CN110339876A - A kind of tumour cell screening micro fluidic device and screening method based on drop deposit - Google Patents
A kind of tumour cell screening micro fluidic device and screening method based on drop deposit Download PDFInfo
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- CN110339876A CN110339876A CN201910564355.6A CN201910564355A CN110339876A CN 110339876 A CN110339876 A CN 110339876A CN 201910564355 A CN201910564355 A CN 201910564355A CN 110339876 A CN110339876 A CN 110339876A
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- 210000004881 tumor cell Anatomy 0.000 title claims abstract description 49
- 238000012216 screening Methods 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 15
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 34
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 19
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims abstract description 19
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims abstract description 19
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 19
- 239000011521 glass Substances 0.000 claims abstract description 6
- 238000011897 real-time detection Methods 0.000 claims abstract description 5
- 239000000758 substrate Substances 0.000 claims abstract description 5
- 238000012360 testing method Methods 0.000 claims abstract description 5
- 210000004027 cell Anatomy 0.000 claims description 80
- WSNMPAVSZJSIMT-UHFFFAOYSA-N COc1c(C)c2COC(=O)c2c(O)c1CC(O)C1(C)CCC(=O)O1 Chemical compound COc1c(C)c2COC(=O)c2c(O)c1CC(O)C1(C)CCC(=O)O1 WSNMPAVSZJSIMT-UHFFFAOYSA-N 0.000 claims description 18
- 238000005452 bending Methods 0.000 claims description 10
- 210000004369 blood Anatomy 0.000 claims description 9
- 239000008280 blood Substances 0.000 claims description 9
- HHBBIOLEJRWIGU-UHFFFAOYSA-N 4-ethoxy-1,1,1,2,2,3,3,4,5,6,6,6-dodecafluoro-5-(trifluoromethyl)hexane Chemical compound CCOC(F)(C(F)(C(F)(F)F)C(F)(F)F)C(F)(F)C(F)(F)C(F)(F)F HHBBIOLEJRWIGU-UHFFFAOYSA-N 0.000 claims description 4
- 238000001125 extrusion Methods 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 238000003860 storage Methods 0.000 claims description 3
- 238000000151 deposition Methods 0.000 claims description 2
- 238000001727 in vivo Methods 0.000 claims 1
- 239000007787 solid Substances 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 17
- 239000007788 liquid Substances 0.000 abstract description 7
- 239000012530 fluid Substances 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000005070 sampling Methods 0.000 abstract description 2
- 238000004458 analytical method Methods 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 201000010099 disease Diseases 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 239000000571 coke Substances 0.000 description 2
- 238000007334 copolymerization reaction Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 241000406668 Loxodonta cyclotis Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
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- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000012631 diagnostic technique Methods 0.000 description 1
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- 238000007877 drug screening Methods 0.000 description 1
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Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
- B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/12—Specific details about manufacturing devices
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- G01N2015/1022—
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- G01N2015/1027—
Abstract
The present invention relates to a kind of tumour cell screening micro fluidic devices and screening method based on drop deposit, belong to microfluidic art.The present invention includes glass substrate and the PDMS chip that is fixed thereon, wherein PDMS material constitutes entire chip, microchannel structure is made by micrometer-nanometer processing technology, a drop, which is generated, by cross-shaped configuration wraps up an erythrocyte or tumour cell, then passive valve is deposited by drop, due to the size and softness difference of erythrocyte and tumour cell, the drop of the drop and package tumour cell that wrap up erythrocyte is different by the number of drop register, the number passed through by laser detection drop, to detect whether tumour cell, realize the highly sensitive real-time detection of few samples amount.The present invention, which realizes, is integrated in sampling fluids, driving, manipulation, transport and testing and analyzing on micro-fluidic chip, realizes that liquid flows controllable, micro and trace materials transport and efficient detection.
Description
Technical field
The present invention relates to a kind of tumour cell screening micro fluidic devices and screening method based on drop deposit, belong to miniflow
Control technical field.
Background technique
Micro-fluidic chip and it is widely used in the fields such as medical diagnosis on disease, drug screening, environment measuring, food safety.It is micro-
One of bare bones of fluidic chip are exactly operated and are controlled to the fluid under minute yardstick, and pair as operation and control
The Fluid Volume of elephant is again extremely small, and many forms of expression and constant substance of substance are different.Tumour disease is to threaten the mankind raw
Most dangerous one of the disease of life health, the basic reason generated are that control growth and proliferation of cell mechanism is not normal, i.e., wither with cell
The exception died is related, caused by the traditional medical detecting method of tumour cell is lower to its detection efficiency.Therefore, how quickly, it is quasi-
True is the pressing problem of modern biology research to tumour cell progress quantitative detection and sorting.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of tumour cell screening micro fluidic devices based on drop deposit
And screening method, it can be realized micro and constant substance transport and efficient detection.
The technical solution adopted by the present invention: a kind of tumour cell screening micro fluidic device based on drop deposit, including glass
Glass substrate 1 and PDMS chip 2, the controller being fixed thereon, contain on PDMS chip 2 continuous phase entrance 4, focusing structure 5,
Continuous phase outlet 7, dispersed phase entrance 6, register channel, outlet 3, focusing structure 5 be crisscrossing channels, the two of focusing structure 5
A arrival end is connect with continuous phase entrance 4, dispersed phase entrance 6 respectively by microchannel, and two outlet ends are distinguished by microchannel
Connect with continuous phase outlet 7 and the entrance in register channel, the dispersed phase that enters from dispersed phase entrance 6 and from continuous phase entrance 4
The continuous phase of entrance generates package cell drop 8 by focusing structure 5 altogether, and package cell drop 8 passes through focusing structure 5 and deposit
Microchannel between the entrance in device channel enters register channel, register channel be upward continuous bending shape and its in from forward direction
It is successively arranged the register that several structures are identical, length gradually shortens afterwards, register is connect with controller, two neighboring deposit
Device is staggeredly located at the bending part in register channel up and down, wraps up cell drop 8 with fixed frequency and passes sequentially through each register,
The number of the detectable package cell drop 8 by it of each register, the outlet in register channel passes through microchannel and outlet 3
Connection.
Preferably, the outlet 3, continuous phase entrance 4, dispersed phase entrance 6, continuous phase export 7 proper alignments in PDMS
The right side of PDMS chip 2 is arranged in the left side of chip 2, register channel.
Specifically, be successively arranged from front to back in the register channel drop register I 9, drop register II 10,
Drop register III 11, drop register IV 12, drop register V 13, drop register VI 14, adjacent drops register it
Between by microchannel connect, drop register V 13 include deposit chamber, laser detector 13-5, laser detector 13-5 and control
Device connection, deposit chamber export 13-4 including deposit chamber entrance 13-2, cavity, deposit chamber, and deposit chamber entrance 13-2 is micro- logical with front end
Road connection, deposit chamber outlet 13-4 and bending part vertical microchannel connect, laser detector 13-5 be mounted on deposit chamber just on
In the microchannel of side, the width for depositing chamber is greater than the width of preceding latter linked microchannel, deposits and is equipped in the cavity of chamber and cavity
I 13-1 of rectangle supplementary structure, II 13-3 of rectangle supplementary structure that inner wall is spaced apart, rectangle supplementary structure II 13-3
In length of the end and its length of I 13-1 of rectangle supplementary structure greater than I 13-1 of rectangle supplementary structure, rectangle auxiliary knot
I 13-1 of structure includes several groups rectangular slab symmetrical above and below and horizontal positioned, adjacent two rectangles auxiliary knot in same horizontal line
I 13-1 of structure is arranged at equal intervals, and the width between opposite two rectangle supplementary structures, I 13-1 is equal to the width of front end microchannel, long
Rectangular II 13-3 of supplementary structure includes orthogonal one horizontal positioned rectangular slab and the rectangular slab that is disposed vertically, is hung down
II 13-3 of rectangle supplementary structure directly placed is located at the top of horizontal positioned II 13-3 of rectangle supplementary structure and vertically puts
The end of I 13-1 of rectangle supplementary structure of II 13-3 of rectangle supplementary structure and top that set form notch, into deposit chamber
Interior package cell drop 8 enters the microchannel of rear end after notch extrusion from deposit chamber outlet 13-4.
Specifically, a length of 120mm of the PDMS chip 2, width 60mm, a height of 10mm；The width of all microchannels is
0.020mm, a height of 0.010mm.
Specifically, the drop register I 9, drop register II 10, drop register III 11, drop register IV 12,
In drop register V 13, drop register VI 14 deposit chamber cavity length in the horizontal direction be respectively 0.180mm,
0.160mm,0.140mm,0.120mm,0.100mm,0.080mm；The width of each register deposit chamber cavity is 0.040mm, high
It is 0.010mm；The a length of 0.010mm of I 13-1 of rectangle supplementary structure, width 0.003mm, quantity is 8；Rectangle auxiliary knot
II 13-3 of structure a length of 0.020mm, width 0.003mm are divided into same horizontal line between adjacent two rectangles supplementary structure, I 13-1
0.005mm；Width between I 13-1 of rectangle supplementary structure, II 13-3 of rectangle supplementary structure and corresponding register cavity inner wall
Degree is 0.005mm；The end of I 13-1 of rectangle supplementary structure of II 13-3 of rectangle supplementary structure and top that are disposed vertically
The width for forming notch is 0.005mm.
A kind of screening method of the tumour cell screening micro fluidic device based on drop deposit, specific steps are such as
Under:
Step1: injecting fluorinated oil HFE-7500 to continuous phase entrance 4, and the injection of dispersed phase entrance 6 has the blood of tumour cell,
By controlling the entrance velocity of dispersed phase entrance 6 and continuous phase entrance 4 respectively, when normal erythrocyte enters cross copolymerization coke
After structure 5, the package cell drop 8 with single blood erythrocyte by mocro of 0.020mm diameter can be stably generated, when bigger than erythrocyte
Or after small tumour cell enters cross focusing structure 5 altogether, the package cell liquid than having single blood erythrocyte by mocro can be generated
Drip 8 big or small package cell drops 8, package cell drop 8 pass sequentially through drop register I 9, drop register II 10,
Drop register III 11, drop register IV 12, drop register V 13, drop register VI 14, each drop register energy
The quantity for accommodating package cell drop 8 is the packet beyond receiving number that is fixed, finally entering in each drop register
The drop register can be squeezed out a drop of foremost by wrapping up in cell drop 8, the laser detector in each drop register
The number for detecting the package cell drop 8 by respective drop register, and will test result and sends controller to, from last
The package cell drop 8 of one drop register outflow flow to outside from outlet 3；
Step2: due to size and the softness difference of erythrocyte and tumour cell, each drop register is squeezing package
Required number is different when the package cell drop 8 of different cells, is stored in controller all normal in the set time
By the quantity of the package cell drop 8 of each drop register when erythrocyte passes through, controller is according to each drop register
In the real-time detection structure of laser detector judge whether contain tumour cell in blood, in the same time, when by each
Pass through the packet of each drop register when the quantity of the package cell drop 8 of drop register and all normal erythrocytes pass through
The quantity for wrapping up in cell drop 8 is different, then it is assumed that there are tumour cells.
Drop register I 9, drop register II 10, drop register III 11, drop register IV 12, drop register
V 13, the quantity that drop register VI 14 can accommodate the package cell drop 8 with single blood erythrocyte by mocro is respectively 7,6,5
A, 4,3,2.
The beneficial effects of the present invention are:
1, present apparatus structure be simple and convenient to operate, high sensitivity.
2, dispersed phase uses fluorinated oil HFE-7500, and fluorinated oil has good bio-compatibility, to what is wrapped up in drop
The biologic grains such as cell, virus do not have toxic effect, guarantee that biological respinse can be normally carried out in drop.
3, the present invention is led to using mechanical analysis instead of blood biochemical check analysis used in traditional medical diagnostic techniques
The detection that the drop of multiple drop registers passes through is crossed, the highly sensitive real-time detection of few samples amount may be implemented.
4, analysis time of the invention is short, sample consumption is few, highly sensitive and cost is relatively low, can quickly detect whether
There is tumour cell.
Detailed description of the invention
Fig. 1 is overall structure diagram of the invention；
Fig. 2 is the enlarged structure schematic diagram of drop register V in Fig. 1；
Fig. 3 is the quantity that each drop register stores A tumour cell, normal erythrocyte, B tumour cell in the embodiment of the present invention
Distribution map.
Each label in figure are as follows: glass substrate -1, PDMS chip -2, outlet -3, continuous phase entrance -4, altogether focusing structure -5,
Dispersed phase entrance -6, continuous phase outlet -7, package cell drop -8, drop register I -9, drop register II -10, drop are posted
Storage III -11, drop register IV -12, drop register V -13, drop register VI -14, I -13- of rectangle supplementary structure
1, chamber entrance -13-2, II -13-3 of rectangle supplementary structure, deposit chamber outlet -13-4, laser detector -13-5 are deposited.
Specific embodiment
In the following with reference to the drawings and specific embodiments, the present invention is described further.
Embodiment 1: a kind of tumour cell screening micro fluidic device based on drop deposit, including glass substrate 1 and fixation
PDMS chip 2, controller on it, contain on PDMS chip 2 continuous phase entrance 4, focusing structure 5, continuous phase outlet 7,
Dispersed phase entrance 6, register channel, outlet 3, focusing structure 5 are crisscrossing channels, and the two entrances end of focusing structure 5 passes through
Microchannel is connect with continuous phase entrance 4, dispersed phase entrance 6 respectively, and two outlet ends export 7 with continuous phase respectively by microchannel
And the entrance connection in register channel, the dispersed phase entered from dispersed phase entrance 6 are passed through with the continuous phase entered from continuous phase entrance 4
Cross focusing structure 5 altogether and generate package cell drop 8, package cell drop 8 by the entrance in focusing structure 5 and register channel it
Between microchannel enter register channel, register channel is upward continuous bending shape (as shown in Figure 1, for upward continuous square wave
Shape bending) and the register that several structures are identical, length gradually shortens, register and control are successively arranged in it from front to back
Device connection, two neighboring register are staggeredly located at the bending part in register channel up and down, wrap up cell drop 8 with fixed frequency
Each register is passed sequentially through, the number of the detectable package cell drop 8 by it of each register, register channel goes out
Mouth is connect by microchannel with outlet 3.
Further, the outlet 3, continuous phase entrance 4, dispersed phase entrance 6, continuous phase export 7 proper alignments and exist
The right side of PDMS chip 2 is arranged in the left side of PDMS chip 2, register channel.Export 3, continuous phase entrance 4, dispersed phase entrance
6, it is that insertion capillary is passed through liquid for convenience on the left of chip that continuous phase, which exports 7 proper alignments,.Due to each register quilt
The package cell drop 8 extruded exports upwards, so each register to be alternately arranged in series in the right side of PDMS chip 2 upwards
Side.
Further, it is successively arranged drop register I 9, drop register II from front to back in the register channel
10, drop register III 11, drop register IV 12, drop register V 13, drop register VI 14, adjacent drops register
Between connected by microchannel, as shown in Fig. 2, since each drop register architecture is identical, only by taking drop register V 13 as an example,
The structure of a drop register is described in detail: drop register V 13 includes deposit chamber, laser detector 13-5, laser
Detector 13-5 is connect with controller, and deposit chamber includes deposit chamber entrance 13-2, cavity, deposit chamber outlet 13-4, and deposit chamber enters
Mouth 13-2 is connect with front end microchannel, and the vertical microchannel of deposit chamber outlet 13-4 and bending part connects, laser detector 13-5
It is mounted in the microchannel right above deposit chamber, deposits width of the width greater than preceding latter linked microchannel of chamber, deposit chamber
I 13-1 of rectangle supplementary structure, II 13-3 of rectangle supplementary structure being spaced apart with cavity inner wall, rectangle are equipped in cavity
II 13-3 of supplementary structure is located at the end of I 13-1 of rectangle supplementary structure and its length is greater than I 13-1's of rectangle supplementary structure
Length, I 13-1 of rectangle supplementary structure include several groups rectangular slab symmetrical above and below and horizontal positioned, phase in same horizontal line
It (is because because what is passed through from deposit chamber goes back in addition to wrapping up cell drop 8 that neighbour two rectangle supplementary structures, I 13-1 is arranged at equal intervals
Have continuous phase liquid, have interval that continuous phase liquid is facilitated to pass through), with respect to the width etc. between two rectangle supplementary structures, I 13-1
Width in front end microchannel, II 13-3 of rectangle supplementary structure include orthogonal one horizontal positioned rectangular slab and one
The rectangular slab being disposed vertically, II 13-3 of rectangle supplementary structure being disposed vertically are located at horizontal positioned rectangle supplementary structure
The end of I 13-1 of rectangle supplementary structure of the top of II 13-3 and II 13-3 of rectangle supplementary structure and top that are disposed vertically
Notch is formed, the package cell drop 8 intracavitary into deposit enters the micro- of rear end from deposit chamber outlet 13-4 after notch extrusion
Channel, the elliptical bead as shown in Figure 2 of package cell drop 8 squeezed out from notch.
Further, a length of 120mm of the PDMS chip 2, width 60mm, a height of 10mm；The width of all microchannels is
0.020mm, a height of 0.010mm.
Further, the drop register I 9, drop register II 10, drop register III 11, drop register IV
12, drop register V 13, in drop register VI 14 deposit chamber cavity length in the horizontal direction be respectively 0.180mm,
0.160mm,0.140mm,0.120mm,0.100mm,0.080mm；The width of each register deposit chamber cavity is 0.040mm, high
It is 0.010mm；The a length of 0.010mm of I 13-1 of rectangle supplementary structure, width 0.003mm, quantity is 8；Rectangle auxiliary knot
II 13-3 of structure a length of 0.020mm, width 0.003mm are divided into same horizontal line between adjacent two rectangles supplementary structure, I 13-1
0.005mm；Width between I 13-1 of rectangle supplementary structure, II 13-3 of rectangle supplementary structure and corresponding register cavity inner wall
Degree is 0.005mm；The end of I 13-1 of rectangle supplementary structure of II 13-3 of rectangle supplementary structure and top that are disposed vertically
The width for forming notch is 0.005mm.
A kind of screening method of the tumour cell screening micro fluidic device based on drop deposit, specific steps are such as
Under:
Step1: injecting fluorinated oil HFE-7500 to continuous phase entrance 4, and the injection of dispersed phase entrance 6 has the blood of tumour cell,
By controlling the entrance velocity of dispersed phase entrance 6 and continuous phase entrance 4 respectively, when normal erythrocyte enters cross copolymerization coke
After structure 5, the package cell drop 8 with single blood erythrocyte by mocro of 0.020mm diameter can be stably generated, when bigger than erythrocyte
Or after small tumour cell enters cross focusing structure 5 altogether, the package cell liquid than having single blood erythrocyte by mocro can be generated
Drip 8 big or small package cell drops 8, package cell drop 8 pass sequentially through drop register I 9, drop register II 10,
Drop register III 11, drop register IV 12, drop register V 13, drop register VI 14, each drop register energy
The quantity for accommodating package cell drop 8 is the packet beyond receiving number that is fixed, finally entering in each drop register
The drop register can be squeezed out a drop of foremost by wrapping up in cell drop 8, the laser detector in each drop register
The number for detecting the package cell drop 8 by respective drop register, and will test result and sends controller to, from last
The package cell drop 8 of one drop register outflow flow to outside from outlet 3；
Step2: due to size and the softness difference of erythrocyte and tumour cell, each drop register is squeezing package
Required number is different when the package cell drop 8 of different cells, is stored in controller all normal in the set time
By the quantity of the package cell drop 8 of each drop register when erythrocyte passes through, controller is according to each drop register
In the real-time detection structure of laser detector judge whether contain tumour cell in blood, in the same time, when by each
Pass through the packet of each drop register when the quantity of the package cell drop 8 of drop register and all normal erythrocytes pass through
The quantity for wrapping up in cell drop 8 is different, then it is assumed that there are tumour cells.
Drop register I 9, drop register II 10, drop register III 11, drop register IV 12, drop register
V 13, the quantity that drop register VI 14 can accommodate the package cell drop 8 with single blood erythrocyte by mocro is respectively 7,6,5
A, 4,3,2.
The working principle of the invention is: due to the size and pliability difference of tumour cell and haemocyte, being posted by drop
Storage realizes that the frequency passed through to the swollen tumour cell drop of package controls, the frequency passed through using laser detection drop, from
And tumour cell is detected whether.
Drop register I 9, drop register II 10, drop register III 11, drop register IV 12 in the present invention,
Drop register V 13, drop register VI 14 are able to accommodate the package cell that 7 to 2 diameters are 0.020mm simultaneously
Drop 8, can be a package cell of foremost after the 8th package cell drop 8 enters as shown in drop register I 9
Drop 8 squeezes out drop register I 9, such as the package cell drop 8 that the elliptical bead in Fig. 2 will be videlicet squeezed away, due to
The size and softness difference, each register of erythrocyte and tumour cell wrap up the drop when institute of different cells squeezing
The number needed is different, to judge whether contain tumour cell in blood.It wraps up cell drop 8 and exports 13-4 from register
By laser detection 13-5 after out, to obtain the frequency that passes through of package cell drop 8, package cell drop 8 out according to
It is secondary to pass through drop register II 10, drop register III 11, drop register IV 12, drop register V 13, drop register
VI 14, principle is as drop register I 9, but the amount of droplets that register accommodates is held according to package cell size is different
The number received is different.
The length of normal erythrocyte is probably in 0.008mm or so, and tumour cell is not of uniform size, has bigger than erythrocyte
, also have smaller than erythrocyte, it is assumed that the drop of normal package erythrocyte passes through drop register I 9, drop register
II 10, drop register III 11, drop register IV 12, drop register V 13, drop register VI 14 extrude needs
Quantity is respectively 8,7,6,5,4,3, squeezes out a drop register I 9, drop register II 10, drop register III 11, drop
Register IV 12, drop register V 13, drop register VI 14 need time be 1s, a package cell drop 8 from into
Enter to the time gone out and probably needs 33s；If the package cell drop 8 of one times smaller than erythrocyte or so tumour cell is A, than
The package cell drop 8 that erythrocyte is twice left and right tumour cell is B, when including one in the package cell drop 8 of generation
A cell A or B, each of available following table send the number that device passes through:
After detection, each register is grouped together into the intrinsic of this cell by wrapping up the number of cell drop 8
Feature combination, it can be deduced that the content of Fig. 3, so as to judge whether to have A B tumour cell, other sizes it is swollen
Oncocyte testing principle is the same, can shorten detection time in this way, realizes accurate detection.
Drop microflow control technique has become the mainstream scientific research of microflow control technique in recent years with its high throughput, fireballing feature
Hot spot, it biologic grain accurate quantification and the unique advantage of quick context of detection.The present invention is with drop microflow control technique
Point is sent out, the key matter of science and technology such as high-throughput, quick detection towards biologic grain conducts a research, thin to tumour to realize
The accurate detection of the biological samples such as born of the same parents provides theoretical foundation and technical support for early diagnosis, the quick treatment etc. of disease.It is this
New method will help doctor preferably to carry out diagnosing and treating.
The present invention, which realizes, is integrated in sampling fluids, driving, manipulation, transport and detection and analysis on micro-fluidic chip, real
Existing liquid flow it is controllable, micro and trace materials transport and efficient detection, manufacturing cost is low, easy to carry, consumption reagent
Seldom, analysis speed is fast, detection efficiency is high.
In conjunction with attached drawing, the embodiment of the present invention is explained in detail above, but the present invention is not limited to above-mentioned
Embodiment within the knowledge of a person skilled in the art can also be before not departing from present inventive concept
Put that various changes can be made.
Claims (7)
1. a kind of tumour cell screening micro fluidic device based on drop deposit, it is characterised in that: including glass substrate (1) and admittedly
It is scheduled on PDMS chip (2), controller thereon, continuous phase entrance (4) is contained on PDMS chip (2), focusing structure (5), is connected
Continuous mutually outlet (7), dispersed phase entrance (6), register channel, outlet (3), focusing structure (5) are crisscrossing channels, focusing structure
(5) two entrances end is connect with continuous phase entrance (4), dispersed phase entrance (6) respectively by microchannel, and two outlet ends pass through
Microchannel connect with continuous phase outlet (7) and the entrance in register channel respectively, the dispersed phase that enters from dispersed phase entrance (6) and
The continuous phase entered from continuous phase entrance (4) generates package cell drop (8) by focusing structure (5) altogether, wraps up cell drop
(8) register channel is entered by the microchannel between focusing structure (5) and the entrance in register channel, register channel be to
Upper continuous bending shape and the register that several structures are identical, length gradually shortens, register are successively arranged in it from front to back
It is connect with controller, two neighboring register is staggeredly located at the bending part in register channel up and down, wraps up cell drop (8) with solid
Fixed frequency passes sequentially through each register, the number of the detectable package cell drop (8) by it of each register, deposit
The outlet in device channel is connect by microchannel with outlet (3).
2. a kind of tumour cell screening micro fluidic device based on drop deposit according to claim 1, it is characterised in that:
The outlet (3), continuous phase entrance (4), dispersed phase entrance (6), continuous phase outlet (7) proper alignment are in PDMS chip (2)
Left side, register channel be arranged PDMS chip (2) right side.
3. a kind of tumour cell screening micro fluidic device based on drop deposit according to claim 1, it is characterised in that:
Drop register I (9), drop register II (10), drop register are successively arranged in the register channel from front to back
III (11), drop register IV (12), drop register V (13), drop register VI (14), between adjacent drops register
Connected by microchannel, drop register V (13) include deposit chamber, laser detector (13-5), laser detector (13-5) with
Controller connection, deposit chamber include deposit chamber entrance (13-2), cavity, deposit chamber outlet (13-4), are deposited chamber entrance (13-2)
It is connect with front end microchannel, deposit chamber outlet (13-4) is connect with the vertical microchannel of bending part, laser detector (13-5) peace
In the microchannel right above deposit chamber, the width for depositing chamber is greater than the width of preceding latter linked microchannel, deposits the chamber of chamber
It is equipped with rectangle supplementary structure I (13-1), the rectangle supplementary structure II (13-3) being spaced apart with cavity inner wall in vivo, it is rectangular
Shape supplementary structure II (13-3) is located at the end of rectangle supplementary structure I (13-1) and its length is greater than rectangle supplementary structure I
The length of (13-1), rectangle supplementary structure I (13-1) includes several groups rectangular slab symmetrical above and below and horizontal positioned, same
Adjacent two rectangles supplementary structure I (13-1) is arranged at equal intervals on horizontal line, between opposite two rectangle supplementary structures I (13-1)
Width be equal to the width of front end microchannel, rectangle supplementary structure II (13-3) includes orthogonal one horizontal positioned length
Square plate and the rectangular slab that is disposed vertically, the rectangle supplementary structure II (13-3) being disposed vertically are located at horizontal positioned length
The top of rectangular supplementary structure II (13-3) and the rectangle supplementary structure II (13-3) being disposed vertically and the rectangle of top are auxiliary
The end of structure I (13-1) is helped to form notch, the package cell drop (8) intracavitary into deposit is after notch extrusion from deposit chamber
Export the microchannel that (13-4) enters rear end.
4. a kind of tumour cell screening micro fluidic device based on drop deposit according to claim 1 or 3, feature exist
In: a length of 120mm of the PDMS chip (2), width 60mm, a height of 10mm；The width of all microchannels is 0.020mm, a height of
0.010mm。
5. a kind of tumour cell screening micro fluidic device based on drop deposit according to claim 3, it is characterised in that:
The drop register I (9), drop register II (10), drop register III (11), drop register IV (12), drop are posted
In storage V (13), drop register VI (14) deposit chamber cavity length in the horizontal direction be respectively 0.180mm, 0.160mm,
0.140mm,0.120mm,0.100mm,0.080mm；The width of each register deposit chamber cavity is 0.040mm, Gao Junwei
0.010mm；Rectangle supplementary structure I (13-1) a length of 0.010mm, width 0.003mm, quantity are 8；Rectangle supplementary structure
II (13-3) a length of 0.020mm, width 0.003mm, the interval of adjacent two rectangles supplementary structure I (13-1) in same horizontal line
For 0.005mm；Rectangle supplementary structure I (13-1), rectangle supplementary structure II (13-3) and corresponding register cavity inner wall it
Between width be 0.005mm；The rectangle supplementary structure I of the rectangle supplementary structure II (13-3) and top that are disposed vertically
The width that the end of (13-1) forms notch is 0.005mm.
6. a kind of screening side of the described in any item tumour cell screening micro fluidic devices based on drop deposit of claim 1-5
Method, it is characterised in that: specific step is as follows:
Step1: fluorinated oil HFE-7500 is injected to continuous phase entrance (4), dispersed phase entrance (6) injection is with tumour cell
Blood, by controlling the entrance velocity of dispersed phase entrance (6) and continuous phase entrance (4) respectively, when normal erythrocyte enters ten
After font is total to focusing structure (5), the package cell drop (8) with single blood erythrocyte by mocro of 0.020mm diameter can be stably generated,
After tumour cell bigger than erythrocyte or small enters cross focusing structure (5) altogether, it can generate than with single blood red
The big or small package cell drop (8) of the package cell drop (8) of cell, package cell drop (8) pass sequentially through drop and post
Storage I (9), drop register II (10), drop register III (11), drop register IV (12), drop register V (13),
Drop register VI (14), the quantity that each drop register can accommodate package cell drop (8) is fixed, each drop
What is finally entered in register can squeeze out the drop a drop of foremost beyond the package cell drop (8) for accommodating number
Register, the laser detector in each drop register detect the package cell drop (8) by respective drop register
Number, and will test result and send controller to, from the package cell drop (8) that the last one drop register flows out from going out
Mouth (3) flow to outside；
Step2: due to size and the softness difference of erythrocyte and tumour cell, each drop register is squeezing package
Required number is different when the package cell drop (8) of different cells, is stored in the set time in controller and is all positive
By the quantity of the package cell drop (8) of each drop register when normal erythrocyte passes through, controller is posted according to each drop
The real-time detection structure of laser detector in storage judges whether contain tumour cell in blood, in the same time, when logical
It crosses when the quantity of the package cell drop (8) of each drop register passes through with all normal erythrocytes and is deposited by each drop
The quantity of the package cell drop (8) of device is different, then it is assumed that there are tumour cells.
7. the screening method of the tumour cell screening micro fluidic device as claimed in claim 6 based on drop deposit, feature exist
In: drop register I (9), drop register II (10), drop register III (11), drop register IV (12), drop deposit
The quantity that device V (13), drop register VI (14) can accommodate the package cell drop (8) with single blood erythrocyte by mocro is respectively 7
A, 6,5,4,3,2.
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