CN107653179A - A kind of micropore duct device and apply its circulating tumor cell method for catching - Google Patents
A kind of micropore duct device and apply its circulating tumor cell method for catching Download PDFInfo
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- CN107653179A CN107653179A CN201610596857.3A CN201610596857A CN107653179A CN 107653179 A CN107653179 A CN 107653179A CN 201610596857 A CN201610596857 A CN 201610596857A CN 107653179 A CN107653179 A CN 107653179A
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Abstract
The present invention relates to the technical field of medical treatment detection, discloses a kind of micropore duct device and applies its circulating tumor cell method for catching.The micropore duct device includes：Substrate, the microchannel formed on substrate；The pillar array being arranged on microchannel bottom surface, and it is respectively perpendicular the nanometer stick array being grown on microchannel bottom surface, side wall and/or each cylinder；Also include the encapsulated layer of encapsulation microchannel, and inlet and liquid outlet are provided with encapsulated layer.In the above-mentioned technical solutions, by setting the nanometer stick array of geometry lattice arrangement inside micropore duct device, whole blood sample flows through between nanometer stick array, adds circulating tumor cell with catching the frequency of interracial contact, can increase capturing efficiency.In addition, microchannel installation cost is low, it is easy to mass production.Also, simple to operate using device seizure circulating tumor cell, sample usage amount is few, takes short.
Description
Technical field
The present invention relates to medical treatment detection technical field, more particularly to a kind of micropore duct device and apply its circulation
Tumour cell method for catching.
Background technology
Cancer metastasis is the most fearful one side of cancer, and the cancer mortality more than 90% is as caused by transfer.Cancer turns
Shifting refers to that tumour cell sheds into blood from home position and forms circulating tumor cell, and circulating tumor cell falls with blood flow
To new position and form metastatic tumor.Therefore circulating tumor cell carries the information of this primary tumor, how effectively to catch blood
In circulating tumor cell early-stage cancer detect and cancer metastasis research in terms of it is significant.
But the content due to circulating tumor cell in blood is extremely low, seizure rapidly and efficiently, separation are realized to it still
It is so a great problem.CellSearch of the conventional method such as based on immuno magnetic cell separation technology is unique U.S. FDA approval
For the clinical technology for catching circulating tumor cell, it by nano magnetic particle pan coating antibody (Anti-EPCAM) with
Circulating tumor cell is specifically bound, and circulating tumor cell is isolated under magnetic field or so, but separative efficiency is relatively low.
The content of the invention
The invention provides a kind of micropore duct device and its application method, improves the effect for catching circulating tumor cell.
The invention provides a kind of micropore duct device, the micropore duct device includes：Substrate, form on the substrate micro-
Duct；The pillar array being arranged on the microchannel bottom surface, and vertical-growth the microchannel bottom surface, side wall and/or
Nanometer stick array on each cylinder；Also include the encapsulated layer for encapsulating the microchannel, and feed liquor is provided with the encapsulated layer
Mouth and liquid outlet.
In the above-mentioned technical solutions, by setting the nanometer stick array of geometry lattice arrangement inside micropore duct device, entirely
Blood sample flows through between nanometer stick array, adds circulating tumor cell and catches the frequency of interracial contact, can increase seizure effect
Rate.The seizure interface is nanorod surfaces, substantially increases the surface area for catching interface, enhances and catches interface and circulating tumor
The interaction of cell, further improves capturing efficiency.In addition, microchannel installation cost is low, it is easy to mass production.Also,
Simple to operate using device seizure circulating tumor cell, sample usage amount is few, takes short.
In specific set, for the height of the cylinder between 10-200 μm, diameter is adjacent between 10-100 μm
Cylinder spacing between 20-200 μm.
In specific set, the cylinder is shaped as cylinder, quadrangular, six prisms or eight prisms.
In specific set, the nanorod length between 200-2000nm, diameter between 20-200nm it
Between.
In specific set, for the width of the microchannel between 0.5-10mm, length is deep between 10-200mm
Degree is between 10-200mm.
Nanometer rods therein are titanium dioxide nano-rod, zinc oxide nano rod or silicon nanorod.
Substrate therein is silicon substrate, glass substrate or dimethyl silicone polymer substrate.
Circulating tumor cell specific antibody is attached with nanometer rods wherein in the microchannel.
Present invention also offers a kind of application method of micropore duct device, this method comprises the following steps：
Circulating tumor cell specific antibody in attachment in nanometer rods in microchannel；
Whole blood sample containing circulating tumor cell is flowed through into micropore duct device, carries out circulating tumor cell seizure；
The non-specific cell caught is cleaned after seizure with phosphate buffer.
In the above-mentioned methods, by setting the nanometer stick array of geometry lattice arrangement, whole blood inside micropore duct device
Product flow through between nanometer stick array, add circulating tumor cell with catching the frequency of interracial contact, can increase capturing efficiency.Should
Seizure interface is nanorod surfaces, substantially increases the surface area for catching interface, enhances and catches interface and circulating tumor cell
Interaction, further improve capturing efficiency.In addition, microchannel installation cost is low, it is easy to mass production.Also, use
Device seizure circulating tumor cell is simple to operate, and sample usage amount is few, takes short.
In specific steps, the whole blood sample containing circulating tumor cell is flowed through described with 0.1-10mL/h speed
Microchannel.
In concrete operations, circulating tumor cell specific antibody is specific in attachment in the above-mentioned nanometer rods in microchannel
For：
By Miniature injection pump successively by 4% γ-mercaptopropyl trimethoxysilane ethanol solution, 25mM4- maleimides
Base butyric acid-N- succinimide esters, 10 μ g/mL Streptavidin, the circulating tumor cell of 10 μ g/ml biotin labelings are special
Property antibody flow through micropore duct device, make to adhere to upper specific antibody in nanometer rods.
This method also includes carrying out increment culture after the circulating tumor cell of seizure is taken off in micropore duct device,
For the further analysis detection to circulating tumor cell.
Brief description of the drawings
Fig. 1 is the microchannel schematic device that example of the present invention provides；
Fig. 2 is the pillar array and cylinder surface sweeping electron microscopic picture inside the microchannel that example of the present invention provides；
Fig. 3 is the titanium dioxide nano-rod of the pillar array and column vertical growth inside the microchannel that example of the present invention provides
Surface sweeping electron microscopic picture；
Fig. 4 is the circulating tumor cell immunofluorescence dye in the whole blood sample that the micropore duct device that example of the present invention provides is caught
Chromatic graph；
Fig. 5 is the circulating tumor cell ESEM in the whole blood sample that the micropore duct device that example of the present invention provides is caught
Figure.
Reference：
1- substrate 2- microchannel 21- cylinder 22- nanometer rods
3- encapsulated layer 31- inlet 32- liquid outlets
Embodiment
In order that the object, technical solutions and advantages of the present invention are clearer, the present invention is made below in conjunction with accompanying drawing into
One step it is described in detail, it is clear that described embodiment is only part of the embodiment of the present invention, rather than whole implementation
Example.Based on the embodiment in the present invention, what those of ordinary skill in the art were obtained under the premise of creative work is not made
All other embodiment, belongs to the scope of protection of the invention.
As shown in Figure 1, Figure 2 and Figure 3, Fig. 1 shows that, the embodiments of the invention provide a kind of micropore duct device, Fig. 2 is this
The pillar array and cylinder surface sweeping electron microscopic picture inside microchannel that example provides；Fig. 3 is the microchannel that example of the present invention provides
Internal pillar array and the titanium dioxide nano-rod surface sweeping electron microscopic picture of column vertical growth；
The embodiments of the invention provide a kind of micropore duct device, the micropore duct device includes：Substrate 1, formed in the base
Microchannel 2 on plate 1；The pillar array being arranged on the bottom surface of microchannel 2, and vertical-growth is at the bottom of microchannel 2
Nanometer stick array on face, side wall and/or each cylinder 21；Also include the encapsulated layer 3 for encapsulating the microchannel 2, and the envelope
Inlet 31 and liquid outlet 32 are provided with dress layer 3.
Setting nanometer that in the above-described embodiments can only on the bottom surface of microchannel 2 either side wall or each cylinder 21
Rod array, blood is set partly to be contacted when flowing through microchannel 2 with nanometer rods 22.Can also be in the bottom surface of microchannel 2, side wall and each
Nanometer stick array is respectively provided with cylinder 21, blood is all contacted when flowing through microchannel 2 with nanometer rods 22.
In the above-described embodiments, the seizure of circulating tumor cell is carried out by providing a kind of micropore duct device, and had
When body is set, by setting pillar array in microchannel 2, and growth forms nanometer in pillar array, the side wall of microchannel 2
Rod array, so as to increase the contact area of microchannel 2 and whole blood sample, that is, seizure interface and the circulation for increasing micropore duct device are swollen
The contact frequency of oncocyte, the seizure interface are the surface of nanometer rods 22.Pass through the cylinder 21 of setting, nanometer in microchannel 2
Rod 22 adds the frequency of contact, enhances the interaction for catching interface and circulating tumor cell, further improves and catches effect
Rate.
Microchannel 2 therein is the microchannel 2 for etching formation on substrate 1, i.e., is performed etching on the material of substrate 1, shape
Into pore passage structure, specifically, the substrate 1 in selection, can use different materials, such as：Silicon substrate, glass substrate or poly- two
The common substrate such as methylsiloxane substrate.In specific make, two inches of substrate 1 in acetone, isopropanol and is gone successively
It is cleaned by ultrasonic in ionized water, is then dried up with nitrogen.By sun-9i photoresists paving on a silicon substrate, then obtained using photoetching technique
To the structure of microchannel 2.In the microchannel structure of preparation, the data of microchannel 2 meet：Width is long between 0.5-10mm
Degree is between 10-200mm, and depth is between 10-200mm.And the length and width of the microchannel 2 deeply can be according to the experiment of reality
Situation carries out different settings, but needs the requirement for meeting above-mentioned data interval.
Cylinder 21 therein is the column structure that formation is etched on microchannel 2, specifically, the etching shape in microchannel 2
Into the structure of cylinder 21, pillar array is obtained, and the pillar array formed meets：Height between 10-200 μm, diameter between
Between 10-100 μm, the spacing of adjacent cylinder 21 is between 20-200 μm.The size of specific cylinder 21 can be according to reality
The experimental conditions on border carry out different settings, but need the requirement for meeting above-mentioned data interval.In addition, the shape of the cylinder 21 can
Think the different shapes of cylinder 21, such as cylinder, quadrangular, six prisms, the cylinder 21 of eight prism arbitrary shapes.
In specific set, the nanometer rods 22 are using the nanometer rods being made with biocompatibility nano material, tool
Body, the materials of the nanometer rods 22 is zinc oxide, silicon, titanium dioxide etc., i.e., the nanometer rods 22 are that titanium dioxide nano-rod, silicon are received
Rice rod or zinc oxide nano rod etc., above-mentioned nanometer rods 22 have a high-ratio surface, high cellular affinity, and close with biomolecule
Size the features such as, so as to preferably capture circulating tumor cell, improve the seizure effect of circulating tumor cell.And should
Nanometer rods 22 are the growth formation on the bottom surface of microchannel 2, side wall and cylinder 21, by taking titanium dioxide as an example, by titanium dioxide kind
Sublayer is by sputtering vacuum deposition in the side wall of microchannel 2, bottom surface and silicon column sidewall surfaces, and annealing is 1 small under the conditions of 500 DEG C
When, then substrate is put into butyl titanate, deionized water and concentrated hydrochloric acid (36.5%-38% weight) mixed liquor.And poly- four
Reacted in PVF hydrothermal reaction kettle, vertical-growth goes out nanometer rods on the surface of the side wall of microchannel 2, bottom surface and cylinder 21
Array.And when being formed, the length of nanometer rods 22 is between 200-2000nm, and diameter is between 20-200nm.
By foregoing description as can be seen that the micropore duct device by particular design that the present embodiment provides, inside is several
The post array of what lattice arrangement, whole blood sample flow through between pillar array, add circulating tumor cell with catching interracial contact
Frequency, capturing efficiency can be increased.Hung down importantly, being one layer in the side wall of microchannel 2, bottom surface and silicon column sidewall surfaces
The nanometer stick array of growing straight length, substantially increases the specific surface area for catching interface, enhances and catches interface and circulating tumor cell
Interaction, further improve capturing efficiency.
In addition, microchannel installation cost is low, it is easy to mass production.The silicon of product used in micropore duct device of the present invention
The prices such as piece, glass, dimethyl silicone polymer are relatively low.Technique used is photoetching, and plasma etching, hydro-thermal reaction etc. is standard
Change flow to be easy to produce in batches.
And the micropore duct device that the present embodiment provides has preferable bio-compatible using nanometer stick array as interface is caught
Property, do not influence the further culture and analysis detection of circulating tumor cell captured.
The embodiment of the present invention additionally provides a kind of application method of micropore duct device, and carrying out circulating tumor cell using it catches
Method is caught, this method comprises the following steps：
Circulating tumor cell specific antibody in attachment on the surface of nanometer rods 22 in microchannel 2；
The whole blood sample containing circulating tumor cell is flowed through into microchannel with certain speed by microsyringe etc. to fill
Put, carry out circulating tumor cell seizure；
The non-specific cell caught is cleaned after seizure with phosphate buffer.
In order to facilitate the above method for understanding that the present embodiment provides, it is entered with reference to specific accompanying drawing and embodiment
Row detailed description.
This method includes：
Step 1：Circulating tumor cell specific antibody in attachment on the surface of nanometer rods 22 in microchannel 2；
Specifically, by Miniature injection pump successively by 4% γ-mercaptopropyl trimethoxysilane ethanol solution, 25mM4- horses
Come imide butyric acid-N- succinimide esters, 10 μ g/mL Streptavidin, the circulating tumor of 10 μ g/ml biotin labelings
Cell-specific antibodies flow through micropore duct device, make to adhere to upper specific antibody in nanometer rods 22.
Step 2：Whole blood sample containing circulating tumor cell is flowed through into microchannel, carries out circulating tumor cell seizure；
Can by microsyringe etc. by the whole blood sample containing circulating tumor cell with 0.1-10mL/h speed stream
Cross microchannel.
Step 3：Increment culture is carried out after the circulating tumor cell of seizure is taken off in micropore duct device, is used for
Further analysis detection to circulating tumor cell.
In the above-mentioned methods, by setting the nanometer stick array of geometry lattice arrangement, whole blood inside micropore duct device
Product flow through between nanometer stick array, add circulating tumor cell with catching the frequency of interracial contact, can increase capturing efficiency.Should
Seizure interface is the surface of nanometer rods 22, substantially increases the surface area for catching interface, enhances seizure interface and circulating tumor is thin
The interaction of born of the same parents, further improves capturing efficiency.In addition, microchannel installation cost is low, it is easy to mass production.Also, make
Simple to operate with device seizure circulating tumor cell, sample usage amount is few, takes short.
In order to facilitate the micro-channel device and its application method for understanding that the present embodiment provides, with reference to specific example pair
It is described in detail.
Embodiment 1：
1. liang inch silicon wafer is cleaned by ultrasonic 10 minutes in acetone, isopropanol and deionized water successively, then blown with nitrogen
It is dry.By sun-9i photoresists paving on a silicon substrate, then the structure of microchannel 2 is obtained using photoetching technique.Use plasma etching
20min obtains wide 2mm, long 100mm, deep 100um microchannel 2, the height that hexagonal lattice arranges is into inside microchannel 2Diameter, spacingSilicon column array, as shown in Figure 2.
2. by titanium oxide seed layer by sputtering vacuum deposition in the side wall of microchannel 2, bottom surface and silicon column sidewall surfaces,
And annealed 1 hour under the conditions of 500 DEG C, then substrate is put intoButyl titanate, 6 ml deionized waters, 14 milliliters of dense salt
In sour (36.5%-38% weight) mixed liquor.And reacted 8 hours under the conditions of 140 DEG C in polytetrafluoroethylene (PTFE) hydrothermal reaction kettle,
Obtain the titanic oxide nanorod array of the vertical interface growth of microchannel 2, diameter 20nm, length 2000nm, as shown in Figure 3.
3. substrate is encapsulated with dimethyl silicone polymer (PDMS), and inlet 31 and liquid outlet 32 are stayed at both ends.
4. with Miniature injection pump successively by 4% γ-mercaptopropyl trimethoxysilane ethanol solution, 25mM4- maleimides
Base butyric acid-N- succinimide esters, 10 μ g/mL Streptavidin, the circulating tumor cell of 10 μ g/ml biotin labelings are special
Property antibody (anti-EpCAM) flow through micropore duct device, make the upper specific antibody of the interface of microchannel 2 attachment.
5. and then under 37 DEG C and 5% carbon dioxide conditions with Miniature injection pump by the whole blood containing circulating tumor cell
Product flow through micropore duct device 1h with 2mL/h flow velocity.Fixed with after PBS wash mills with 4% paraformaldehyde, 0.1%
Triton X-100 punch, 1% bovine serum albumin(BSA) closing, Alexa Fluor 647 mark anti-cytokeratin and
The cell of seizure is identified in the anti-CD dyeing of FITC marks under inverted fluorescence microscope, as shown in Figure 4.To the cell of seizure
Fixed with 2.5% glutaraldehyde and surface sweeping electron microscopic observation is carried out after gradient alcohol dehydration, as shown in Figure 5.
6. it is 45% that micropore duct device circulating tumor cell capturing efficiency is obtained under the experiment condition.
Embodiment 2：
1. liang inch silicon wafer is cleaned by ultrasonic 10 minutes in acetone, isopropanol and deionized water successively, then blown with nitrogen
It is dry.By sun-9i photoresists paving on a silicon substrate, then the structure of microchannel 2 is obtained using photoetching technique.Use plasma etching
35min obtains wide 10mm, long 10mm, deepMicrochannel 2, be into inside microchannel 2 hexagonal lattice arrangement height
DegreeDiameterSpacingSilicon column array, it is similar with example 1.
2. by titanium oxide seed layer by sputtering vacuum deposition in the side wall of microchannel 2, bottom surface and silicon column sidewall surfaces,
And annealed 1 hour under the conditions of 500 DEG C, then substrate is put intoButyl titanate, 16 ml deionized waters, 4 milliliters dense
In hydrochloric acid (36.5%-38% weight) mixed liquor.And reaction 2 is small under the conditions of 180 DEG C in polytetrafluoroethylene (PTFE) hydrothermal reaction kettle
When, obtain the titanic oxide nanorod array of the vertical interface growth of microchannel 2, diameter 200nm, length 200nm and the class of example 1
Seemingly.
3. substrate is encapsulated with dimethyl silicone polymer (PDMS), and inlet 31 and liquid outlet 32 are stayed at both ends
4. with Miniature injection pump successively by 4% γ-mercaptopropyl trimethoxysilane ethanol solution, 25mM 4- maleimides
Amido butyric acid-N- succinimide esters, 10 μ g/mL Streptavidin, the circulating tumor cell of 10 μ g/ml biotin labelings are special
Heterogenetic antibody (anti-EpCAM) is to flow through micropore duct device, makes the upper specific antibody of the interface of microchannel 2 attachment.
5. and then under 37 DEG C and 5% carbon dioxide conditions with Miniature injection pump by the whole blood containing circulating tumor cell
Product flow through micropore duct device 3h with 0.1mL/h flow velocity.Fixed with after PBS wash mills with 4% paraformaldehyde, 0.1%
Triton X-100 are punched, the closing of 1% bovine serum albumin(BSA), Alexa Fluor647 marks anti-cytokeratin and FITC
The cell of seizure is identified in the anti-CD dyeing of mark under inverted fluorescence microscope.The cell of seizure is consolidated with 2.5% glutaraldehyde
Surface sweeping electron microscopic observation is carried out after fixed and gradient alcohol dehydration.
6. it is 89% that micropore duct device circulating tumor cell capturing efficiency is obtained under the experiment condition.
Embodiment 3：
1. liang inch silicon wafer is cleaned by ultrasonic 10 minutes in acetone, isopropanol and deionized water successively, then blown with nitrogen
It is dry.By sun-9i photoresists paving on a silicon substrate, then the structure of microchannel 2 is obtained using photoetching technique.Use plasma etching
10min obtains wide 0.5mm, long 200mm, deep 10um microchannel 2, the height that hexagonal lattice arranges is into inside microchannel 2
10 μm, 10 μm of diameter, the silicon column array of 20 μm of spacing is similar with example 1.
2. by titanium oxide seed layer by sputtering vacuum deposition in the side wall of microchannel 2, bottom surface and silicon column sidewall surfaces
And annealed 1 hour under the conditions of 500 DEG C, then substrate is put intoButyl titanate, 10 ml deionized waters, 10 milliliters dense
In hydrochloric acid (36.5%-38% weight) mixed liquor.And reaction 6 is small under the conditions of 160 DEG C in polytetrafluoroethylene (PTFE) hydrothermal reaction kettle
When, obtain the titanic oxide nanorod array of the vertical interface growth of microchannel 2, diameter 100nm, length 1000nm, in the class of example 1
Seemingly.
3. substrate is encapsulated with dimethyl silicone polymer (PDMS), and inlet 31 and liquid outlet 32 are stayed at both ends.
4. with Miniature injection pump successively by 4% γ-mercaptopropyl trimethoxysilane ethanol solution, 25mM 4- maleimides
Amido butyric acid-N- succinimide esters, 10 μ g/mL Streptavidin, the circulating tumor cell of 10 μ g/ml biotin labelings are special
Heterogenetic antibody (anti-EpCAM) is to flow through micropore duct device, makes the upper specific antibody of the interface of microchannel 2 attachment.
5. and then under 37 DEG C and 5% carbon dioxide conditions with Miniature injection pump by the whole blood containing circulating tumor cell
Product flow through micropore duct device 1h with 1mL/h flow velocity.Fixed with after PBS wash mills with 4% paraformaldehyde, 0.1%
Triton X-100 punch, 1% bovine serum albumin(BSA) closing, Alexa Fluor 647 mark anti-cytokeratin and
The cell of seizure is identified in the anti-CD dyeing of FITC marks under inverted fluorescence microscope.To the cell of seizure with 2.5% penta 2
Aldehyde is fixed and surface sweeping electron microscopic observation is carried out after gradient alcohol dehydration.
6. it is 76% that micropore duct device circulating tumor cell capturing efficiency is obtained under the experiment condition.
By above-mentioned specific embodiment 1, embodiment 2 and embodiment 3 as can be seen that the micropore provided using the present embodiment
Duct device, by setting the nanometer stick array of geometry lattice arrangement inside micropore duct device, whole blood sample is in nanometer stick array
Between flow through, add circulating tumor cell with catch interracial contact frequency, capturing efficiency can be increased.The seizure interface is nanometer
Rod surface, the surface area for catching interface is substantially increased, enhance the interaction for catching interface and circulating tumor cell, enter one
Step improves capturing efficiency.In addition, microchannel installation cost is low, it is easy to mass production.Also, it is swollen to catch circulation using the device
Oncocyte is simple to operate, and sample usage amount is few, takes short.
Obviously, those skilled in the art can carry out the essence of various changes and modification without departing from the present invention to the present invention
God and scope.So, if these modifications and variations of the present invention are related to the model of the claims in the present invention and its equivalent technologies
Within enclosing, then the present invention is also intended to comprising including these changes and modification.
Claims (12)
- A kind of 1. micropore duct device, it is characterised in that including：Substrate, form microchannel on the substrate；It is arranged on described Pillar array on microchannel bottom surface, and nanometer of the vertical-growth on the microchannel bottom surface, side wall and/or each cylinder Rod array；Also include the encapsulated layer for encapsulating the microchannel, and inlet and liquid outlet are provided with the encapsulated layer.
- 2. micropore duct device as claimed in claim 1, it is characterised in that the height of the cylinder between 10-200 μm, Diameter is between 10-100 μm, and the spacing of adjacent cylinder is between 20-200 μm.
- 3. micropore duct device as claimed in claim 2, it is characterised in that the cylinder is shaped as cylinder, quadrangular, six ribs Post or eight prisms.
- 4. such as the micropore duct device described in claim any one of 1-3, it is characterised in that the nanorod length is between Between 200-2000nm, diameter is between 20-200nm.
- 5. such as the micropore duct device described in claim any one of 1-4, it is characterised in that the width of the microchannel between Between 0.5-10mm, length is between 10-200mm, and depth is between 10-200mm.
- 6. such as the micropore duct device described in claim any one of 1-5, it is characterised in that the nanometer rods are received for titanium dioxide Rice rod, zinc oxide nano rod or silicon nanorod.
- 7. such as the micropore duct device described in claim any one of 1-6, it is characterised in that the substrate is silicon substrate, glass Substrate or dimethyl silicone polymer substrate.
- 8. such as the micropore duct device described in claim any one of 1-7, it is characterised in that the nanometer rods in the microchannel On be attached with circulating tumor cell specific antibody.
- 9. a kind of circulating tumor cell method for catching, it is characterised in that using the microchannel described in claim any one of 1-7 Device, comprise the following steps：Circulating tumor cell specific antibody in attachment in nanometer rods in microchannel；Whole blood sample containing circulating tumor cell is flowed through into the microchannel, carries out circulating tumor cell seizure.
- 10. method as claimed in claim 9, it is characterised in that the whole blood sample containing circulating tumor cell is with 0.1- 10mL/h speed flows through the microchannel.
- 11. method as claimed in claim 9, it is characterised in that circulation is swollen in attachment in the nanometer rods in microchannel Oncocyte specific antibody is specially：It is successively that 4% γ-mercaptopropyl trimethoxysilane ethanol solution, 25mM 4- maleimidobutyric acid-N- succinyls is sub- Amine ester, 10 μ g/mL Streptavidin, the circulating tumor cell specific antibody of 10 μ g/ml biotin labelings flow through microchannel Device, make to adhere to upper specific antibody on the seizure interface of microchannel.
- 12. such as the method described in claim any one of 9-11, it is characterised in that also include the circulating tumor cell of seizure Increment culture is carried out after being taken off in micropore duct device, for the further analysis detection to circulating tumor cell.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111019900A (en) * | 2019-12-10 | 2020-04-17 | 东华大学 | Method for constructing biological functionalized fluorinated zinc hydroxide/zinc oxide composite nano forest array in micro-channel |
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| CN111019900A (en) * | 2019-12-10 | 2020-04-17 | 东华大学 | Method for constructing biological functionalized fluorinated zinc hydroxide/zinc oxide composite nano forest array in micro-channel |
| CN111019900B (en) * | 2019-12-10 | 2023-12-19 | 东华大学 | Method for constructing biological functionalized fluorozinc hydroxide/zinc oxide composite nano-forest array in micro-channel |
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