CN105699671A - Small micro-fluidic chip system for biological particle parting analyzing - Google Patents
Small micro-fluidic chip system for biological particle parting analyzing Download PDFInfo
- Publication number
- CN105699671A CN105699671A CN201610031688.9A CN201610031688A CN105699671A CN 105699671 A CN105699671 A CN 105699671A CN 201610031688 A CN201610031688 A CN 201610031688A CN 105699671 A CN105699671 A CN 105699671A
- Authority
- CN
- China
- Prior art keywords
- lenticule
- micro
- biological particle
- fluidic chip
- load
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention relates to a small micro-fluidic chip system for biological particle parting analyzing and belongs to the field of biological sample detection.The small micro-fluidic chip system comprises an excitation fiber channel, an excitation slice carrying micro lens, a collection slice carrying micro lens and a collection fiber channel.The excitation slice carrying micro lens and the collection slice carrying micro lens are arranged on the two sides of a waste liquid microchannel respectively.The optical axis of the excitation slice carrying micro lens is perpendicular to the waste liquid microchannel.An included angle is formed between the optical axis of the collection slice carrying micro lens and the waste liquid channel.The excitation fiber channel and the collection fiber channel are formed in a chip.The micro-fluidic chip system is simple in structure, small in size, low in cost and power consumption, high in sensitivity, convenient to operate and free of optical calibration, and detection is accurate.A plurality of slice carrying micro lenses-fiber detection modules on the micro-fluidic chip can be machined to work in parallel, which helps to achieve high-flux detection.Therefore, the system is suitable for analyzing detection of a plurality of kinds of biological samples, especially for biological particles.
Description
Technical field
The present invention relates to a kind of small-sized micro-fluidic chip system for biological particle phenotypic analysis, belong to biology sample detection field。
Background technology
Flow cytometry is a kind of phenotypic analysis conventional sense technology for biological particle, is widely used in the fields such as clinical diagnosis, biochemistry, biology and environmental monitoring。Although Fluorescein activated cell sorter is very advanced, but traditional stream type cell analyzer volume is big, power consumption is high, expensive, operate relatively complicated, under some extreme environments and condition, these equipment cannot normally play a role, in emergency relief, space station, frontier sentry, remote country etc.。Micro-total analysis system (miniaturizedtotalanalysissystem, μ-TAS) also referred to as chip lab (Laboratory-on-a-Chip, it is called for short LOC) it is proposed by Manz and the Widmer of CibaGeigy company of Switzerland first nineteen ninety, obtain swift and violent development afterwards。Development through decades, the many advantages such as micro-fluidic chip is quick by it, high flux, low-power consumption, portability, being easily integrated, being widely used in the phenotypic analysis research of biological particle, the miniaturization for biological particulate instrument device proposes new thinking。The DavidHolmes etc. of Southampton, Britain university proposes a kind of Integrated electrode off-chip detection micro-fluidic flow cytometry number system of biological particle fluorescence signal on micro-fluidic chip, achieve measuring (HolmesD. while biological particle optics and electrical information, PettigrewD., RecciusC.H.etal.LeukocyteAnalysisandDifferentiationUsing HighSpeedMicrofluidicSingleCellImpedanceCytometry [J] .LabOnaChip, 2009,9 (20): 2881-2889.)。As shown in Figure 1, although the volume of micro-fluidic chip is only small, but the auxiliary equipment that a pile is complicated is had, such as laser instrument, detector, liquid driving device and electronics, make final device volume still quite huge, and the integration of space optics parts and micro-fluidic chip and calibration difficulties, consequent power consumption and instrument cost are still higher, the advantage completely micro-fluidic chip brought balances out, sample pretreatment step is made to become loaded down with trivial details on the contrary, counting precision and detection sensitivity are all barely satisfactory, but without the low cost realizing real meaning, miniaturization, rapid and portability。
Therefore, this patent, mainly for the technical characterstic of micro-fluidic chip, has been invented a kind of miniaturization, low cost, low-power consumption, has been prone to the micro flow control chip device that biological particle can carry out multiparameter detection integrated with micro-fluidic chip, simple to operate, without optical correction。
Summary of the invention
The invention aims to solve that prior art volume is big, complicated operation and the problem difficult integrated with micro-fluidic chip, it is provided that a kind of small-sized micro-fluidic chip system for biological particle phenotypic analysis。
It is an object of the invention to be achieved through the following technical solutions。
A kind of small-sized micro-fluidic chip system for biological particle phenotypic analysis, including sheath fluid microchannel, sample microchannel, waste liquid microchannel；Also include excitation fiber passage, excite sheet load lenticule, collecting tab load lenticule, collect optical-fibre channel；Described sheet load lenticule and collecting tab load lenticule is excited to be respectively placed in both sides, waste liquid microchannel；Excite sheet to carry lenticular optical axis to be mutually perpendicular to waste liquid microchannel；Collecting tab is carried lenticular optical axis and is had angle with waste liquid microchannel, need to ensure that the angle of this angle is not 90 °；Excitation fiber passage is opened on chip with collecting optical-fibre channel, and wherein excitation fiber passage carries lenticular light shaft coaxle with exciting sheet, collects optical-fibre channel and carries lenticular light shaft coaxle with collecting tab。
Work process: be in that to utilize MEMS technology to process the micro flow control chip device that biological particle can carry out multiparameter detection, use it for the phenotypic analysis of various biological particle。This micro-fluidic chip system adopts two dimension folder circulation road sample introduction, and biological particle sample, under the parcel of sheath fluid, forms a line and passes through detection zone one by one。Laser is through fiber coupler entrance excitation fiber to micro-fluidic chip, formed through exciting sheet load lenticule to entreat in the channel and detect the hot spot that biological particle diameter dimension is close, when single detection biological particle is by detection zone, produce forward scattering light, side scattered light and fluorescence, and carried lenticule by the collecting tab of passage offside respectively and collect optical fiber and collect to photodetector。Through data collecting card, the data collected are reached PC and be analyzed display。
Beneficial effect
The micro-fluidic chip system simple in construction that uses in the present invention, volume is little, low cost, low-power consumption and this system sensitivity is high, it is accurate, easy to operate to detect, without optical correction, and the sheet load lenticule-fiber detection module on micro-fluidic chip can process several concurrent workings, contributes to realizing high throughput testing。Therefore, native system detects suitable in the analysis of multiple biological sample especially biological particle。
Accompanying drawing explanation
Fig. 1 is the system construction drawing that document " LeukocyteAnalysisandDifferentiationUsingHighSpeedMicrofl uidicSingleCellImpedanceCytometry " proposes；
Fig. 2 be the present invention specifically assemble structural representation；
Fig. 3 is micro-fluidic chip concrete structure schematic diagram；
Fig. 4 is the forward-scattering signal collection result of 10 μ m diameter microspheres。
Wherein, 1 sheath fluid microchannel, 2 sample microchannels, 3 waste liquid microchannels, 4 excitation fiber passages, 5 excite sheet load lenticule, 6 collecting tab load lenticulees, 7 collect optical-fibre channels。
Detailed description of the invention
Below in conjunction with accompanying drawing, the invention will be further described with embodiment。
Embodiment 1
A kind of small-sized micro-fluidic chip system for biological particle phenotypic analysis, as it is shown on figure 3, include sheath fluid microchannel 1, sample microchannel 2, waste liquid microchannel 3；Also include excitation fiber passage 4, excite sheet load lenticule 5, collecting tab load lenticule 6, collect optical-fibre channel 7；Described sheet load lenticule 5 is excited to be respectively placed in both sides, waste liquid microchannel 3 with collecting tab load lenticule 6；The optical axis exciting sheet load lenticule 5 is mutually perpendicular to waste liquid microchannel 3；The optical axis of collecting tab load lenticule 6 has angle with waste liquid microchannel 3, need to ensure that the angle of this angle is not 90 °；Excitation fiber passage 4 is opened on chip with collecting optical-fibre channel 7, and wherein excitation fiber passage 4 and the light shaft coaxle exciting sheet load lenticule 5, collect the light shaft coaxle of optical-fibre channel 7 and collecting tab load lenticule 6。
1, substrate designs
Pass through detection zone one by one to allow biological particle form a line in going out circulation road, it is achieved single microgranule sample introduction, utilize fluid mechanical emulation software that microfluidic channel structure, inlet flow rate have been carried out design of Simulation。In order to 633nm laser is introduced micro flow control chip device, and in going out circulation road, form the detection hot spot close with particle size, utilize optical simulation software to simulate from single-mode fiber (4 μm of core diameter 0.13NA) end face outgoing beam transmission path on micro-fluidic chip, designed and produced and excited sheet load lenticule and collecting tab load lenticule。
2, substrate makes
[1] mask design
Utilize mapping software design photo etched mask, microchannel, optical-fibre channel and sheet are carried microlens structure and designs on mask, export mask film with laser photocomposing machine。
[2] whirl coating
SU-8 photoresist spin coating amount is 1ml/inch, adopts 3 inch silicon wafer, therefore falls 3mlSU-8 photoresist in wafer center, whirl coating is carried out: 500r/min spin coating 10s with spin coating all-in-one, 1375r/min spin coating 30s, stands elimination edge effect in 10 minutes and obtains the glue surface of 130 μ m-thick。
[3] pre-baked: pre-baked purpose is to make the solvent in SU-8 photoresist volatilize, and can take away the tiny bubble in photoresist, therefore rate controlled is crucial。Being risen to 95 DEG C with the speed of 5 DEG C/min by room temperature on heating platform in experiment, period keeps 5min and 25min respectively at 65 DEG C and 95 DEG C, and slow cooling is to less than 75 DEG C afterwards。
[4] photoetching: the theoretical formula of time of exposure is: time of exposure=exposure dose/exposure machine power, laboratory mercury lamp luminous power is 20mW/cm2, and the glue exposure dose of 130 μ m-thick is between 350mJ/cm2～550mJ/cm2, experiment adopts 450mJ/cm2, and therefore time of exposure is 32.5s。
[5] bake after exposing: owing to SU-8 is negative photoresist, it is therefore desirable to utilize to bake after exposing and reinforce light part bond, in order to avoid figure is dissolved when development。Being risen to 95 DEG C with the speed of 5 DEG C/min by room temperature on heating platform in experiment, period keeps 5min and 12min respectively at 65 DEG C and 95 DEG C, is slowly dropped to room temperature afterwards。
[6] development is with fixing: inserted by silicon chip in developer solution, and concussion makes development more abundant gently simultaneously, rinses with isopropanol or soaks fixing, then clean with plasma water after development, and nitrogen dries up。If discovery silicon chip still remaining white spot, then it represents that underdevelop, it is necessary to repeat development and fixing。
[7] hard baking: be placed on flat plate heat by silicon chip at 120 DEG C of hard baking 10min, makes photoresist firm can be attached to silicon chip surface, to ensure the recycling rate of waterused of mould。
[8] silanization: peel off for making PDMS be prone to from mould, by mould silanization 1h, to complete whole Mold Making。
[9] molding: be placed on horizontal aluminium sheet by clean silicon chip, around parcel one circle adhesive tape, to prevent PDMS from spilling, control PDMS height as fence on the other hand, on the SU-8 mould being cast in by the PDMS having taken off gas, stands 5min and makes it from smooth。Thereafter it is transferred in 100 DEG C of vacuum drying ovens, solidifies 1h。
[10] bonding: after cooling, PDMS structure is released from SU-8 mould, and in outlet, the punching of injection port place, then, it is placed in oxygen plasma surface treatment instrument together with cleaned PDMS/ glass/PMMA cover plate 2, surface treatment 30s, fits the two rapidly after taking-up, form the microchannel (noting extruding bubble) closed, complete chip manufacturing。
3, assemble,
In order to biological particle sample and sheath fluid are introduced fluid channel, miniflow pump is utilized biological particle sample and sheath fluid to be pumped in fluid channel。The present invention can detect three kinds of parameters of biological particle simultaneously: forward scattering light, side scattered light and fluorescence。The light source that the detection of scattering light and fluorescence is required is provided by same 633nm laser instrument。Load single-mode fiber in excitation fiber passage 4, laser instrument output is caused micro flow control chip device；Collect and load multimode fibre in optical-fibre channel 7 and collect scattering that single biological particle produces through detection zone and use up, and transmission is to photodetector。Centered by micro-fluidic chip, by photoelectric detector and relevant device around chip mount around, the overall structure figure of device such as Fig. 2。
4, systematic function
Miniflow pump is used to be pumped in sample microchannel 1 and sheath fluid microchannel 2 respectively by polystyrene microsphere sample and the sheath fluid of 10 μ m diameter, single biological particle sample introduction is realized by controlling the sample introduction speed ratio of sample feeding mouth and sheath fluid injection port place, laser is caused micro-fluidic chip by the excitation fiber in excitation fiber passage 4, through exciting sheet load lenticule 5 to converge to the central authorities of waste liquid microchannel 3, form the detection hot spot close with biological particle diameter dimension, when single biological particle produces forward scattering light through detection zone, side scattered light and side fluorescence, scattering light and fluorescence are collected sheet load lenticule 6 and coupled to collection optical fiber and detected by photodetector, thus the multiparameter detection realizing biological particle sample is analyzed。Fig. 4 is the forward-scattering signal of the 10 μ m diameter microspheres utilizing native system to collect, through statistical computation, the coefficient of variation CV of native system is about 18%, Signal-to-Noise SNR is about 20dB, with existing biological particle phenotypic analysis system (CV:20%～30% based on micro-fluidic chip, SNR～9dB) to compare, the homogeneous microsphere of a certain size is had well response concordance and stability by the present invention, has more superior systematic function。
Claims (2)
1. for a small-sized micro-fluidic chip system for biological particle phenotypic analysis, including sheath fluid microchannel (1), sample microchannel (2), waste liquid microchannel (3)；It is characterized in that: also include excitation fiber passage (4), excite sheet load lenticule (5), collecting tab load lenticule (6), collect optical-fibre channel (7)；Described excite sheet load lenticule (5) with collecting tab load lenticule (6) be respectively placed in waste liquid microchannel (3) both sides；The optical axis exciting sheet load lenticule (5) is mutually perpendicular to waste liquid microchannel (3)；The optical axis of collecting tab load lenticule (6) and waste liquid microchannel (3) have angle, and need to ensure that the angle of this angle is not 90 °；Excitation fiber passage (4) is opened on chip with collecting optical-fibre channel (7), wherein excitation fiber passage (4) and the light shaft coaxle exciting sheet load lenticule (5), collect the light shaft coaxle of optical-fibre channel (7) and collecting tab load lenticule (6)。
2. a kind of small-sized micro-fluidic chip system for biological particle phenotypic analysis as claimed in claim 1, it is characterized in that: the work process of described system: be in that to utilize MEMS technology to process the micro flow control chip device that biological particle can carry out multiparameter detection, use it for the phenotypic analysis of various biological particle；This micro-fluidic chip system adopts two dimension folder circulation road sample introduction, and biological particle sample, under the parcel of sheath fluid, forms a line and passes through detection zone one by one；Laser is through fiber coupler entrance excitation fiber to micro-fluidic chip, formed through exciting sheet load lenticule to entreat in the channel and detect the hot spot that biological particle diameter dimension is close, when single detection biological particle is by detection zone, produce forward scattering light, side scattered light and fluorescence, and carried lenticule by the collecting tab of passage offside respectively and collect optical fiber and collect to photodetector；Through data collecting card, the data collected are reached PC and be analyzed display。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610031688.9A CN105699671A (en) | 2016-01-18 | 2016-01-18 | Small micro-fluidic chip system for biological particle parting analyzing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610031688.9A CN105699671A (en) | 2016-01-18 | 2016-01-18 | Small micro-fluidic chip system for biological particle parting analyzing |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105699671A true CN105699671A (en) | 2016-06-22 |
Family
ID=56226356
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610031688.9A Pending CN105699671A (en) | 2016-01-18 | 2016-01-18 | Small micro-fluidic chip system for biological particle parting analyzing |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105699671A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108414477A (en) * | 2018-01-12 | 2018-08-17 | 山东省科学院海洋仪器仪表研究所 | Seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus and method |
| CN111504907A (en) * | 2020-04-09 | 2020-08-07 | 江苏师范大学 | Method for detecting luminescence signal in microchannel |
| CN111889153A (en) * | 2020-08-04 | 2020-11-06 | 桂林电子科技大学 | Flow cytometer based on optical fiber integrated microfluidic chip |
| CN112147044A (en) * | 2020-09-07 | 2020-12-29 | 桂林电子科技大学 | Spectrum subdivision type optical fiber distributed detection device for flow cytometer |
| CN112229781A (en) * | 2020-09-07 | 2021-01-15 | 桂林电子科技大学 | Improved spectrum subdivision type optical fiber distributed detection device of flow cytometer |
| CN113941377A (en) * | 2021-10-11 | 2022-01-18 | 北京理工大学 | All-glass microfluidic chip and processing method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103398936A (en) * | 2009-07-07 | 2013-11-20 | 索尼公司 | Microfluidic device |
| CN103512862A (en) * | 2012-06-15 | 2014-01-15 | 黄辉 | Signal difference detection based optical microfluidic chip and test method |
| CN104136922A (en) * | 2012-02-04 | 2014-11-05 | 细胞和分子平台中心（C-Camp） | Microfluidic flow analyzer and method thereof |
-
2016
- 2016-01-18 CN CN201610031688.9A patent/CN105699671A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103398936A (en) * | 2009-07-07 | 2013-11-20 | 索尼公司 | Microfluidic device |
| CN104136922A (en) * | 2012-02-04 | 2014-11-05 | 细胞和分子平台中心（C-Camp） | Microfluidic flow analyzer and method thereof |
| CN103512862A (en) * | 2012-06-15 | 2014-01-15 | 黄辉 | Signal difference detection based optical microfluidic chip and test method |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108414477A (en) * | 2018-01-12 | 2018-08-17 | 山东省科学院海洋仪器仪表研究所 | Seawater chlorophyll a, phycocyanin and phycoerythrin parameter measuring apparatus and method |
| CN111504907A (en) * | 2020-04-09 | 2020-08-07 | 江苏师范大学 | Method for detecting luminescence signal in microchannel |
| CN111889153A (en) * | 2020-08-04 | 2020-11-06 | 桂林电子科技大学 | Flow cytometer based on optical fiber integrated microfluidic chip |
| CN112147044A (en) * | 2020-09-07 | 2020-12-29 | 桂林电子科技大学 | Spectrum subdivision type optical fiber distributed detection device for flow cytometer |
| CN112229781A (en) * | 2020-09-07 | 2021-01-15 | 桂林电子科技大学 | Improved spectrum subdivision type optical fiber distributed detection device of flow cytometer |
| CN113941377A (en) * | 2021-10-11 | 2022-01-18 | 北京理工大学 | All-glass microfluidic chip and processing method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105699671A (en) | Small micro-fluidic chip system for biological particle parting analyzing | |
| CN103191791B (en) | Integrated chip system for high-throughput sorting and counting detection of biological particles, and application | |
| Rosenauer et al. | Miniaturized flow cytometer with 3D hydrodynamic particle focusing and integrated optical elements applying silicon photodiodes | |
| Chiu et al. | Universally applicable three-dimensional hydrodynamic microfluidic flow focusing | |
| Godin et al. | Microfluidics and photonics for Bio‐System‐on‐a‐Chip: A review of advancements in technology towards a microfluidic flow cytometry chip | |
| US9442106B2 (en) | Simple and affordable method for immunophenotyping using a microfluidic chip sample preparation with image cytometry | |
| CN102527280B (en) | Micro mixing and micro reaction device | |
| US7999937B1 (en) | Microfluidic devices and methods for integrated flow cytometry | |
| CN104931440B (en) | A kind of Portable heavy metal high-sensitivity detection device based on micro-fluidic chip | |
| Suthanthiraraj et al. | One-dimensional acoustic standing waves in rectangular channels for flow cytometry | |
| CN203220910U (en) | Integrated chip for high-throughput sorting and count detection of biological particles | |
| US20050239210A1 (en) | Analytical chip and analytical apparatus | |
| Rosenauer et al. | Characterization of a microflow cytometer with an integrated three-dimensional optofluidic lens system | |
| Zhao et al. | Universally applicable three-dimensional hydrodynamic focusing in a single-layer channel for single cell analysis | |
| Fan et al. | Three dimensional microfluidics with embedded microball lenses for parallel and high throughput multicolor fluorescence detection | |
| Hu et al. | Precise label-free leukocyte subpopulation separation using hybrid acoustic-optical chip | |
| Lien et al. | A prealigned process of integrating optical waveguides with microfluidic devices | |
| Tottori et al. | Particle/cell separation using sheath-free deterministic lateral displacement arrays with inertially focused single straight input | |
| CN105319197A (en) | Liquid drop micro-fluidic chip based on microlens array | |
| Guo et al. | A compact optofluidic cytometer for detection and enumeration of tumor cells | |
| Li et al. | Ultraportable flow cytometer based on an all-glass microfluidic Chip | |
| Fu et al. | Optical microflow cytometer based on external total reflection | |
| TWI257480B (en) | Cellular micro-particle detection chip and manufacturing method thereof | |
| Leary | Design of portable microfluidic cytometry devices for rapid medical diagnostics in the field | |
| CN102507528A (en) | Micro-fluid detection system based on self-focusing lens |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160622 |
|
| WD01 | Invention patent application deemed withdrawn after publication |