CN2821565Y - Single cell algae flow type analysis microflow control chip - Google Patents
Single cell algae flow type analysis microflow control chip Download PDFInfo
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- CN2821565Y CN2821565Y CN 200520084372 CN200520084372U CN2821565Y CN 2821565 Y CN2821565 Y CN 2821565Y CN 200520084372 CN200520084372 CN 200520084372 CN 200520084372 U CN200520084372 U CN 200520084372U CN 2821565 Y CN2821565 Y CN 2821565Y
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Abstract
The utility model discloses a micro-flow control chip for single-cell algae flowing type analysis, which comprises a base chip and a cover chip which have symmetrical structures, wherein a psi type liquid flowing system comprising a middle sample passage and two side sheath flowing channels, and an optical fiber channel for detecting are arranged on the base chip and the cover chip; the sample channel is consistent with the sizes of the sheath flowing channels which form a 1/4 circular-arc shape, and the sample channel is consistent with the tangential directions of the end points of the sheath flowing channels; the optical fiber channel perpendicular to a sample injecting channel is arranged on the micro-flow control chip for single-cell algae flowing type analysis, coupled fibers of a laser are horizontally inserted in the fiber channel, and excitation light is led next to a detecting area of a separating channel; an optical detecting system is arranged above a monitoring area of the sample injecting channel of the micro-flow control chip for single-cell algae flowing type analysis and is used for detecting fluorescence and lateral scattering light, and then the utility model carries out counting and classification for alga through the comprehensive analysis of light signals. The utility model leaves out a complicated light path system using a plurality of optical elements to accomplish three-dimensional collimation, reflection and convergence, reduces the strength loss in the process of the light transmission to the minimum and realizes the characteristics of the miniaturization, the portability and the integration of a micro-flow control chip system.
Description
Technical field
The utility model relates to a kind of micro-fluidic chip that is applicable to the marine unicellular algae detection that utilizes the flow cytometer principle design to make.
Background technology
Micro-total analysis system (Micro-total analysis system, μ-TAS) proposition of notion has produced significant impact in the analysis science field, and the guiding chemical analysis apparatus is towards the trend development of microminiaturized, integrated and portability.It utilizes micro fabrication to make functional units formation very small chemical systems such as little valve, microchannel, microreactor, microflow sensor, little detecting device on chip.Micro-fluidic chip system has efficiently, low consumption, microminiaturization and integrated characteristics, is suitable for all kinds of on-the site analysiss and The real time measure.Nineteen ninety-five Mathies and Woolley adopt little electrophoresis chip to carry out dna sequencing research first, on the passage of effective separation length 3.5cm, and about 150 bases of order-checking in the 10min, accuracy rate 97%.The microchip gene alaysis system of functions such as integrated cell on micro-fluidic chip such as Ramsey in 1998 is cleared up, pcr amplification and electrophoretic separation.
The principle of work of flow cytometer is: will make single cell suspension behind the cell dyeing to be measured, with certain pressure testing sample is pressed into flow chamber, not celliferous phosphate buffer under high pressure sprays from the sheath fluid pipe, sheath fluid tube inlet direction and testing sample stream are angled, like this, sheath fluid just can hold the sample flow at high speed, forms a fluid stream of a circle, cell to be measured single file under the bag quilt of sheath fluid is arranged, and passes through surveyed area successively.
Use flow cytometer to detect miniature phytoplankton and just adopted by more and more researchers, Paau etc. and Yentsch etc. have done correlative study, have promoted the utilization of flow cytometry in the miniature phytoplankton in ocean detects.Flow cytometer usually with laser as light emitting source.Through the light beam after the focusing shaping, vertical irradiation is on sample flow, and pigment and other materials produce scattered light and fluorescence excitation in the cell body under the irradiation of laser beam.The forward angle light scatter light intensity is relevant with the size of cell.The particle properties that the measurement of side scattered light is mainly used to obtain cells involved internal fine structure for information about.The specific wavelength fluorescence signal intensity has been represented the composition and the content of the pigment in the single-cell algae of surveying.But at present, flow cytometer costs an arm and a leg, and complex structure is not suitable for on-the site analysis, and the rig-site utilization that appears as Flow Cytometry of micro-fluidic chip provides may.
Summary of the invention
The purpose of this utility model provides a kind of miniature, portable and be suitable for the micro-fluidic chip of on-the site analysis, and it can be counted and the population composition analysis marine unicellular algae.
The utility model is based on standard photolithography techniques, is the micro-fluidic chip material with glass, makes up microchannel and little liquid storage tank on chip, makes the sample introduction of sample, detection be integrated on the chip and finish.Micro-fluidic chip comprises the substrate and the cover plate of structural symmetry, the optical-fibre channel that is built with liquid fluid system and is used to detect on substrate and cover plate.Liquid fluid system adopts ψ type structure, center-aisle is a sample channel, the subchannel, both sides is called the sheath circulation road, the effect of sheath circulation road is that the frustule in the sample is arranged with single file, sample channel and sheath circulation road consistent size, and the sheath circulation road is 1/4 circular-arc, sample channel is consistent with sheath circulation road end points tangential direction, sheath stream converges in the same way with sample flow like this, avoids liquid stream to converge the generation vortex disturbance, has guaranteed the stability of liquid fluid system; The end of sample channel and sheath circulation road is built with liquid storage tank respectively, and the other end of sample channel also has a waste liquid pool; On chip, make up the optical-fibre channel vertical, make laser instrument coupled fiber level insert this passage, exciting light is incorporated near the surveyed area of split tunnel with sample intake passage.The position parallel with the incident light optical-fibre channel at the opposite side of sample intake passage makes up optical-fibre channel, is used for forward scattering light is introduced the respective detection device.Above chip sample intake passage monitored area, settle the optical detection system, detect fluorescence and side scattered light, when the frustule process detects mouth, be subjected to laser radiation, produce fluorescence (yellow fluorescence, red fluorescence, green fluorescence) and scattered light (forward scattering and lateral scattering), by analysis-by-synthesis, algae is counted and classifies several light signals.This chip has saved three-dimensional collimation, the reflection that just can be finished by a plurality of optical elements and has assembled the complex optical path system, the loss of strength in the optical transmission process is dropped to minimum, has realized microminiaturization, portability and the integrated characteristics of micro-fluidic chip system.
The utility model has embodied the microminiaturization of micro-fluidic chip system and integrated characteristics with the flow cytometer liquid fluid system and the optical detection system integration, is suitable for all kinds of on-the site analysiss and The real time measure.Be widely used in fields such as single-cell algae, cultured cell, blood cell.
Description of drawings
Fig. 1 is the structural representation of micro-fluidic chip of the present utility model.
Wherein, 1-sample inlet; 2,3-sheath inflow entrance; The 4-waste liquid pool; The 5-sample channel; 6,7-sheath circulation road; 8-passage joint; 9-incident optical passage, 10-outgoing optical-fibre channel.
Fig. 2 is the structural representation of single-cell algae flow cytometer showed chip system.
Wherein, 11,12, the 13-micropump; 14,16-sheath stream; The 15-sample; 17,18, the 19-fluorescence detector; 20-lateral scattering photodetector; The 21-bandpass filter; The 22-dichroic filter; The 23-laser instrument, the 24-detecting device.
Embodiment
Further specify the utility model below in conjunction with accompanying drawing and by specific embodiment.
Embodiment 1: the structure of micro-fluidic chip
As shown in Figure 1, microfluidic chip structure divides two parts: liquid fluid system and detection system.Liquid fluid system adopts ψ type structure, center-aisle is a sample channel 5, the subchannel, both sides is sheath circulation road 6 and 7, and the effect of sheath circulation road is to make the frustule in the sample be single file arrangement, sample channel 5 and sheath circulation road 6 and 7 consistent size, the sheath circulation road is 1/4 circular-arc, sheath circulation road 6 and 7 consistent with sample channel 5 directions in the tangential direction of joint 8, like this, sheath stream converges in the same way with sample flow, avoid liquid stream to converge the generation vortex disturbance, guaranteed the stability of liquid fluid system; On chip, make up the optical-fibre channel 9,10 vertical, make laser instrument 23 coupled fiber levels insert this passage, exciting light is incorporated near the surveyed area of split tunnel with sample channel.The position parallel with the incident light optical-fibre channel at the opposite side of sample intake passage makes up optical-fibre channel, is used for forward scattering light is introduced detecting device 24.Above chip sample intake passage surveyed area, settle the optical detection system, fluorescence detector 17,18,19 and lateral scattering photodetector 20 receive the corresponding light signal respectively, when the frustule process detects mouth, be subjected to laser radiation, produce fluorescence (red fluorescence, yellow fluorescence, green fluorescence) and scattered light (forward scattering and lateral scattering), fluorescence and scattered light arrive the respective detection device by the selective transmission and the reflection of dichroic filter 22 and bandpass filter 21.By analysis-by-synthesis, algae is counted and classifies several light signals.This chip has saved three-dimensional collimation, the reflection that just can be finished by a plurality of optical elements and has assembled the complex optical path system, the loss of strength in the optical transmission process is dropped to minimum, has realized microminiaturization, portability and the integrated characteristics of micro-fluidic chip system.
Embodiment 2: the making of micro-fluidic chip
1. the making of glass substrate: the size design of microchannel on the mask film: fluid passage, 30 μ m; Optical-fibre channel, 60 μ m; Between optical-fibre channel and split tunnel apart from d:400 μ m.The mask film is placed on the even glue chromium plate of 63mm * 63mm * 1.5mm, ultraviolet exposure 180 seconds (wavelength 365nm) after developing 100 seconds in the developer solution, is dried half an hour down for 100 ℃.At room temperature use chromium film etching liquid (cerous sulfate: perchloric acid: water=50 grams: 15 milliliters: 300 milliliters) corrode the chromium film, rinse oven dry then with high purity water well.By the digit microscope shooting, the channel size that records on the chromium plate is: fluid passage, 40 μ m; Optical-fibre channel, 70 μ m.With 0.5M HF/0.5M NH
4The exposed Pyrex of F etching agent corrosion, speed is about 10 μ m/h, and etching is removed residual light glue-line and chromium film with acetone, chromium film etching liquid after 6 hours more successively, promptly gets substrate.With miniature bench drill punching, drill bit is the diamond bit of 1mm, and the diameter in hole is liquid pool diameter 1mm.Microscopically is measured the microchannel size: microfluidic channel, last bottom width 160 μ m, following bottom width 50 μ m, the degree of depth 60 μ m; Optical-fibre channel, last bottom width 190 μ m, following bottom width 80 μ m, the degree of depth 60 μ m; Between optical-fibre channel and split tunnel apart from d:340 μ m; Other zones except that optical-fibre channel are protected with adhesive tape, continue the etching optical-fibre channel, reach bottom width 600 μ m up to channel size, following bottom width 80 μ m, the degree of depth 270 μ m; Between optical-fibre channel and split tunnel apart from d:120 μ m
2. the making of cover plate: identical with the substrate method for making, construct identically, but do not punch.
With substrate and cover plate successively at acetone, H
2O-H
2O
2-NH
4OH (5: 1: 1) solution, H
2SO
4: H
2O
2In (4: 1) solution and the high purity water ultrasonic cleaning 5-10 minute, dry up the sealing of in super-clean environment, both being alignd then, bonding under the high temperature with nitrogen.Heating schedule is: rise to 550 ℃ with 40 ℃/min from room temperature, 30 minutes time; Rise to 610 ℃, 30 minutes time with 20 ℃/min from 550 ℃; Rise to 635 ℃, 30 minutes time with 20 ℃/min from 610 ℃; Rise to 650 ℃, 6 hours time with 10 ℃/min from 635 ℃.Naturally cool to room temperature then.Through microscopically observation, passage does not have distortion behind the bonding, and reaches sealing fully.Chip microchannel ovalize behind the bonding, sample channel major axis are about 190 μ m, and minor axis is about 120 μ m; The about 600 μ m of light channel major axis, the about 540 μ m of minor axis.
Embodiment 3: micro-fluidic chip analyzing and testing system
As shown in Figure 2, connect sampling pump 12 in the import 1 of sample channel 5, both sides sheath circulation road 6 and 7 connects sheath stream 14,16 at its inlet 2 and 3 places.Sample and sheath flow liquid are by pump 11,12 and 13 carry, sheath stream and sample co-flow, it is mobile forward with the form of laminar flow that sheath flow liquid and sample converge the back at point 8, by regulating sheath stream and sample flow rate, making cells in sample be single file arranges, and be advanced through detection zone, in of the laser excitation of surveyed area cell quilt from optical fiber, the optical fiber of passage opposite side is accepted forward scattering light, and it is transferred to fluorescence detector 24, the light of side direction is through the beam split of dichroic filter 22 and bandpass filter 21, filter, respectively by lateral scattering photodetector 20, red fluorescence detecting device 17, yellow fluorescence detecting device 18 and green fluorescence detecting device 19 receiving records, comprehensive all kinds of light signals obtain the information of miniature phytoplankton quantity and structure of community.
The utility model has made up on the micro-fluidic chip that utilizes the flow cytometer ultimate principle to design and produce realizes required liquid fluid system and the sense channel of Flow Cytometry, has realized the microminiaturization of single-cell algae flow cytometer showed system, integrated and simplification; Low cost of manufacture is easy to realize standardization and large-scale production.
Claims (4)
1. micro-flow controlling chip for analyzing single cell algae flow, it is characterized in that it comprises the substrate and the cover plate of structural symmetry, on substrate and cover plate, be built with the liquid fluid system of the ψ type structure that comprises middle sample passage and both sides sheath circulation road and the optical-fibre channel that is used to detect.
2. micro-fluidic chip according to claim 1 is characterized in that described sheath circulation road is 1/4 circular-arc, and sample channel is consistent with sheath circulation road go side point tangential direction.
3. micro-fluidic chip according to claim 1 is characterized in that described optical-fibre channel and sample intake passage are orthogonal.
4. micro-fluidic chip according to claim 1 is characterized in that it is material with glass.
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| CN 200520084372 CN2821565Y (en) | 2005-06-19 | 2005-06-19 | Single cell algae flow type analysis microflow control chip |
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| CN 200520084372 CN2821565Y (en) | 2005-06-19 | 2005-06-19 | Single cell algae flow type analysis microflow control chip |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102507528A (en) * | 2011-12-02 | 2012-06-20 | 徐州雷奥医疗设备有限公司 | Micro-fluid detection system based on self-focusing lens |
| CN103234949A (en) * | 2013-01-30 | 2013-08-07 | 大连海事大学 | Microalgae activity detection method and device in ship ballast water |
| CN103335945A (en) * | 2009-07-07 | 2013-10-02 | 索尼公司 | Microfluidic device |
| CN103364382A (en) * | 2013-07-12 | 2013-10-23 | 大连海事大学 | Ship domestic sewage detection device |
| CN105136763A (en) * | 2015-09-10 | 2015-12-09 | 大连海事大学 | Single microalgae cell activity dynamic monitoring novel method and device based on gas-liquid interface single cell capture and chlorophyll fluorescence characterization |
| CN105203468A (en) * | 2015-10-19 | 2015-12-30 | 中国科学院合肥物质科学研究院 | Micro-fluidic chip system for improving radiation flux of single ion beam |
| CN106190829A (en) * | 2016-07-26 | 2016-12-07 | 西安交通大学 | A kind of for cell high accuracy arrangement and the microflow controlled biochip of detection |
| CN106483059A (en) * | 2016-10-13 | 2017-03-08 | 中国人民解放军第三军医大学第附属医院 | Terahertz fluidic cell sensor and its detection method for single or a small amount of living cells markless detection |
| CN107991278A (en) * | 2017-12-28 | 2018-05-04 | 湖南华南光电科技股份有限公司 | Heating sampling structure based on fluorescent quenching technology trace explosive detector |
| CN112229780A (en) * | 2020-09-07 | 2021-01-15 | 桂林电子科技大学 | Improved flow cytometer based on optical fiber integrated microfluidic chip |
| CN112697679A (en) * | 2020-12-04 | 2021-04-23 | 杭州娃哈哈精密机械有限公司 | Device for rapidly detecting number of bacteria in beverage |
| CN113049478A (en) * | 2021-04-25 | 2021-06-29 | 中国计量科学研究院 | Protein aggregate analysis and detection device based on micro-fluidic chip and working method |
| CN113866075A (en) * | 2021-08-13 | 2021-12-31 | 河海大学 | Method for rapidly determining volume of microcystis pseudo-vacuoles by using flow cytometer |
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2005
- 2005-06-19 CN CN 200520084372 patent/CN2821565Y/en not_active Expired - Fee Related
Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103335945A (en) * | 2009-07-07 | 2013-10-02 | 索尼公司 | Microfluidic device |
| CN102507528A (en) * | 2011-12-02 | 2012-06-20 | 徐州雷奥医疗设备有限公司 | Micro-fluid detection system based on self-focusing lens |
| CN103234949A (en) * | 2013-01-30 | 2013-08-07 | 大连海事大学 | Microalgae activity detection method and device in ship ballast water |
| CN103364382A (en) * | 2013-07-12 | 2013-10-23 | 大连海事大学 | Ship domestic sewage detection device |
| CN105136763B (en) * | 2015-09-10 | 2017-10-13 | 大连海事大学 | The active dynamic monitoring new method of single microalgae cell and device characterized based on the unicellular capture of gas-liquid interface and chlorophyll fluorescence |
| CN105136763A (en) * | 2015-09-10 | 2015-12-09 | 大连海事大学 | Single microalgae cell activity dynamic monitoring novel method and device based on gas-liquid interface single cell capture and chlorophyll fluorescence characterization |
| CN105203468A (en) * | 2015-10-19 | 2015-12-30 | 中国科学院合肥物质科学研究院 | Micro-fluidic chip system for improving radiation flux of single ion beam |
| CN105203468B (en) * | 2015-10-19 | 2018-03-23 | 中国科学院合肥物质科学研究院 | A kind of micro-fluidic chip system for being used to improve Single ion microbeam fluence |
| CN106190829B (en) * | 2016-07-26 | 2018-07-03 | 西安交通大学 | A kind of microflow controlled biochip for arranging and detecting for cell high-precision |
| CN106190829A (en) * | 2016-07-26 | 2016-12-07 | 西安交通大学 | A kind of for cell high accuracy arrangement and the microflow controlled biochip of detection |
| CN106483059A (en) * | 2016-10-13 | 2017-03-08 | 中国人民解放军第三军医大学第附属医院 | Terahertz fluidic cell sensor and its detection method for single or a small amount of living cells markless detection |
| CN107991278A (en) * | 2017-12-28 | 2018-05-04 | 湖南华南光电科技股份有限公司 | Heating sampling structure based on fluorescent quenching technology trace explosive detector |
| CN107991278B (en) * | 2017-12-28 | 2024-04-26 | 湖南华南光电科技股份有限公司 | Heating sample injection structure of trace explosive detector based on fluorescence quenching technology |
| CN112229780A (en) * | 2020-09-07 | 2021-01-15 | 桂林电子科技大学 | Improved flow cytometer based on optical fiber integrated microfluidic chip |
| CN112697679A (en) * | 2020-12-04 | 2021-04-23 | 杭州娃哈哈精密机械有限公司 | Device for rapidly detecting number of bacteria in beverage |
| CN113049478A (en) * | 2021-04-25 | 2021-06-29 | 中国计量科学研究院 | Protein aggregate analysis and detection device based on micro-fluidic chip and working method |
| CN113049478B (en) * | 2021-04-25 | 2022-05-17 | 中国计量科学研究院 | Protein aggregate analysis and detection device based on micro-fluidic chip and working method |
| CN113866075A (en) * | 2021-08-13 | 2021-12-31 | 河海大学 | Method for rapidly determining volume of microcystis pseudo-vacuoles by using flow cytometer |
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