
(3) in the PLGA nano fibrous membranes obtained using EDC/NHS activation steps (1), immersion ultra-pure water, it is subsequently added PEI-FA
The aqueous solution, tunica fibrosa is taken out after reaction 2-3d, cleans, dry in immersion ultra-pure water, produce high for capturing folacin receptor
Express the nano fibrous membrane of cancer cell；Wherein, PLGA nano fibrous membranes, EDC, NHS and PEI-FA mass ratio are 100:90-
96:50-60:2-10。
5. a kind of preparation side for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell according to claim 4
Method, it is characterised in that：Organic solvent in the step (1) is volume ratio 3:1 tetrahydrofuran and N,N-dimethylformamide
Mixed solution；Organic solvent in the step (2) is dimethyl sulfoxide (DMSO) DMSO.
6. a kind of preparation side for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell according to claim 4
Method, it is characterised in that：Electrostatic spinning process parameter in the step (1) is：20-35 DEG C of temperature, voltage is 15-20kV, is connect
It is 15cm to receive distance, and flow velocity is 0.3mL/h, and ambient humidity is 40-50%.
7. a kind of preparation side for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell according to claim 4
Method, it is characterised in that：PEI in the step (2) is marked using fluorescein isothiocynate FITC.
8. a kind of preparation side for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell according to claim 4
Method, it is characterised in that：The concentration of the aqueous solution of PEI-FA in the step (3) is 1~2mg/mL.
9. a kind of application as claimed in claim 1 for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell, its
It is characterised by：The nanofiber film combination micro-fluidic chip carries out the specificity capture of folacin receptor height expression cancer cell.
10. a kind of application for being used to capture the nano fibrous membrane of folacin receptor height expression cancer cell according to claim 9,
It is characterized in that：The micro-fluidic chip includes：The cell capture passage being made up of substrate of glass, cap rock, cell capture passage
One end be sample injection port, the other end is sample export, and cap rock has cylindroid array, and nano fibrous membrane is sandwiched in substrate of glass
With formation sandwich structure in the middle of cap rock, pass through external Boards wall and sealing passage.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103040724A (en) * | 2012-12-22 | 2013-04-17 | 中国科学院深圳先进技术研究院 | Nano drug delivery system containing polymer and phospholipid and preparation method thereof |
| CN104790216A (en) * | 2015-04-17 | 2015-07-22 | 东华大学 | Method for preparing folic acid functional nanofibers for target capture of cancer cells |
| CN105854966A (en) * | 2016-06-14 | 2016-08-17 | 东华大学 | Microfluidic chip embedding functional nanofiber membrane and application of microfluidic chip |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103040724A (en) * | 2012-12-22 | 2013-04-17 | 中国科学院深圳先进技术研究院 | Nano drug delivery system containing polymer and phospholipid and preparation method thereof |
| CN104790216A (en) * | 2015-04-17 | 2015-07-22 | 东华大学 | Method for preparing folic acid functional nanofibers for target capture of cancer cells |
| CN105854966A (en) * | 2016-06-14 | 2016-08-17 | 东华大学 | Microfluidic chip embedding functional nanofiber membrane and application of microfluidic chip |
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