CN103614283A - Microfluidic chip for three-dimensional enrichment of cells - Google Patents
Microfluidic chip for three-dimensional enrichment of cells Download PDFInfo
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- CN103614283A CN103614283A CN201310597726.3A CN201310597726A CN103614283A CN 103614283 A CN103614283 A CN 103614283A CN 201310597726 A CN201310597726 A CN 201310597726A CN 103614283 A CN103614283 A CN 103614283A
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- 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
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Abstract
The invention relates to a microfluidic chip for the three-dimensional enrichment of cells. The microfluidic chip consists of a cover glass layer and slide glass which is adhered below the cover glass layer, wherein a microchannel is arranged in the cover glass layer and comprises a bent channel, a symmetrical-apex-angle-structured channel, an optical detection zone and a liquid discharge zone, wherein the bent channel is used for enriching cells in the vertical direction, the symmetrical-apex-angle-structured channel is communicated with the bent channel and is used for enriching the cells in the horizontal direction, the optical detection zone is communicated with the symmetrical-apex-angle-structured channel, and the liquid discharge zone is communicated with the optical detection zone. The microchannel further comprises a cell sample liquid adding zone and a buffer solution adding zone which are communicated with an inlet of the bent channel, wherein the cell sample liquid adding zone is communicated with the internal wall of the bent channel, and the buffer solution adding zone is communicated with the external wall of the bent channel; the symmetrical-apex-angle-structured channel consists of a plurality of expansion sections and symmetrical apex-angle structures, which are distributed alternately; an inlet of the buffer solution adding zone, an inlet of the cell sample liquid adding zone and the liquid discharge zone are through holes formed in the cover glass layer, the microchannel is a blind channel formed in the cover glass layer, and the bottom of the microchannel is communicated with the surface of the slide glass. The microfluidic chip has the characteristics of simple structure, low cost, simplicity and easiness in operation, small size, convenience in popularization, and the like.
Description
Technical field
The present invention relates to a kind of micro-fluidic chip, be specifically related to a kind of micro-fluidic chip for cell three-dimensional enrichment.
Background technology
In biomedical research and clinical diagnosis, flow cytometry instrument is widely used, and has brought into play very important effect.In recent years, it becomes a kind of main method of clinical diagnosis aspect, for example, and CD
4 +t subset lymphocyte count instrument is widely used in detection and the diagnosis of the HIV virus in blood.When HIV (human immunodeficiency virus) (HIV) enters after human body, can attack specifically CD
4 +t lymphocyte also carries out self-replication, causes CD in blood
4 +the reduction of T lymphocyte function and the minimizing of quantity.Lymphocyte is as the important composition of immune response function, after being destroyed, will cause the continuous weakening of host immune function by HIV virus, thus HIV virus lays dormant after 2～8 years because opportunistic infection are fallen ill.CD
4 +t subset lymphocyte count instrument passes through the CD in blood
4 +t lymphocyte detects, thereby for HIV progression of disease situation and HAART clinical efficacy carry out Efficient Evaluation, for the Clinics and Practices of HIV disease provides important references.In addition, flow cytometry instrument is also used to conventional blood test.In the major function of blood if matter transportation and body defence etc. is all that hemocyte in blood completes.The transportation function of blood mainly completes by red corpuscle, as all relying on blood transportation, the nutritive substance of the oxygen sucking from lung and digestive tube absorption could arrive each tissue of whole body, the carbonic acid gas that tissue metabolism produces simultaneously and other refuses also rely on blood transportation and locate excretion to lung, kidney etc., thereby keep the carrying out of health eubolism.Blood shows as immunity and hemostasis etc. to the defense function of body.As the white corpuscle in blood can kill pathogenic agent and self abnormal cells that enters body, thus performance body immunity; Thrombocyte plays an important role in hemostasis, and in normal physiological situation, thrombocyte has metastable quantity.When the platelet content in blood changes, the hemostasis of human body and coagulation function will occur extremely, entail dangers to patient's life security when serious.By detecting hematoblastic content in blood, correct evaluator body hemostasis and coagulation function are had great significance.Detected result can be used as the reference index whether patient needs to carry out platelet transfusion simultaneously.Thereby use flow cytometry instrument to detect hemocyte in blood etc., just can reflect the healthy state of human body, thereby provide important reference frame for health care and disease prevention.In flow cytometry instrument, cell to be detected need to be dimensionally enriched in the center of surveyed area, then through laser detection region, and then can obtain many information such as type, quantity, size of cell to be checked.An important cell enrichment in flow cytometry instrument is partly coaxial annular runner, effect by circumferential shearing fluid by cell three-dimensional to be measured be enriched in channel center and be closely aligned into a branch of, thereby avoided undetected in laser detection district of cell to be measured.Yet, in traditional flow cytometry instrument, the three-dimensional enrichment of cell has following shortcoming: 1) need expensive supplementary instrument, thereby cause flow cytometry instrument holistic cost to raise, greatly limited its application in the fields such as biomedicine, clinical diagnosis; 2) coaxial annular runner and supplementary instrument volume are large, cause traditional blood-counter system to be not easy to carry, thereby can not be conveniently used in the aspects such as field first aid, domestic medicine and individual health care, and commercial marketing is brought to difficulty; 3) cell three-dimensional enrichment complex structure, installs loaded down with trivial detailsly, and maintenance expense is higher, has increased the cost that the later stage used.
Summary of the invention
For solving above-mentioned problems of the prior art, the object of the present invention is to provide a kind of micro-fluidic chip for cell three-dimensional enrichment, it can be as the preprocessing means of blood testing, replace baroque cell enrichment device in the mobile blood-counter system of tradition, and can combine with optical detection means, blood sample is accurately detected, have that cost is low, volume is little, simple in structure, the features such as processing ease, for the fields such as public medical health and biomedical research facilitate.
In order to achieve the above object, the present invention adopts following technical scheme:
For a micro-fluidic chip for cell three-dimensional enrichment, described micro-fluidic chip is comprised of cover plate layer 8 and the slide glass 7 being bonded under cover plate layer 8; In described cover plate layer 8, be provided with microchannel, described microchannel comprise make optical detection zone 5 that the symmetrical horn structure passage 4 that makes cell enrichment in the horizontal direction that the bending channel 3 of cell in the vertical direction enrichment and bending channel 3 be communicated with and symmetrical horn structure passage 4 be communicated with and and the drainage region 6 that is communicated with, optical detection zone 5, also comprise that the cell sample liquid being communicated with bending channel 3 entrances adds liquid zone 2 and damping fluid adds liquid zone 1, described cell sample liquid adds the inwall that liquid zone 2 is communicated in bending channel 3, and described damping fluid adds the outer wall that liquid zone 1 is communicated in bending channel 3; Described symmetrical horn structure passage 4 is comprised of a plurality of expansion segments that distribute alternately and symmetrical horn structure; Described damping fluid adds entrance and drainage region 6 through hole for offering on cover plate layer 8 that liquid zone 1 and cell sample liquid add liquid zone 2, the sidewalk for visually impaired people of described microchannel for offering on cover plate layer 8, and bottom and slide glass 7 surfaces communicate.
Described bending channel 3 is 1/4 circular arc, and its width is 100 μ m, and mean radius is 250 μ m.
Described symmetrical horn structure passage 4 is comprised of 30 groups of symmetrical horn structure and expansion segment.
The wedge angle of described symmetrical horn structure is 45 degree, and symmetrical wedge angle entrance width is 40 μ m, and the width of described expansion segment is 300 μ m, and length is 200 μ m.
Described cover plate layer 8 and slide glass 7 are bonded together by the irreversible processing of plasma.
Described damping fluid adds entrance and the drainage region 6 that liquid zone 1 and cell sample liquid adds liquid zone 2 and is cylindrical hole.
Described damping fluid adds liquid zone 1 and cell sample liquid adds the entrance of liquid zone 2 and the diameter of drainage region 6 is 10mm.
The described microchannel everywhere degree of depth is 78 μ m.
The material of described cover plate layer 8 is polymetylmethacrylate or polydimethylsiloxane.
The material of described slide glass 7 is glass or silicon.
Compared to the prior art, tool has the following advantages in the present invention:
1) volume is little, is easy to carry, and can be conveniently used in the aspects such as field first aid, domestic medicine and individual health care.
2) cost is low, cheap, compares with other traditional and active microfluid cell three-dimensional beneficiation technologies, and the present invention is without expensive utility appliance, thereby processing and making process is simple and convenient, and commercial marketing is offered convenience.
3) physical construction is simple, and processing ease is compared with other the three-dimensional beneficiation technologies of passive microfluid, carry out accurate flow control, thereby processing ease is simple without the damping fluid to more, and operator are without professional training.
4) extensibility is high, and owing to having adopted the form of micro-fluidic chip, the present invention can be integrated with sample post-processing technology, promotes the integrated and automatization of analytical system.As combined with optical detection means, just can detect accurately target compounds such as cells, thus in fields such as genetic expression detection, genescreen, drug screening, medical diagnosis on disease treatment, environment measuring and improvement, judicial expertises, play an important role.
Accompanying drawing explanation
Fig. 1 is the vertical view of micro-fluidic chip of the present invention.
Fig. 2 be Fig. 1 along A-A to sectional view.
Fig. 3 is the microchannel schematic three dimensional views in micro-fluidic chip of the present invention.
Fig. 4 is that the present invention is in conjunction with the integrated schematic diagram of optical detection means.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is further detailed explanation.
As shown in Figure 1 to Figure 3, a kind of micro-fluidic chip for cell three-dimensional enrichment of the present invention, this micro-fluidic chip is processed by standard soft lithography, cover plate layer 8 and the slide glass 7 being bonded under cover plate layer 8, consists of; In cover plate layer 8, be provided with microchannel, microchannel comprise make optical detection zone 5 that the symmetrical horn structure passage 4 that makes cell enrichment in the horizontal direction that the bending channel 3 of cell in the vertical direction enrichment and bending channel 3 be communicated with and symmetrical horn structure passage 4 be communicated with and and the drainage region 6 that is communicated with, optical detection zone 5, also comprise that the cell sample liquid being communicated with bending channel 3 entrances adds liquid zone 2 and damping fluid adds liquid zone 1, cell sample liquid adds the inwall that liquid zone 2 is communicated in bending channel 3, and damping fluid adds the outer wall that liquid zone 1 is communicated in bending channel 3; Symmetrical horn structure passage 4 is comprised of a plurality of expansion segments that distribute alternately and symmetrical horn structure; Damping fluid adds entrance and drainage region 6 through hole for offering on cover plate layer 8 that liquid zone 1 and cell sample liquid add liquid zone 2, the sidewalk for visually impaired people of microchannel for offering on cover plate layer 8, and bottom and slide glass 7 surfaces communicate.
As the preferred embodiment of the present invention, described bending channel 3 is 1/4 circular arc.Further, described bending channel 3 width are 100 μ m, and mean radius is 250 μ m.
As the preferred embodiment of the present invention, described symmetrical horn structure passage 4 is comprised of 30 groups of symmetrical horn structure and expansion segment.Further, the wedge angle of described symmetrical horn structure is 45 degree, and symmetrical wedge angle entrance width is 40 μ m, and the width of described expansion segment is 300 μ m, and length is 200 μ m.
As the preferred embodiment of the present invention, described cover plate layer 8 and slide glass 7 are bonded together by the irreversible processing of plasma.
As the preferred embodiment of the present invention, described damping fluid adds entrance and the drainage region 6 that liquid zone 1 and cell sample liquid adds liquid zone 2 and is cylindrical hole.Further, described damping fluid adds liquid zone 1 and cell sample liquid adds the entrance of liquid zone 2 and the diameter of drainage region 6 is 10mm.
As the preferred embodiment of the present invention, the material of described cover plate layer 8 is polymetylmethacrylate or polydimethylsiloxane.
As the preferred embodiment of the present invention, the material of described slide glass 7 is glass or silicon.
Principle of work of the present invention is: the sample liquid that contains cell adds by cell sample liquid the inner side that liquid zone 2 enters bending channel 3 with certain flow, and damping fluid adds by damping fluid the outside that liquid zone 1 enters bending channel 3 with certain flow.When cell sample liquid and damping fluid are flowed through bending channel 3, inertia effect due to fluid, fluid particle can be to outside wall surface one lateral movement of bending channel 3, and on microchannel, lower wall surface has limitations affect near fluid it simultaneously, thereby forms certain pressure gradient on the cross section of bending channel 3.Under the effect of pressure gradient, the fluid of channel center is to outside wall surface one lateral movement of bending channel 3, near fluid on passage lower wall surface is as inner-wall surface one lateral movement to bending channel 3, thereby on the cross section at bending channel 3 perpendicular to flow direction, form the whirlpool, Dean of two symmetrical reverse, under the strong effect of dragging of Dean vortex strength, the cell sample liquid that is positioned at bending channel 3 inner sides just by from inner-wall surface one side via channel center to outside wall surface one side stretching, outlet at bending channel 3, cell sample liquid is just distributed in the central position of channel cross-section in the vertical direction, realize the enrichment of cell in the vertical direction.Then, in the vertical direction, by the cell sample liquid of enrichment, flows through and has symmetrical horn structure passage 4, and certain displacement occurs in the centrifugal inertial force ，Xiang channel center that the direction that is subject to being caused by symmetrical horn structure is pointed to channel center by both sides.Due to storage effect, after a series of symmetrical horn structure, cell sample liquid will fully move to channel center in the horizontal direction, realizes cell enrichment in the horizontal direction.In conjunction with bending micro 3 and symmetrical horn structure passage 4, finally realize the three-dimensional enrichment of cell.By the cell of three-dimensional enrichment, by symmetrical horn structure passage 4 outlets, entered optical detection zone 5, in optical detection zone, the 5 accurate detections that realize cell, are finally discharged by leakage fluid dram 6.
As shown in Figure 4, below with the CD in the detection of HIV virus
4 +the three-dimensional enrichment of T lymphocyte is that example illustrates the specific embodiment of the present invention:
A. in patient body, extract blood sample, and carry out necessary pre-treatment;
B. processed mistake is contained to CD
4 +the lymphocytic blood sample of T adds liquid zone 2 entrances with certain uninterrupted by cell sample liquid and injects micro-fluidic chip of the present invention under the driving of the driving arrangements such as syringe pump, and simultaneous buffering liquid injects damping fluid with certain flow and adds liquid zone 1 entrance;
C. CD will be contained in the whirlpool, Dean in bending channel 3
4 +the lymphocytic blood sample of T is to bending channel outside wall surface one side stretching, thereby realizes the CD in blood
4 +the enrichment of T lymphocyte in the vertical direction;
D. on vertical direction by the CD after enrichment
4 +t lymphocyte enters symmetrical horn structure passage 4, and the direction that is subject to wedge angle initiation is pointed to the centrifugal inertial force at center, microchannel, thereby realizes the CD in blood
4 +the enrichment in the horizontal direction of T lymphocyte; At the outlet of symmetrical horn structure passage 4, CD
4 +t lymphocyte is dimensionally enriched in the central position of passage;
E. by the CD of three-dimensional enrichment
4 +t lymphocyte then enters optical detection zone 5, in conjunction with certain optical detection means, completes CD
4 +the lymphocytic detection of T, also can be integrated other the detection chip CD to three-dimensional enrichment
4 +t lymphocyte detects;
F. the blood sample after being detected is discharged by leakage fluid dram 6, carries out necessary aftertreatment, to the HIV inactivation of virus that may contain in blood sample, guarantees safety.
Claims (10)
1. for a micro-fluidic chip for cell three-dimensional enrichment, it is characterized in that: by cover plate layer (8) and the slide glass (7) that is bonded under cover plate layer (8), formed, in described cover plate layer (8), be provided with microchannel, described microchannel comprises the bending channel (3) that makes the enrichment of cell in the vertical direction, and the symmetrical horn structure passage (4) that makes cell enrichment in the horizontal direction of bending channel (3) connection, optical detection zone (5) with symmetrical horn structure passage (4) connection, and and optical detection zone (5) drainage region (6) of being communicated with, also comprise and cell sample liquid that bending channel (3) entrance is communicated with adds liquid zone (2) and damping fluid adds liquid zone (1), described cell sample liquid adds the inwall that liquid zone (2) is communicated in bending channel (3), described damping fluid adds the outer wall that liquid zone (1) is communicated in bending channel (3), described symmetrical horn structure passage (4) is comprised of a plurality of expansion segments that distribute alternately and symmetrical horn structure, described damping fluid adds entrance and drainage region (6) through hole for offering on cover plate layer (8) that liquid zone (1) and cell sample liquid add liquid zone (2), the sidewalk for visually impaired people of described microchannel for offering on cover plate layer (8), and bottom and slide glass (7) surface communicates.
2. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 1, is characterized in that: described bending channel (3) is 1/4 circular arc, and its width is 100 μ m, and mean radius is 250 μ m.
3. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 1, is characterized in that: described symmetrical horn structure passage (4) is comprised of 30 groups of symmetrical horn structure and expansion segment.
4. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 3, is characterized in that: the wedge angle of described symmetrical horn structure is 45 degree, and symmetrical wedge angle entrance width is 40 μ m, and the width of described expansion segment is 300 μ m, and length is 200 μ m.
5. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 1, is characterized in that: described cover plate layer (8) and slide glass (7) are bonded together by the irreversible processing of plasma.
6. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 1, is characterized in that: described damping fluid adds entrance and drainage region (6) that liquid zone (1) and cell sample liquid adds liquid zone (2) and is cylindrical hole.
7. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 6, is characterized in that: described damping fluid adds liquid zone (1) and cell sample liquid adds the entrance of liquid zone (2) and the diameter of drainage region (6) is 10mm.
8. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 1, is characterized in that: the described microchannel everywhere degree of depth is 78 μ m.
9. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 1, is characterized in that: the material of described cover plate layer (8) is polymetylmethacrylate or polydimethylsiloxane.
10. a kind of micro-fluidic chip for cell three-dimensional enrichment according to claim 1, is characterized in that: the material of described slide glass (7) is glass or silicon.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| CN106215985A (en) * | 2016-07-26 | 2016-12-14 | 西安交通大学 | A kind of micro-fluidic chip quickly mixing for fluid and detecting |
| CN107376750A (en) * | 2017-08-03 | 2017-11-24 | 青岛科技大学 | A kind of micro-fluid chip that can be achieved efficiently to mix |
| CN110871137A (en) * | 2019-11-27 | 2020-03-10 | 中国矿业大学 | Small-particle-size fly ash particle sorting spiral runner microfluidic device and method |
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| CN101765762A (en) * | 2007-04-16 | 2010-06-30 | 通用医疗公司以马萨诸塞州通用医疗公司名义经营 | Systems and methods for particle focusing in microchannels |
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| CN1635146A (en) * | 2004-09-23 | 2005-07-06 | 湖南大学 | One-dimensional biological chip and application in gene, protein expression analysis |
| CN101765762A (en) * | 2007-04-16 | 2010-06-30 | 通用医疗公司以马萨诸塞州通用医疗公司名义经营 | Systems and methods for particle focusing in microchannels |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
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| CN110871137A (en) * | 2019-11-27 | 2020-03-10 | 中国矿业大学 | Small-particle-size fly ash particle sorting spiral runner microfluidic device and method |
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