CN207596861U - Micro-fluidic tumour cell detection chip based on immunomagnetic isolation - Google Patents
Micro-fluidic tumour cell detection chip based on immunomagnetic isolation Download PDFInfo
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- CN207596861U CN207596861U CN201721465011.2U CN201721465011U CN207596861U CN 207596861 U CN207596861 U CN 207596861U CN 201721465011 U CN201721465011 U CN 201721465011U CN 207596861 U CN207596861 U CN 207596861U
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Abstract
Micro-fluidic tumour cell detection chip based on immunomagnetic isolation, including chip body, chip body includes substrate and optical cement layer forms, the optical cement layer is equipped with sample introduction tank, reagent trough a, reagent trough b and waste liquid tank, the side of optical cement layer is equipped with electrode terminal a and electrode terminal b, sample introduction tank is connected with reagent trough a by microchannel, microchannel is followed by mixing microchannel, mixing microchannel is followed by separation microchannel, separation microchannel is followed by dyeing counting microchannel, the separation microchannel is detour type microchannel, separation is embedded with magnetic coil below microchannel, mix microchannel, it detaches and micro-pillar array is equipped in the channel of microchannel and dyeing counting microchannel, micro-pillar array is arranged by several miniature cylinders with certain spacing, miniature cylinder surface is covered with immunomagnetic beads.Chipset tumour cell screening, separation/enrichment and detection function are in one, while the chip has dual separation/concentration effect, improve the efficiency of tumour cell detection.
Description
Technical field
The utility model is related to medical instruments fields, and in particular to a kind of micro-fluidic tumour based on immunomagnetic isolation is thin
Born of the same parents' detection chip.
Background technology
In recent years, with micro-fluidic and micromachining technology constantly improve and development, microfluidic chip technology is thin
Advantage in the fields such as biological study and clinical trial diagnosis in born of the same parents' level is increasingly notable.Micro-fluidic chip is also known as chip
Laboratory, the technology are by basic operations such as sample preparation, reaction, separation, detections involved in the fields such as biological and chemical
Unit is integrated or is integrated in substantially on one piece several square centimeters of chip, and network is formed by microchannel, with controlled fluid through whole
A system.The essential characteristic and sharpest edges of the technology are flexible combination of a variety of monotechnicses on small platform and extensive
It is integrated, can by existing all cell analysis step and process, as cell operation, Cell capture/screening, cell culture and
Line real-time dynamic monitoring is analyzed, and is had using particle control chip technology for the detection of tumour cell relative to traditional sensing techniques
There are higher sensitivity and flexibility, highly shortened the Diagnostic Time of tumour.
Development however, as microfluidic chip technology is still within the starting stage at home, relative to the micro-fluidic of foreign countries
Chip detection technique still has the problem of detection sensitivity is relatively low, since tumour cell and immunomagnetic beads reaction time are too short,
Immunomagnetic beads is caused to be combined with tumour cell, tumour cell part is caused to be lost in or be difficult to capture, while utilize traditional electricity
It is relatively low that swimming mode detaches tumour cell and be enriched with purity, tumour cell is caused to be difficult to detect by coming, so as to cause mistaken diagnosis
The problem of.
Utility model content
The utility model to solve the above-mentioned problems, devises a kind of micro-fluidic tumour cell based on immunomagnetic isolation
Detection chip, chipset tumour cell screening, separation/enrichment and detection function are in one, while chip design is with double
Separation/concentration effect again improves the efficiency of tumour cell detection, and the miniature cylinder modified using immunomagnetic beads, which is oriented, catches
The detection sensitivity for improving tumour cell is caught, and the tumour cell of higher degree and the combination of immunoglobulin can be obtained,
Combination detach/be enriched with using galvanomagnetic-effect, improves the detection efficiency and diagnosis rate of tumour cell.
In order to realize above-mentioned technical purpose, reach above-mentioned technique effect, the utility model is real by the following technical programs
Existing：
Micro-fluidic tumour cell detection chip based on immunomagnetic isolation, including chip body, chip body includes base
Piece and optical cement layer composition, it is characterised in that：The optical cement layer is equipped with sample introduction tank, reagent trough a, reagent trough b and waste liquid tank, optical cement
The side of layer is equipped with electrode terminal a and electrode terminal b, and sample introduction tank is connected with reagent trough a by microchannel, behind microchannel
Mixing microchannel is connect, mixing microchannel is followed by separation microchannel, and separation microchannel is followed by dyeing counting microchannel, and the mixing is micro-
Respectively with microchannel and detaching microchannel connection after channel parallel connection, the separation microchannel is detour type microchannel, is detached micro- logical
Magnetic coil is embedded with below road, the both ends of magnetic coil are connect respectively with electrode terminal a and electrode terminal b, mixing
Micro-pillar array is equipped in the channel of microchannel, separation microchannel and dyeing counting microchannel, micro-pillar array is by several miniature cylinders
It is arranged with certain spacing, miniature cylinder surface is covered with immunomagnetic beads.
Preferably, the micro-pillar array is arranged by the miniature cylinder that 80 μm high, spacing is 80 μm.
Preferably, the channel depth of the microchannel, mixing microchannel, separation microchannel and dyeing counting microchannel is
80-200μm。
Preferably, the mixing microchannel is equipped with 4 and is linear type microchannel.
Preferably, the substrate is made of quartz glass, and the optical cement layer is made of dimethyl silicone polymer.
The beneficial effects of the utility model are：Chipset tumour cell screening, separation/enrichment and detection function are in one
Body, while the chip has dual separation/concentration effect, improves the efficiency of tumour cell detection, is modified using immunomagnetic beads
Miniature cylinder be oriented to capture and improve the detection sensitivity of tumour cell, and can obtain the tumour cell of higher degree with
The combination of immunoglobulin to combination detach/be enriched with, improves the detection efficiency of tumour cell using galvanomagnetic-effect
And diagnosis rate.
Description of the drawings
In order to illustrate more clearly of the technical solution of the utility model embodiment, make required for being described below to embodiment
Attached drawing is briefly described, it should be apparent that, the accompanying drawings in the following description is only some embodiments of the utility model,
For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings
Other attached drawings.
Fig. 1 is the Facad structure view of the micro-fluidic tumour cell detection chip based on immunomagnetic isolation.
Fig. 2 is the Section A-A sectional view of the micro-fluidic tumour cell detection chip based on immunomagnetic isolation；
Fig. 3 is the section B-B sectional view of the micro-fluidic tumour cell detection chip based on immunomagnetic isolation.
In attached drawing, parts list represented by the reference numerals are as follows：
1- chip bodies, 101- substrates, 102- optical cement layers, 11- sample introduction tanks, 12- reagent troughs a, 13- reagent trough b, 14- give up
Liquid bath, 16- electrode terminals a, 17- electrode terminal b, 18- magnetic coil, 2- mixing microchannel, 3- separation microchannel, 4-
Dyeing counting microchannel, 5- miniature cylinders.
Specific embodiment
The following is a combination of the drawings in the embodiments of the present utility model, and the technical scheme in the embodiment of the utility model is carried out
It clearly and completely describes, it is clear that described embodiment is only 4 the utility model part of the embodiment rather than whole
Embodiment.Based on the embodiment in the utility model, those of ordinary skill in the art are not making creative work premise
Lower obtained all other embodiment, shall fall within the protection scope of the present invention.
Refering to shown in Fig. 1-3, the micro-fluidic tumour cell detection chip based on immunomagnetic isolation, including chip body
(1), chip body (1) includes optical cement layer made of the substrate (101) made of the quartz glass and dimethyl silicone polymer
(102) it forms, the optical cement layer is equipped with sample introduction tank (11), reagent trough a (12), reagent trough b (13) and waste liquid tank (14), optical cement
The side of layer (102) is equipped with electrode terminal a (16) and electrode terminal b (17), and sample introduction tank (11) and reagent trough a (12) pass through
Microchannel connects, and microchannel is followed by mixing microchannel (2), and mixing microchannel (2) is equipped with 4 and is linear type microchannel, mixes
Microchannel (2) is followed by separation microchannel (3), and separation microchannel (3) is followed by dyeing counting microchannel (4), the mixing microchannel
(2) respectively with microchannel and detaching microchannel (3) connection after in parallel, separation microchannel (3) is detour type microchannel, is divided
From being embedded with magnetic coil (18) below microchannel (3), the both ends of magnetic coil (18) respectively with electrode terminal a (16)
With electrode terminal b (17) connections.
Illustrate the use principle of this chip in the present embodiment for the separation detection of liver cancer cells, detached using this chip
Before detecting liver cancer cells, biotinylation asialoglycoprotein fetuin (ASGPR) is configured and has been coated with magnetic using the immunoglobulin
Microballon adaptive immune magnetic bead, while using the surface of immunomagnetic beads covering miniature cylinder (5), in order to verify the efficiency of covering, make
Surface reaction is verified with biotinylated Cy5-IgG, after Streptavidin has been modified in chip surface reaction, is added
Enter biotin-IgG-Cy5, if chip modification reaction is complete, show that ASGPR is fixed in the miniature cylinder in channel (5).
In order to which the cell in each channel and fixed protein reagent is made to have maximum contact and breaks chip body (1)
The laminar flow of internal flow, this chip is in the channel for mixing microchannel (2), separation microchannel (3) and dyeing counting microchannel (4)
Equipped with micro-pillar array, micro-pillar array is arranged by the miniature cylinder (5) that 80 μm high, spacing is 80 μm, is set by this method
Miniature cylinder, which can not only break laminar flow of fluid, can also prevent cell from rupturing the tumour cell for causing to obtain caused by shock with being immunized
The problem of combination purifying rate of globulin is too low.
Use 1% (w/v) BSA and 0.09% (w/v) NaN3PBS buffer solutions with the upper foster speed of 20 μ L/min
Chip interior is respectively cleaned 3 minutes for channel, loading uses mobile sampling pump by whole blood and immunomagnetic beads containing liver cancer cells
It is added separately in sample introduction tank (11) and reagent trough a (12), whole blood and immunomagnetic beads by branching to 4 respectively behind microchannel
It mixes in microchannel (2), one is equipped with due to being equipped between miniature cylinder (5) and miniature cylinder in every mixing microchannel (2)
Determine Separation, the smaller red blood cell of diameter and leucocyte smoothly run through gap and flow into waste liquid tank (14), reject
After red blood cell and leucocyte, the liver cancer cells in solution are then easier by immunomagnetic beads to capture and combine to form combination；
The liver cancer cells and combination that part does not capture are further flowed in separation microchannel (3), are detached miniature in microchannel
The coated ASGPR of periphery continues to capture the liver cancer cells in mixed solution, since separation microchannel (3) is using circuitous
The channel that declines is returned, greatly extends and captures path, improves the catch rate of tumour cell；Simultaneously by electrode terminal a and electrode
Binding post b is respectively connected to the positive and negative anodes of power supply, and under electromagnetic induction, magnetic coil (18) generates magnetic field, and it is micro- logical to flow into separation
The combination in road (3) is detained under magnetic attraction with detaching in microchannel (3), and the other materials in mixed solution are then smoothly led to
Cross separation microchannel (3)；When mixed solution separation is complete, the power supply of magnetic coil (18) is disconnected, is rinsed using PBS buffer solution
The tumour cell combination in miniature cylinder (5) and separation microchannel (3) is captured to complete the enrichment of tumour cell, enrichment and
The tumour cell of purifying enters in dyeing counting microchannel (4), is injected by reagent trough b (13) into tumour cell combination
Hep Par1 antibody carries out immunofluorescence dyeing, and the tumour cell combination after dyeing is useless in dyeing counting microchannel (4) flow direction
By the Image Acquisition of fluorescence microscope and counting during liquid bath (14).
Using Olympus X70 fluorescence microscope cells in the present embodiment, while using Image-Pro Plus
6.0 softwares carry out Image Acquisition, carefully analyze the image of capture and counting meets the cell of preassigned, color when cell,
The standard compliant cell of the morphological features such as size, core size is identified a liver cancer cells, and then make a definite diagnosis.
In the description of this specification, the description of reference term " one embodiment ", " example ", " specific example " etc. means
At least one of the utility model is contained in reference to the embodiment or example particular features, structures, materials, or characteristics described
In embodiment or example.In the present specification, schematic expression of the above terms be not necessarily referring to identical embodiment or
Example.
The preferred embodiment in the utility model disclosed above is only intended to help to illustrate the utility model.Preferred embodiment is simultaneously
There is no the details that detailed descriptionthe is all, it is only the specific embodiment also not limit the utility model.Obviously, according to this theory
The content of bright book can make many modifications and variations.The utility model is only by claims and its four corner and equivalent
Limitation.
Claims (5)
1. the micro-fluidic tumour cell detection chip based on immunomagnetic isolation, including chip body, chip body includes substrate
It is formed with optical cement layer, it is characterised in that：The optical cement layer is equipped with sample introduction tank, reagent trough a, reagent trough b and waste liquid tank, optical cement layer
Side be equipped with electrode terminal a and electrode terminal b, sample introduction tank connected with reagent trough a by microchannel, and microchannel is followed by mixing
Microchannel is closed, mixing microchannel is followed by separation microchannel, and separation microchannel is followed by dyeing counting microchannel, the mixing microchannel
After parallel connection respectively with microchannel and detach microchannel connection, it is described separation microchannel be detour type microchannel, detach microchannel
Lower section is embedded with magnetic coil, and the both ends of magnetic coil are connect respectively with electrode terminal a and electrode terminal b, mixes micro- logical
Micro-pillar array is equipped in the channel in road, separation microchannel and dyeing counting microchannel, micro-pillar array is by several miniature cylinders with one
Fixed spacing arranges, and miniature cylinder surface is covered with immunomagnetic beads.
2. the micro-fluidic tumour cell detection chip according to claim 1 based on immunomagnetic isolation, it is characterised in that：
The micro-pillar array is arranged by the miniature cylinder that 80 μm high, spacing is 80 μm.
3. the micro-fluidic tumour cell detection chip according to claim 1 based on immunomagnetic isolation, it is characterised in that：
The microchannel, mixing microchannel, separation microchannel and dyeing counting microchannel channel depth be 80-200 μm.
4. the micro-fluidic tumour cell detection chip according to claim 1 based on immunomagnetic isolation, it is characterised in that：
The mixing microchannel is equipped with 4 and is linear type microchannel.
5. the micro-fluidic tumour cell detection chip according to claim 1 based on immunomagnetic isolation, it is characterised in that：
The substrate is made of quartz glass, and the optical cement layer is made of dimethyl silicone polymer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721465011.2U CN207596861U (en) | 2017-11-06 | 2017-11-06 | Micro-fluidic tumour cell detection chip based on immunomagnetic isolation |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721465011.2U CN207596861U (en) | 2017-11-06 | 2017-11-06 | Micro-fluidic tumour cell detection chip based on immunomagnetic isolation |
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| CN207596861U true CN207596861U (en) | 2018-07-10 |
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| CN201721465011.2U Expired - Fee Related CN207596861U (en) | 2017-11-06 | 2017-11-06 | Micro-fluidic tumour cell detection chip based on immunomagnetic isolation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112774743A (en) * | 2019-11-07 | 2021-05-11 | 北京机械设备研究所 | Micro-fluidic chip for enriching cells |
| TWI799879B (en) * | 2020-09-30 | 2023-04-21 | 富佳生技股份有限公司 | Detection chip nucleic acid detection disc and nucleic acid detection |
-
2017
- 2017-11-06 CN CN201721465011.2U patent/CN207596861U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112774743A (en) * | 2019-11-07 | 2021-05-11 | 北京机械设备研究所 | Micro-fluidic chip for enriching cells |
| CN112774743B (en) * | 2019-11-07 | 2022-07-08 | 北京机械设备研究所 | Micro-fluidic chip for enriching cells |
| TWI799879B (en) * | 2020-09-30 | 2023-04-21 | 富佳生技股份有限公司 | Detection chip nucleic acid detection disc and nucleic acid detection |
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| GR01 | Patent grant | ||
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Granted publication date: 20180710 Termination date: 20181106 |