CN103977468B - For separating of removing circulating tumor cell and hematoblastic system and method in blood - Google Patents

For separating of removing circulating tumor cell and hematoblastic system and method in blood Download PDF

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CN103977468B
CN103977468B CN201410225082.XA CN201410225082A CN103977468B CN 103977468 B CN103977468 B CN 103977468B CN 201410225082 A CN201410225082 A CN 201410225082A CN 103977468 B CN103977468 B CN 103977468B
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blood
micro
separating
tumor cell
circulating tumor
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CN103977468A (en
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蒋兴宇
逯文晶
孙佳姝
王纪东
张璐
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National Center for Nanosccience and Technology China
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National Center for Nanosccience and Technology China
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/38—Removing constituents from donor blood and storing or returning remainder to body, e.g. for transfusion
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3621—Extra-corporeal blood circuits

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Abstract

The invention discloses for separating of removing circulating tumor cell and hematoblastic system and method in blood, relate to tumour cell and separate and remove technical field, for solve existing piece-rate system separating cycle tumour cell weak effect, can not separate blood platelet, the problem such as recycling that can not realize blood designs. Separate with hematoblastic the first microfluidic separation unit that removal system comprises blood collection device and is connected with blood collection device by pipeline for separating of circulating tumor cell in blood, also comprise the second microfluidic separation unit being connected with the first microfluidic separation unit by pipeline, the first microfluidic separation unit is for separating of the circulating tumor cell in blood; The second microfluidic separation unit is for separating of the blood platelet in blood. Propose for separating of circulating tumor cell in blood and hematoblastic method simultaneously. Piece-rate system of the present invention can effectively be removed circulating tumor cell and the blood platelet in cancer patient's blood, easy and simple to handle, and good separating effect can effectively be controlled the diffusion transfer of cancer cell.

Description

For separating of removing circulating tumor cell and hematoblastic system and method in blood
Technical field
The present invention relates to tumour cell and separate removal technical field, relate in particular to a kind of for separating of removing bloodMiddle circulating tumor cell and hematoblastic system and method.
Background technology
Cancer is one of maximum killer of serious harm human health in recent years, and its death rate causing is in the worldEvery country is all positioned at prostatitis. Along with the development of medical science and technology, cancer diagnosis treatment aspect has obtained certainProgress, but can there is recurrence and transfer in various degree in most of patient. Research in recent years shows allThe recurrence of multifactor impact cancer and transfer, wherein peripheral circulation tumour cell has very important effect.Circulating tumor cell is to be come off and invaded the tumour cell of peripheral blood or Lymphatic Circulation by primary tumo(u)r.
In blood because of cancer mortality patient, find tumour cell in 1 example for the first time from Ashworth in 1869After, about the research of circulating tumor cell in peripheral blood be subject to vast clinical and basic medical research person extensivelyPay close attention to. Recent clinical research shows, in cancer patient's blood, hematoblastic quantity is apparently higher than normallyPeople. Therefore carry out circulating tumor cell and hematoblastic fast high flux screening in peripheral blood, find in the past by groupThe Pathological Information that body average data is covered, contributes to realize early diagnosis and the relapse and metastasis monitoring of tumour,The separation removal of carrying out circulating tumor cell in blood can be oncotherapy provides new target spot and strategy.
The immunomagnetic beads that in existing blood, the separation of circulating tumor cell mainly includes based on immunocapture dividesFrom and the microfiltration technology based on physical characteristic difference, the nano wire capture technique based on micro nano structure etc.At present low, the complex operation step of these method ubiquity capture rates, sensitivity are low, are easy to blood sampleProduct cause the shortcomings such as irreversibility destruction. Owing to blood sample having been caused to irreversibility destruction, therefore existingIsolation technics can only carry out preliminary analyzing and testing to blood, and can not be by whole cancer patient bloodTumour cell is removed, and therefore can not reach result for the treatment of. The research of existing circulating tumor cell also mainly stopsThe separation of staying circulating tumor cell detects to be analyzed, and removes blood for effectively separate removal means as oneCirculating tumor cell researchs whole in liquid have not been reported. Separate with hematoblastic for circulating tumor cellRemoval need the appearance of new technology.
Summary of the invention
An object of the present invention is to propose a kind of easy and simple to handle, good separating effect for separating of removing bloodMiddle circulating tumor cell and hematoblastic system.
A further object of the present invention be propose one avoid to blood cause irreversibility destroy, can be to bloodLiquid re-uses, effectively prevents and prevent the thin for separating of circulating tumor in removal blood of cancer metastasisBorn of the same parents and hematoblastic system.
Also object of the present invention be propose a kind of easy and simple to handle, good separating effect for separating of removing bloodCirculating tumor cell and hematoblastic method in liquid.
For reaching this object, on the one hand, the present invention by the following technical solutions:
For separating of removing circulating tumor cell and hematoblastic system in blood, comprise blood collection device withAnd the first microfluidic separation unit being connected with blood collection device by pipeline, also comprise by pipeline withThe second microfluidic separation unit that described the first microfluidic separation unit is connected, wherein,
The first microfluidic separation unit is for separating of the circulating tumor cell in blood;
The second microfluidic separation unit is for separating of the blood platelet in blood.
Preferably, described the first microfluidic separation unit comprises at least one first micro-fluidic chip, describedTwo microfluidic separation unit comprise arrange corresponding to described at least one first micro-fluidic chip at least one theTwo micro-fluidic chips;
The arrival end of the microchannel of described the first micro-fluidic chip is connected with described blood collection device, and describedThe port of export of the microchannel of one micro-fluidic chip separates three branch outlets, the blood platelet in blood, red blood cellWith leucocyte in the first branch outlet flows into the microchannel of the second micro-fluidic chip corresponding thereto, bloodIn circulating tumor cell flow out through the second branch outlet, other compositions in blood flow through the 3rd branch outletGo out;
The port of export of the microchannel of described the second micro-fluidic chip separates two branch outlets, and blood platelet is through the 4thBranch outlet flows out, and red blood cell and leucocyte flow out through quintafurcation outlet.
Preferably, three of the port of export of described the first micro-fluidic chip microchannel branch outlets and describedThe position of two branch outlets of the port of export of two micro-fluidic chip microchannels is by kinetic model
d υ P ‾ dt = 18 μ ρ P α 2 C D R e s 24 ( υ ‾ - υ P ‾ ) + g ‾ ( ρ P - ρ ) ρ P + 1 2 ρ ρ P d ( υ ‾ - υ P ‾ ) dt + F L 1 6 π α 3 ρ P Determine,
Wherein,Be the velocity of particle in fluid, μ is the dynamic viscosity of fluid, ρPThat particle is closeDegree, α is particle diameter, CDDrag coefficient, ResRelative Reynolds number,The velocity of fluid,ρ is fluid density,Acceleration of gravity, FLInertia force;
Determine according to above-mentioned kinetic model the direction that in blood, each cell flows out at the port of export, thereby determine eachThe position of individual branch outlet;
Preferably, three of the port of export of described the first micro-fluidic chip microchannel branch outlet edges are by chipDirection outside mind-set is followed successively by the first branch outlet, the second branch outlet and the 3rd branch outlet, described secondTwo branch outlets of the port of export of micro-fluidic chip microchannel are along be followed successively by the by the outside direction of chip centerFour branch outlets and quintafurcation outlet.
Preferably, described piece-rate system also comprises blood collection note return device, described the 3rd branch outlet and instituteStating quintafurcation outlet is connected by pipeline and described blood collection note return device.
Preferably, the microchannel of described the first micro-fluidic chip and described the second micro-fluidic chip is all spiralShape;
Preferably, the microchannel of described the first micro-fluidic chip and described the second micro-fluidic chip is all double helixShape;
Preferred, described microchannel is by enclosing backpitch winding escalators in conplane six circle right-handed screws and sixBecome, described right-handed screw and described backpitch cross at center position;
Further preferred, the outmost turns spiral of described the first micro-fluidic chip and described the second micro-fluidic chipDiameter is 2-8cm.
Preferably, the cross sectional shape of the microchannel of described the first micro-fluidic chip and described the second micro-fluidic chipFor rectangle, trapezoidal or triangle;
Preferably, the cross sectional shape of the microchannel of described the first micro-fluidic chip and described the second micro-fluidic chipFor rectangle, and the long 275-325 μ of rectangle m, wide 30-70 μ m.
Preferred, the long 300 μ m of described rectangle, wide 50 μ m.
Preferably, the cross-sectional width of described the first branch outlet is 90-100 μ m, described the second branch outletCross-sectional width be 110-130 μ m, the cross-sectional width of described the 3rd branch outlet is 75-95 μ m;
Preferably, the cross-sectional width of described the first branch outlet is 95 μ m, described the second branch outlet cutFace width degree is 120 μ m, and the cross-sectional width of described the 3rd branch outlet is 85 μ m.
Preferably, described the first microfluidic separation unit comprises 3 to 5 the first micro-fluidic chips;
Preferably, described the first microfluidic separation unit comprises 4 the first micro-fluidic chips.
Preferably, the blood of blood collection device collection through peristaltic pump drive flow into successively described first micro-fluidicSeparative element and described the second microfluidic separation unit.
On the other hand, the present invention is by the following technical solutions:
For separating of removing circulating tumor cell and hematoblastic method in blood, apply above-mentioned system, bloodLiquid harvester gathers blood and blood is passed into the first microfluidic separation unit and carries out dividing of circulating tumor cellFrom, then pass into the second microfluidic separation unit and carry out hematoblastic separation.
Beneficial effect of the present invention is:
(1) provided by the inventionly comprise for separating of removing circulating tumor cell and hematoblastic system in bloodThe blood collection device, the first microfluidic separation unit and the second microfluidic separation list that connect successively by pipelineUnit, passes into the first microfluidic separation unit by circulating tumor by blood collection device by cancer patient's bloodCell separation, then passes into the second microfluidic separation unit blood platelet is separated, and can effectively remove cancer and suffer fromCirculating tumor cell in person's blood and blood platelet, easy and simple to handle, good separating effect, can effectively control cancerThe diffusion transfer of cell;
(2) microfluidic separation unit adopts the microchannel of multiple micro-fluidic chips and micro-fluidic chip to adopt two spiral shellsRevolve structure, greatly improved separation flux;
(3) this system also comprises blood collection note return device, because the present invention is in blood based on inertia forceCell separate, therefore can avoid blood to cause irreversibility to destroy, through circulating tumor cellHealthy blood after separation separates with blood platelet can be collected note return device by blood and be collected and note the Huis' body, realThe now blood of washing to cancer patient, effectively the diffusion transfer of prevention and cure cancer, reaches result for the treatment of.
Brief description of the drawings
Fig. 1 be the embodiment of the present invention one provide for separating of removing circulating tumor cell and blood platelet in bloodThe structural representation of system;
Fig. 2 is the first micro-fluidic chip port of export structural representation that the embodiment of the present invention one provides;
Fig. 3 is circulating tumor cell and hematoblastic flow chart in the embodiment of the present invention one separating blood;
Fig. 4 is the schematic diagram that a pair of cancer patient of the embodiment of the present invention washes blood.
In figure, 1, blood collection device; 2, the first microfluidic separation unit; 21, the first micro-fluidic coreSheet; 211, the first branch outlet; 212, the second branch outlet; 213, the 3rd branch outlet; 3, secondMicrofluidic separation unit; 31, the second micro-fluidic chip; 311, the 4th branch outlet; 312, quintafurcationOutlet; 4, blood is collected note return device; 5, cancer patient; 51, circulating tumor cell; 52, blood platelet.
Detailed description of the invention
Further illustrate technical scheme of the present invention below in conjunction with accompanying drawing and by detailed description of the invention.
Embodiment mono-:
The present embodiment has proposed a kind of for separating of removing circulating tumor cell and hematoblastic system in blood,As shown in Figure 1, this system comprises that blood collection device 1, the first microfluidic separation unit 2 and second are micro-fluidicSeparative element 3. The first microfluidic separation unit 2 comprises that four the first micro-fluidic chips 21, the second are micro-fluidicSeparative element 3 comprises four the second micro-fluidic chips 31. Entering of the microchannel of four the first micro-fluidic chips 21Mouth end is all connected with blood collection device 1. As shown in Figure 2, the microchannel of the first micro-fluidic chip 21The port of export separates three branch outlets, along being followed successively by the first branch outlet by the outside direction of chip center211, the second branch outlet 212 and the 3rd branch outlet 213. First point of four the first micro-fluidic chips 21Outlet 211 connects respectively the arrival ends connection of the microchannel of second micro-fluidic chip 31 corresponding with it. TheThe port of export of the microchannel of two micro-fluidic chips 31 separates two branch outlets, and edge is by outside side of chip centerTo being followed successively by the 4th branch outlet 311 and quintafurcation outlet 312.
Separate flux for improving, the microchannel of the first micro-fluidic chip 21 and the second micro-fluidic chip 31 is allDouble-spiral, microchannel is by forming in conplane six circle right-handed screws and six circle backpitch coilings, justSpiral and backpitch cross at center position. Wherein, the first micro-fluidic chip 21 and the second micro-fluidic chip31 outmost turns screw diameter is all preferably 2-8cm. The first branch outlet 211, the second branch outlet 212 andThe 3rd branch outlet 213 is outwards set gradually by the spiral center of the first micro-fluidic chip. The 4th branchOutlet 311 and quintafurcation outlet 312 are outwards set gradually by the spiral center of the second micro-fluidic chip.
In the present embodiment, drive blood flowing and adjusting blood micro-logical in microchannel by peristaltic pumpFlow velocity in road.
Blood is subject to fluid driving force, flox condition, cell-cell interaction, table while flowing in microchannelThe impact of the factors such as surface tension, therefore, the flow path of the cell of different-diameter in microchannel is different, the direction flowing out in exit is also different. For verifying the go forward side by side optimization of line parameter of this conclusion, doCorresponding simulated experiment. First the synthetic different sizes of preparation (3-15 μ polystyrene microsphere simulation blood sample m)In red blood cell, leucocyte, blood platelet and circulating tumor cell, under different rate of flow of fluids, follow the trail of each thinMovement locus when born of the same parents are flowed out and the separating effect of micro-fluidic chip, and micro-by theoretical model calculation optimizationThe geometric parameter of passage and outlet design. In the present embodiment, the cross sectional shape of microchannel is long 275-325μ m, is preferably 300 μ m, and wide 30-70 μ m is preferably the rectangle of 50 μ m. The first branch outlet 211Cross-sectional width be 90-100 μ m, be preferably 95 μ m, the cross-sectional width of the second branch outlet 212 is110-130 μ m, is preferably 120 μ m, and the cross-sectional width of the 3rd branch outlet 213 is 75-95 μ m, excellentElect 85 μ m as.
Can be by setting up kinetic model
d υ P ‾ dt = 18 μ ρ P α 2 C D R e s 24 ( υ ‾ - υ P ‾ ) + g ‾ ( ρ P - ρ ) ρ P + 1 2 ρ ρ P d ( υ ‾ - υ P ‾ ) dt + F L 1 6 π α 3 ρ P Determine each branchThe position of outlet, wherein,Be the velocity of particle in fluid, μ is the dynamic viscosity of fluid, ρPBe particle density, ρ is fluid density, and α is particle diameter, CDDrag coefficient, ResIt is relative ReynoldsNumber,The velocity of fluid,Acceleration of gravity, FLInertia force. Section 1Relevant to viscous force, Section 2Relevant to gravity, Section 3Relevant to surrounding liquid parcel power, Section 4Relevant to inertia force, whereinBinomial and Section 3 impact are little, can ignore, when therefore the cell particle in blood flows out from outletMovement locus is main relevant with suffered viscous force and inertia force. Inertia force is by formula FL＝fL(Re，xL)·ρU2α4/Dh 2Determine, wherein, fL(Re，xL) be at tube section with Reynolds number and particleThe relevant function in position, U is the maximal rate in microchannel, DhFor the hydrodynamics diameter of microchannel.Viscous force FDBy formula FD～ρU2αDh 2/ R is definite, wherein, and the radius of curvature that R is microchannel. By above-mentionedTwo formula known, the diameter of particle is larger, suffered inertia force and viscous force are larger, the change of inertia forceChange degree is greater than the intensity of variation of viscous force, and therefore the diameter of particle is larger, and it more leans in the time that pipe outlet is flowed outNearly centre position, the diameter of particle is less, when it flows out from pipe outlet the closer to chip center, and serum etc.Fluid is not subject to the effect of viscous force, and the outermost exporting at pipe while therefore outflow, away from chip center. ByThis is known, in the time carrying out for the first time cell separation, and blood platelet, red blood cell and thin in vain that in blood, diameter is lessBorn of the same parents in the first branch outlet 211 flows into the microchannel of the second micro-fluidic chip 31 corresponding thereto, diameterLarger circulating tumor cell 51 flows out through the second branch outlet 212, other composition warps such as the serum in bloodThe 3rd branch outlet 213 flows out. In the time carrying out for the second time cell separation, the less blood platelet 52 of diameter is throughFour branch outlets 311 flow out, and the red blood cell that diameter is larger and leucocyte flow out through quintafurcation outlet 312.As shown in fig. 1, dotted portion is the disjunctive path of circulating tumor cell 51 and blood platelet 52, solid line portionBe divided into the disjunctive path of other compositions of blood such as red blood cell, leucocyte and serum.
Because the present embodiment is based on inertia force, the cell in blood to be separated, therefore can avoid bloodLiquid causes irreversibility to destroy, and the 3rd branch outlet 213 can be connected to blood with quintafurcation outlet 312 and receiveFocus return device 4. As shown in Figure 4, cancer patient 5, blood collection device 1, the first microfluidic separation listUnit 2, the second microfluidic separation unit 3 and blood are collected one of the common formation of note return device 4 and are washed blood loop,The blood that blood collection device 1 constantly gathers patient carries out dividing of circulating tumor cell 51 and blood platelet 52From, the healthy blood separating after separating with blood platelet 52 through circulating tumor cell 51 is collected note by bloodReturn device 4 is collected and is noted back in cancer patient's 5 bodies, through constantly circulation, and can be by whole patient bloodIn circulating tumor cell separate and remove, realize the blood of washing to cancer patient 5, reach result for the treatment of. CirculationTumour cell is the approach of cancer metastasis, and the piece-rate system that the present embodiment provides can be by blood samples of patientsCirculating tumor cell is all removed, and can effectively prevent and prevent the diffusion transfer of cancer.
Due to factors such as fluid driving force, flox condition, cell-cell interaction, surface tension in microchannelAll directly affect the behavior of cell in microchannel, the quadratic equation eddying field that double helix micro-structural produces will cause cellMotion in passage is more complicated, presents flow field--the process that intercouples between cell interface develops. CauseThis, application FLUENT carries out Three-dimensional simulation, predicts exactly the mobile row of cell in complex passages systemFor, provide theoretical foundation for optimizing chip design.
The material of the first micro-fluidic chip 21 and the second micro-fluidic chip 31 and manufacturing process and conventional micro-Fluidic chip is identical. For example, adopt soft etching method to make, first photoetching on monocrystalline silicon piece or glass plateObtain the graphics chip of pattern picture, realize the transfer of pattern and copy forming plane and song by Elastic forming boardMicron on face and the micro-fluidic chip of nano-patterning, more specifically, comprise the following steps:
(1) template preparation: chip template is prepared on silicon chip by SU-8 photoresist, by soft etchingTechnology obtains the 3 D stereo template contrary with channel design.
(2) passage forms: make after template, dimethyl silicone polymer liquid prepolymer is cast on silicon chipAnd solidify at a certain temperature, peel off, form double-stranded microchannel layers.
(3) chip sealing: simultaneously process double helix microchannel layers and glass or silicon base also with plasma beamBonding; Using the dimethyl silicone polymer liquid prepolymer of semi-solid preparation as adhesive, fixing involution core after bakingSheet also connects chip outlet and entrance.
Micro-fluidic chip can be made by PDMS, PMMA, PC, bottom sealing-in substrate can use glass, PMMA,PDMS, PC or silicon chip etc.
Propose a kind of for separating of circulating tumor cell in blood and hematoblastic point for above-mentioned piece-rate systemFrom method. As shown in Figure 3, the method is specially, and blood collection device 1 carries out cancer patient 5 automaticallyBlood sampling, first blood pass into the first microfluidic separation unit 2 and carry out cell separation for the first time, diameter in bloodLess blood platelet, red blood cell and leucocyte flows out through the first branch outlet 211, and the circulation that diameter is larger is swollenOncocyte flows out through the second branch outlet 212, and other compositions such as the serum in blood are through the 3rd branch outlet 213Flow out; Flow into second micro-fluidic point from blood platelet, red blood cell and the leucocyte of the first branch outlet outflow 211Carry out cell separation for the second time from unit 3, the blood platelet that diameter is less flows out through the 4th branch outlet 311,The red blood cell that diameter is larger and leucocyte flow out through quintafurcation outlet 312, flow from the 3rd branch outlet 213Other blood constituents such as the serum going out and the red blood cell and the leucocyte menses that flow out from quintafurcation outlet 312Liquid collection note return device 4 is collected and is noted and gets back in cancer patient's 5 bodies, realizes the blood of washing to cancer patient 5,Circulating tumor cell and blood platelet are removed. This mode can effectively be removed in cancer patient's 5 bloodCirculating tumor cell and blood platelet, easy and simple to handle, good separating effect, can effectively control the expansion of cancer cellLoose transfer. The results showed, the separation rate of the circulating tumor cell of this piece-rate system is up to 96.77%.
Wherein, the micro-fluidic chip of the first microfluidic separation unit and the second microfluidic separation unit is not limited toFour, the microchannel of micro-fluidic chip is also not limited to double-spiral structure, can be according to concrete needed separationFlux is set, and the quantity of the micro-fluidic chip of each separative element is preferably 3-5; The cross section shape of microchannelShape is not limited to rectangle, can be also other shapes such as trapezoidal, triangle.
Know-why of the present invention has below been described in conjunction with specific embodiments. These are described just in order to explain thisThe principle of invention, and can not be interpreted as by any way limiting the scope of the invention. Based on hereinExplain, those skilled in the art does not need to pay performing creative labour can associate other tool of the present inventionBody embodiment, within these modes all will fall into protection scope of the present invention.

Claims (16)

1. for separating of removing circulating tumor cell and hematoblastic system in blood, comprise blood collection dressThe first microfluidic separation unit (2) of putting (1) and being connected with blood collection device (1) by pipeline,It is characterized in that: also comprise be connected with described the first microfluidic separation unit (2) by pipeline second micro-Flow Control separative element (3), wherein,
The first microfluidic separation unit (2) is for separating of the circulating tumor cell in blood (51);
The second microfluidic separation unit (3) is for separating of the blood platelet in blood (52);
Described the first microfluidic separation unit (2) comprises at least one first micro-fluidic chip (21), described inThe second microfluidic separation unit (3) comprises corresponding to described at least one first micro-fluidic chip (21) settingAt least one second micro-fluidic chip (31);
The arrival end of the microchannel of described the first micro-fluidic chip (21) and described blood collection device (1) connectConnect, the port of export of the microchannel of described the first micro-fluidic chip (21) separates three branch outlets, in bloodBlood platelet, red blood cell and leucocyte flow into the second miniflow corresponding thereto through the first branch outlet (211)In the microchannel of control chip (31), the circulating tumor cell (51) in blood is through the second branch outlet (212)Flow out, other compositions in blood flow out through the 3rd branch outlet (213);
The port of export of the microchannel of described the second micro-fluidic chip (31) separates two branch outlets, blood platelet(52) flow out through the 4th branch outlet (311), red blood cell and leucocyte are through quintafurcation outlet (312)Flow out.
2. according to claim 1 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: three of the port of export of described the first micro-fluidic chip (21) microchannel branch outThe position of two branch outlets of the port of export of mouth and described the second micro-fluidic chip (31) microchannel is by movingMechanical model
d υ P ‾ d t = 18 μ ρ P α 2 C D Re s 24 ( υ ‾ - υ P ‾ ) + g ‾ ( ρ P - ρ ) ρ P + 1 2 ρ ρ P d ( υ ‾ - υ P ‾ ) d t + F L 1 6 πα 3 ρ P Determine,
Wherein,Be the velocity of particle in fluid, μ is the dynamic viscosity of fluid, ρPIt is particleDensity, α is particle diameter, CDDrag coefficient, ResRelative Reynolds number,The speed that is fluid is vowedAmount, ρ is fluid density,Acceleration of gravity, FLInertia force;
Determine according to above-mentioned kinetic model the direction that in blood, each cell flows out at the port of export, thereby determine eachThe position of individual branch outlet.
3. according to claim 1 for separating of removing circulating tumor cell and hematoblastic system in bloodSystem, is characterized in that: three branch outlets of the port of export of described the first micro-fluidic chip (21) microchannelAlong being followed successively by the first branch outlet (211), the second branch outlet (212) by the outside direction of chip centerWith the 3rd branch outlet (213), two points of the port of export of described the second micro-fluidic chip (31) microchannelExpenditure opening's edge is followed successively by the 4th branch outlet (311) and quintafurcation outlet by the outside direction of chip center(312)。
4. according to claim 1 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: described for separating of also removing in blood circulating tumor cell and hematoblastic systemComprise blood collection note return device (4), described the 3rd branch outlet (213) and the outlet of described quintafurcation(312) collecting note return device (4) by pipeline and described blood is connected.
5. according to claim 1 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: described the first micro-fluidic chip (21) and described the second micro-fluidic chip (31)Microchannel all twist.
6. according to claim 1 for separating of removing circulating tumor cell and hematoblastic system in bloodSystem, is characterized in that: described the first micro-fluidic chip (21) and described the second micro-fluidic chip (31)Microchannel is all Double-spiral.
According to described in claim 5 or 6 for separating of removing circulating tumor cell and blood platelet in bloodSystem, it is characterized in that: described microchannel by conplane six circle right-handed screws and six circle backpitchCoiling forms, and described right-handed screw and described backpitch cross at center position.
8. according to claim 7 for separating of removing circulating tumor cell and hematoblastic system in bloodSystem, is characterized in that: described the first micro-fluidic chip (21) and described the second micro-fluidic chip (31)Outmost turns screw diameter is 2-8cm.
9. according to claim 1 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: described the first micro-fluidic chip (21) and described the second micro-fluidic chip (31)The cross sectional shape of microchannel be rectangle, trapezoidal or triangle.
10. according to claim 9 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: described the first micro-fluidic chip (21) and described the second micro-fluidic chip (31)The cross sectional shape of microchannel be rectangle, and the long 275-325 μ of rectangle m, wide 30-70 μ m.
11. is according to claim 10 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: the long 300 μ m of described rectangle, wide 50 μ m.
12. is according to claim 1 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: the cross-sectional width of described the first branch outlet (211) is 90-100 μ m, described inThe cross-sectional width of the second branch outlet (212) is 110-130 μ m, described the 3rd branch outlet (213)Cross-sectional width is 75-95 μ m.
13. is according to claim 11 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: the cross-sectional width of described the first branch outlet (211) is 95 μ m, described secondThe cross-sectional width of branch outlet (212) is 120 μ m, the cross-sectional width of described the 3rd branch outlet (213)Be 85 μ m.
14. is according to claim 1 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: described the first microfluidic separation unit (2) comprises 3 to 5 the first micro-fluidic coresSheet (21).
15. is according to claim 14 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: described the first microfluidic separation unit (2) comprises 4 the first micro-fluidic chips(21)。
16. is according to claim 1 for separating of removing circulating tumor cell and hematoblastic in bloodSystem, is characterized in that: the blood that blood collection device (1) gathers drives described in inflow successively through peristaltic pumpThe first microfluidic separation unit (2) and described the second microfluidic separation unit (3).
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