CN108395975A - Unicellular sorting microfluidic device - Google Patents
Unicellular sorting microfluidic device Download PDFInfo
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- CN108395975A CN108395975A CN201710064170.XA CN201710064170A CN108395975A CN 108395975 A CN108395975 A CN 108395975A CN 201710064170 A CN201710064170 A CN 201710064170A CN 108395975 A CN108395975 A CN 108395975A
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- unicellular
- microfluidic device
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- capture
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
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- Dispersion Chemistry (AREA)
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Abstract
The present invention relates to a kind of unicellular sorting microfluidic devices, the microfluidic device of special designing i.e. in flow cytometric sorter, make it is unicellular by the device after, it is sorted into different outlets, so that the sorting work needed for unicellular sequencing becomes more succinct efficient, the technical barrier that difficult label difficulty is sorted in the current unicellular sequencing of effective solution is appropriate for large-scale unicellular sequencing and tests.
Description
Technical field
The present invention relates to biotechnology, especially a kind of unicellular sorting microfluidic device.
Background technology
Gene sequencing technology is a kind of technology being detected to the intragentic collating sequence of organism.Since last century 90
At the beginning of age, the mankind just start to set foot in " Human Genome Project ", are detected by the base composition different to gene in human body
A lot of sequence is obtained, then sequence is analyzed by bioinformatics technique, significant information is obtained, finally helps people
Diagnosed, scientific research, a variety of critical functions such as disease forecasting.
Gene sequencing technology also as other technologies make rapid progress towards faster, more cheap, more accurate side
To development.With the development of sequencing technologies and the significantly decline of sequencing cost, the base from single celled 3,000,000,000 base is decoded
Cell compares sequence and is going to reality because of group and one by one, be push the exact scale of Bioexperiment to unicellular rank, and
It is unconventional to be tested with the tissue containing cells up to a million.
Currently, single celled research achieves preliminary achievement in fields such as proteomics, micro-imagings, grind
Study carefully and finds single celled reaction pattern and non-traditional " the linear result " studied by bulk tissue, but non-zero i.e. 1 number
The result of word.Especially in tumor research field, unicellular research gradually develops into priority fields of study.
Although the technological progress of sequencing instrument makes unicellular sequencing become possible to, unicellular research is still faced with
Very big difficulty.It is each mRNA sequence how marked in individual cells and individual cells.Method general at present
It is that individual cells are sorted into tissue culture plate by flow cytometer（Such as 96 orifice plates）, then pass through polymerase chain reaction technique
The primer of distinguished sequence is added in gene order, carries out single celled positioning by a variety of sequences arranged anyhow, then carry out
Sequencing.But the precision of flow cytometer is relatively low, sorting success rate according to machine state floats between 60 ~ 80%, thin in sorting
Deviation is easy tod produce when born of the same parents, causes experiment invalid, and can only sort 96 unicellular, uncomfortable and mass cell sortings every time
Work.
Invention content
In view of the above-mentioned problems, the present invention provides a kind of alternative solution of the unicellular sorting of flow cytometer, with being completely independent
And simple and easy to do reusable microfluidic device, to substitute the unicellular sorting function of flow cytometer, to improve cell
The accuracy of sorting, the invention is realized in this way：
A kind of unicellular sorting microfluidic device（The size of whole device is about 10x10cm）, which is double-layer structure, under
Layer is the reative cell of liquid communication, and upper layer is inflation air flue control layer；
The reative cell is equipped with several capture chambers being distributed in pairs with array way, each captures there are one being set in chamber " recessed "
Type captures slot, captures up and down between chamber by ganging up pipe unicom, and left and right captures between chamber through cross over pipe unicom, gangs up pipe
It is equipped with air-lock with cross over pipe and switchs mouth.
Further, in the unicellular sorting microfluidic device of flow cytometer of the present invention, 3 are equipped between capture chamber
Volleyball Association's siphunculus.
Further, in the unicellular sorting microfluidic device of flow cytometer of the present invention, each length for capturing chamber
Wide high respectively 90x90x20um, the length, width and height for capturing slot are respectively 30x20x20um, and the length, width and height for capturing notch are respectively
10x10x20um；The length, width and height of communicating pipe are respectively 20x5x20um；The length, width and height of tandem tube are respectively 30x30x20um.
In unicellular screening, individual cells are sorted by microfluidic device provided by the invention by flow cytometer
In, by the unique design of microfluidic device, coordinate label pearl, accurate receipt target cell, sorting success rate up to 80% with
On, it is suitable for extensive unicellular screening.
Description of the drawings
Fig. 1 is reaction chamber structure schematic diagram；
In figure, 1, capture chamber；2, slot is captured；3, notch is captured；4, cross over pipe；5, pipe is ganged up；6, air-lock switchs mouth；7, single
Cell.
Specific implementation mode
A kind of unicellular sorting microfluidic device of flow cytometer, the device are double-layer structure, and lower layer is liquid communication
Reative cell, upper layer be inflation air flue control layer；
Reaction chamber structure captures chambers 1 as shown in Figure 1, it is equipped with two pairs with array way, each captures there are one being set in chamber 1
" recessed " type captures slot 2, is led between the capture chamber 1 of the adjacent two rows in left and right by ganging up 5 unicom of pipe between capture chamber 1 up and down
4 unicom of cross over pipe is crossed, pipe 5 is ganged up and cross over pipe 4 is equipped with air-lock switch mouth 6.
In the present embodiment, left and right is equipped with 3 Volleyball Association's siphunculus 4 between capturing chamber 1.
In the present embodiment, each length, width and height for capturing chamber 1 are respectively 90x90x20um, capture the length, width and height difference of slot 2
For 30x20x20um, the length, width and height for capturing notch 3 are respectively 10x10x20um；The length, width and height of communicating pipe 4 are respectively
20x5x20um；The length, width and height of tandem tube 5 are respectively 30x30x20um.
In the present embodiment, " recessed " type capture slot 2 act as capturing individual cells, is to be caught in capture notch 3
Individual cells 7.
Application method：
1. opening gangs up pipe 5, cross over pipe 4 is closed.
2. respectively to injection cell suspension and label pearl suspension in chamber 1 is captured, it is made to capture individual cells and single
Pearl.
3. pipe 5 is ganged up in closing, cross over pipe 4 is opened, the chamber of lysate and label pearl is housed and single celled chamber is housed
Mixing, cell is cleaved, and mRNA dissociates to being combined with distinguished sequence on label pearl.
4. closing cross over pipe 4, opening gangs up pipe 5, and label pearl is collected in reversed release, real for further handling and being sequenced
It tests.
Above-mentioned specific implementation mode and experiment, its purpose is that description in detail is made to the present invention, but on the basis of the present invention,
It can be made some modifications or improvements, this will be apparent to those skilled in the art, therefore, without departing from this hair
These modifications or improvements on the basis of bright spirit, belong to the scope of protection of present invention.
Claims (2)
1. a kind of unicellular sorting microfluidic device, which is characterized in that the microfluidic device is double-layer structure, and lower layer is liquid
The reative cell of body circulation, upper layer are inflation air flue control layer；
The reative cell is equipped with several capture chambers being distributed in pairs with array way, captures and is equipped with the capture of " recessed " type in chamber
Slot controls between capturing chamber through cross over pipe unicom by ganging up pipe unicom between capture chamber up and down, gangs up pipe and unicom
Pipe is equipped with air-lock switch mouth.
2. unicellular sorting microfluidic device as described in claim 1, which is characterized in that be equipped with 3 Volleyball Associations between capture chamber
Siphunculus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710064170.XA CN108395975A (en) | 2017-02-04 | 2017-02-04 | Unicellular sorting microfluidic device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710064170.XA CN108395975A (en) | 2017-02-04 | 2017-02-04 | Unicellular sorting microfluidic device |
Publications (1)
| Publication Number | Publication Date |
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| CN108395975A true CN108395975A (en) | 2018-08-14 |
Family
ID=63093413
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| Application Number | Title | Priority Date | Filing Date |
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| CN201710064170.XA Pending CN108395975A (en) | 2017-02-04 | 2017-02-04 | Unicellular sorting microfluidic device |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109251841A (en) * | 2018-08-21 | 2019-01-22 | 中国科学院上海微系统与信息技术研究所 | The unicellular sorting chip of one kind and its manufacturing method and unicellular method for separating |
| CN109920482A (en) * | 2019-01-29 | 2019-06-21 | 厦门大学 | A method of analyzing unicellular content |
| WO2021215996A1 (en) * | 2020-04-24 | 2021-10-28 | Lauschke Volker Martin | Device |
-
2017
- 2017-02-04 CN CN201710064170.XA patent/CN108395975A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109251841A (en) * | 2018-08-21 | 2019-01-22 | 中国科学院上海微系统与信息技术研究所 | The unicellular sorting chip of one kind and its manufacturing method and unicellular method for separating |
| CN109251841B (en) * | 2018-08-21 | 2021-09-10 | 中国科学院上海微系统与信息技术研究所 | Single cell sorting chip, manufacturing method thereof and single cell sorting method |
| CN109920482A (en) * | 2019-01-29 | 2019-06-21 | 厦门大学 | A method of analyzing unicellular content |
| WO2021215996A1 (en) * | 2020-04-24 | 2021-10-28 | Lauschke Volker Martin | Device |
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Legal Events
| Date | Code | Title | Description |
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| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180814 |
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| WD01 | Invention patent application deemed withdrawn after publication |