CN109580323A - A kind of spiral shape microchannel and its application method and series and parallel mounting structure - Google Patents
A kind of spiral shape microchannel and its application method and series and parallel mounting structure Download PDFInfo
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- 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
- B01L3/502761—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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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Abstract
The invention belongs to rare cells or particle rich to screen field, it is related to a kind of spiral shape microchannel and its application method and string, it is installed in parallel structure, technical problem for spiral shape microchannel in the sorting enrichment field of Peripheral Blood from Patients with Malignant circulating tumor cell, dexterously have devised the series connection and parallel spiral shape microchannel inertial separation structure of the multiple spiral shape microchannels of reasonable application of the manystage cascade connection, relative to existing similar technique, the present invention is enriched with Peripheral Circulation tumour cell to sorting, not only with the capture rate and processing flux of good circulating tumor cell, the purity of circulating tumor cell can also be maximized, minimize the loss of circulating tumor cell, and maintain the original phenotype of circulating tumor cell, cell suspension obtained containing high-purity circulating tumor cell is particularly suitable for subsequent biomedical detection, it loses Credit analysis, cell culture, xenograft tumor preparation etc. are passed, there is extensive and far-reaching clinical practice application value.
Description
Technical field
The invention belongs to rare cell or particle rich to screen field, be related to a kind of spiral shape microchannel and its application method
With series and parallel mounting structure.
Background technique
Invasion and transfer are the Major Clinical causes of death of tumor patient, and death caused by metastases disease accounts for entity
Dead 90% or so caused by tumor, therefore, precisely detect that seem excellent be important the early stage of metastases.Circulating tumor cell refers to
It is detached from primary and metastatic tumour lesion, invades and enter the tumour cell of the circulatory systems such as lymphatic vessel, blood.It is examined in the circulatory system
Measure the presence that circulating tumor cell prompt there may be tumor micrometastasis, and the possibility as DISTANT METASTASES IN that makes further progress
Property.In recent years, many research reports have confirmed circulating tumor cell in the early diagnosis of metastases, individualized treatment, spy
Rope mechanism of tumor metastasis etc. has potential important value.Relative to the tumor tissue cell of solid tumor, Peripheral Circulation
The detection of tumour cell has many advantages, such as that repeatable detection, sampling be simple, hurtless measure, referred to as liquid biopsy, in recent years at
The hot spot precisely diagnosed for tumour.
Circulating tumor cell in the circulatory system not only has morphology and science of heredity heterogeneous but also very rare, even if
It is late tumor patient, the circulating tumor cell in every milliliter of whole blood can only have 1-10, therefore, Yao Shixian circulating tumor cell
Detection, it is an essential step that sorting enrichment is carried out to it, also, the superiority and inferiority of circulating tumor cell sorting enrichment will
Directly affect its subsequent detection effect (such as: counting, gene magnification, gene sequencing, cell culture).
Micro-fluidic chip based on inertia miniflow principle becomes the normal of unmarked sorting enrichment cycles tumour cell in recent years
It with one of method, is mainly characterized by without applying any external force, such as electric power, magnetic force, so that it may be realized in microchannel and include
The focusing of certain size particle including cell is flowed.It is divided by chip structure feature, the miniflow based on inertia miniflow principle
Control chip can be broadly divided into 3 kinds of linear microchannel, curved arc shape microchannel, spiral shape microchannel etc., wherein spiral shape microchannel
It is a kind of special shape in curved arc channel.When fluid is laminar state in linear path, one kind is presented in the flow velocity of fluid
Parabolic shape distribution: the fluid interval velocity among channel is maximum, and for fluid layer closer to conduit wall, flow velocity is lower, also, with
Fluid layer is reduced close to the distance of conduit wall, and flow velocity also reduces according to a certain percentage, and the fluid velocity nearest from conduit wall is most
It is small.A kind of shear force gradient, the liter that this shear force gradient induction generates are produced with the velocity flow profile of this parabolic shape
Power can push the particle to suspend in a fluid to conduit wall.When particle migration is to apart from conduit wall position close enough, channel
Particle can be pushed away conduit wall again by the lift that wall induction generates.The resultant force of above two contrary lift is referred to as inertia liter
Power FL.Inertia lift acts on particle, and particle can be made to generate relative movement in channel cross-section.When particle be moved to it is transversal
At this position of cross section, the inertia lift being subject to is zero, and particle will be stablized in cross section for the equilbrium position in face, i.e. particle
This position forms so that particle be made to focus settling position in cross-section and focuses flowing, flow to downstream.Particle is in linear
In microchannel cross-section the position of stress balance and the shape of microchannel cross-section in relation to: in the microchannel of circular cross section,
Particle can focus to form an annular；In the microchannel of square cross section, the position that particle focuses can be reduced to four, point
Not Wei Yu four faces centre；In the microchannel of rectangular cross section, when microchannel it is bigger in length and breadth when, i.e., microchannel is special
When wide or especially high, the position that particle focuses can be reduced to two, be located at two centres compared with long face.
When fluid flows in curved arc shape microchannel, situation is than more complicated in linear microchannel.Parabolically flow
Dynamic fluid in the channel between speed it is maximum.When by microchannel turning, the fluid among microchannel is because of its flow velocity maximum
And the centrifugal force being subject to is maximum, to flow to the lateral wall of curved arc shape microchannel.Fluid flow rate close to microchannel wall is minimum, institute
It is also minimum by centrifugal force, thus the extruding by intermediate high flow rate fluid.In order to keep in fluid the conservation of mass everywhere, vertical
In on the direction of fluid flowing, formed it is a pair of reversely rotate and symmetrical vortex, be located at microchannel cross-section top and
Thus lower part generates a kind of Secondary Flow referred to as Dien vortex.Dien vortex can generate drag force effect to the particle in fluid,
Referred to as Dien drag force FD.Therefore in curved arc shape microchannel, the particle of flowing can be simultaneously by inertia lift and Dien drag force
Effect, the relative size of both power determines the focusing mobility status that particle flows in curved curved channel.
Microchannel spiral shape microchannel is a kind of special curved arc shape microchannel, it had both had the above-mentioned of curved arc shape microchannel
Focus the characteristic of flowing, it may have the exclusive feature of itself, accordingly, with respect to above-mentioned linear and curved arc shape microchannel, spiral shell
Revolve shape microchannel has unique application advantage in inertia micro flow chip.The inertia of laminar fluid moves in spiral shape microchannel
It moves under power and Dien drag force collective effect, particle moves back and forth between insides of pipes wall and lateral wall and forms Dien vortex.
Above-mentioned lateral transfer is referred to as the Dien period: when one initially lateral by the microchannel of certain length in the particle of outer side wall
When being moved to inner sidewall, it is referred to as completing 0.5 Dien period；When this identical particle is micro- using the downstream of certain length
When channel is back to initial lateral wall, being referred to as completing a complete Dien period completes 1 enlightening in other words
The grace period.As described above, biggish particle (especially its diameter reaches a/h >=0.07) can also be simultaneously other than Dien drag force
By the effect of inertia lift, so that the focal position of particle be made to be very dependent on the ratio (F of inertia lift Yu Dien drag forceL/
FD), the size of above-mentioned ratio and particle has exponent relation, and the focusing of particle depends on the smallest microchannel scale parameter
(the height h) of such as spiral shape microchannel.Biggish particle (that is: a/h >=0.07), Dien drag force and inertia lift are in one and put down
Weighing apparatus state can focus on the inner sidewall of spirality channel behind the microchannel of certain length, and begin in the microchannel in downstream
It is maintained at the inner sidewall of microchannel eventually；And smaller particless (that is: a/h < 0.07), Dien drag force are dominant, particle is with dien vertex
Manifold formula is constantly traversing back and forth between the lateral wall and inner sidewall in microchannel.
In spiral shape microchannel, particle entry can be designed by the exterior sidewall portions in microchannel, in microchannel
Sidewall sections design sheath fluid entrance.With this condition, the sheath fluid fluid with fair speed can squeeze particle flux to microchannel
Lateral wall；When particle flows in microchannel, the effect of the particles of all original states (i.e. inlet) in Dien drag force
Under, inner sidewall is moved to through the micro-tunnel length in 0.5 Dien period by lateral wall；After reaching nearly inner sidewall, biggish grain
Sub (that is: a/h >=0.07) brute force focussing is in inner sidewall due to by strong inertia lift, and further in the work of Dien drag force
It prevents it further mobile to the lateral wall of microchannel with lower, therefore, it is flat to reach some in above-mentioned inertia lift and Dien drag force
When weighing apparatus point, biggish particle focuses on the inner sidewall of microchannel always, and remains in subsequent downstream microchannel
The focused flow of inner sidewall in microchannel；But 0.5 Dien period be moved to inner sidewall smaller particless (that is: a/h <
0.07) the inertia lift being subject to it is small, what is be primarily subjected to is Dien drag force, therefore, can Dien vortex under the action of in downstream
Microchannel lateral transfer through 1 Dien cycle length in microchannel is moved to lateral wall, if the microchannel long enough in downstream, compared with
Small particles with Dien eddy currents microchannel wall the lateral transfer back and forth of lateral wall and inner sidewall.As it can be seen that larger particle is always
Spiral shape microchannel inner sidewall in inertia focus a fluid stream, and smaller particless then with Dien eddy currents microchannel outside
Wall and inner sidewall lateral transfer back and forth.When the spiral shape microchannel of design specific length, exit just makes lesser particle
In or near lateral wall, so that it may separate larger particle with smaller particless by outlet appropriate, realize specific dimensions grain
The enrichment of son.
The above-mentioned characteristic of spiral shape microchannel is widely used in point of Peripheral Blood from Patients with Malignant circulating tumor cell
Choosing enrichment.Its cardinal principle is: relative in peripheral blood red blood cell (diameter is about 6-8 μm) and leucocyte (diameter is about
8-12 μm), the size of circulating tumor cell (diameter is about 20-30 μm) is bigger, by limiting the rectangular cross section of aspect ratio,
Under particular fluid flow conditions, Peripheral Circulation tumour cell will flow to cell along the inner sidewall focus layer of spiral shape microchannel
It exports, remaining cell in sample is then along the lateral wall laminar flow in the channel to waste liquid outlet, to realize that circulating tumor is thin
The separation of born of the same parents.Meanwhile also it can make have various sizes of cell from each specific outlet point by further designing multiple outlets
Stream enrichment.
Relative to linear type and curved arc shape inertia micro flow chip, spiral shape inertia micro flow chip is in sorting enrichment cycles tumour
Cell field has unique advantage, and is mainly manifested in and can rapidly process whole blood sample, reaches the speed of 3mL/min.
ButExisting spiral shape inertia micro flow chipMainly its master of the single-screw shape microchannel inertia micro flow chip of single channel form
Wanting drawback is that circulating tumor cell purity obtained is low, is answered so as to cause the subsequent identification of circulating tumor cell and clinical practice
With there are bigger difficulty, practical clinical value is lower.Although existing spiral shape microchannel is in sorting enrichment cycles
Biggish insufficient and relevant technical bottleneck is still had in terms of tumour cell, still, existing result of study prompt, if energy
Enough dexterously " cascade " applies multiple single spiral shape microchannels, can be in the premise for ensuring good prize efficiency and processing flux
Under the conditions of, circulating tumor cell purity in the cell mixture that the sorting greatlyd improve is enriched to makes it meet clinical reality
Border demand.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of spiral shape microchannel and its application methods and series and parallel to pacify
Assembling structure.
In order to achieve the above objectives, the invention provides the following technical scheme:
A kind of spiral shape microchannel, it is single for sorting enrichment rare cell or particle, including N number of single-screw shape microchannel
A single-screw shape microchannel includes entrance, the microchannel connecting with inlet seal and seals with channel far from one end of entrance
The outlet of connection, the entrance are converged by cell entry and sheath fluid entrance, and the outlet is by waste liquid outlet and cell
It converges outlet；The cell outlet of (M-1) a single-screw shape microchannel and the cell entry of m-th single-screw shape microchannel
It is tightly connected into (M-1) a classification cycle point, by the waste liquid outlet of (M-1) a single-screw shape microchannel, m-th single-screw
The sheath fluid entrance of shape microchannel and (M-1) a classification cycle point are constituted at (M-1) a classification cycle；
The waste liquid outlet of (M-1) a single-screw shape microchannel at (M-1) a classification cycle is flat equipped with pressure
Weigh module, and the flow rate of liquid of the cell entry of m-th single-screw shape microchannel is v1, (M-1) a single-screw shape microchannel
The flow rate of liquid of cell entry is v2；The flow rate of liquid of the cell outlet of m-th single-screw shape microchannel is v3, and (M-1) is a
The flow rate of liquid of the cell outlet of single-screw shape microchannel is v4；The liquid flow of the waste liquid outlet of m-th single-screw shape microchannel
Speed is v5, and the flow rate of liquid of the waste liquid outlet of (M-1) a single-screw shape microchannel is v6；
The pressure balance module is used to make the liquid flow direction and the of the cell entry of m-th single-screw shape microchannel
(M-1) liquid flow direction of the cell entry of a single-screw shape microchannel is consistent, and 0.7*v2≤v1≤1.3*v2；Make M
The cell outlet of the liquid flow direction and (M-1) a single-screw shape microchannel of the cell outlet of a single-screw shape microchannel
Liquid flow direction is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Go out the waste liquid of m-th single-screw shape microchannel
The liquid flow direction of the waste liquid outlet of the liquid flow direction and (M-1) a single-screw shape microchannel of mouth is consistent, and 0.7*
v6≤v5≤1.3*v6；
N number of single-screw shape microchannel forms non-cross spiral shape microchannel；
Wherein, the cell entry of first single-screw shape microchannel is used to enter the sample liquid containing rare cell or particle
Body；The cell outlet of n-th single-screw shape microchannel is used to collect the rare cell or particle after sorting enrichment；The sheath fluid enters
Mouth has osmotic pressure for adding, and guarantees rare cell or the complete dilution of particle；1 < M≤N, M, N is positive integer.
Optionally, the pressure balance module is (M-1) a single-screw being arranged at (M-1) a classification cycle
The pressure balance microchannel being tightly connected at the waste liquid outlet of shape microchannel and with the waste liquid outlet；The pressure balance microchannel
It is a equipped with (N-1).
Optionally, the pressure balance microchannel and N number of single-screw shape microchannel are arranged in the same plane.
Optionally, the pressure balance module is (M-1) a single-screw being arranged at (M-1) a classification cycle
Pressure pump at the waste liquid outlet of shape microchannel.
Optionally, the entrance of first single-screw shape microchannel and the outlet of n-th single-screw shape microchannel are located at spiral
The outside of shape microchannel geometry.
Optionally, the spiral shape microchannel forms the ring-type being parallel to each other by N number of single-screw shape microchannel.
Optionally, the single-screw shape microchannel between the spiral shape microchannel and single-screw adjacent thereto and parallel
Horizontal distance between shape microchannel is 0.1~1000 μm.
Optionally, the section of single single-screw shape microchannel is rectangle or trapezoidal.
Optionally, the section of the single-screw shape microchannel is rectangle, the ratio w/h of the width w and height h of the rectangle
=1.2~10.
Optionally, the section of the single-screw shape microchannel is trapezoidal, the trapezoidal close spiral shape microchannel
Side height of the side height of geometric center lower than the geometric center of the trapezoidal separate spiral shape microchannel.
Optionally, the width w of microchannel is 0.1-1000 μm, is highly 0.1-800 μm.
Optionally, N=2, the spiral shape microchannel is by first single-screw shape microchannel and second single-screw shape
Microchannel composition.
Optionally, the entrance of single single-screw shape microchannel and the distance between the outlet of the single-screw shape microchannel is
The 1.7-2.0 Dien period of non-target cell or non-target particles hydrodynamics characteristic, alternatively, 2.7-3.0 Dien period, or
Person, 3.7-4.0 Dien period, alternatively, 4.7-5.0 Dien period, alternatively, 5.7-6.0 Dien period, alternatively, 6.7-
7.0 Dien periods, alternatively, 7.7-8.0 Dien period, alternatively, 8.7-9.0 Dien period, alternatively, 9.7-10.0 enlightening
The grace period.
Optionally, the channel cross-section width maximum width of single-screw shape microchannel is l1, and the single-screw shape is micro- logical
The cell entry in road and/or the width of cell outlet are l2,0.1*l1≤l2≤0.4*l1；The sheath fluid of the single-screw shape microchannel
The width of entrance and/or waste liquid outlet is l3,0.1*l1≤l3≤0.9*l1.
Optionally, N=4, the spiral shape microchannel are micro- by first single-screw shape microchannel, second single-screw shape
Channel, third single-screw shape microchannel and the 4th single-screw shape microchannel composition.
Optionally, the diameter of the rare cell or particle is a, and the height of the microchannel is h, 0.07≤a/h≤
0.5。
Optionally, the rare cell or particle are circulating tumor cell, circulation progenitor cells, circulation fetal cell, cell
Vesica, excretion body or pathogen.
It optionally, further include for sorting the rare cell being enriched to the spirality channel or particle is identified
And/or the detection unit counted；The detection unit is arranged at the cell outlet of n-th single-screw shape microchannel.
Optionally, the detection unit carries out the Characters Identification of rare cell or particle by particular technology means；It is described
Particular technology means be selected from the group that is made of Photobiology, electrobiology, bioacoustics, biomagnetism and any combination thereof.
Optionally, the detection unit carries out the identification of rare cell or particle proterties by the detection to special characteristic,
The special characteristic is selected from by the Terahertz wave spectrum of rare cell or particle, Raman spectrum, impedance, differential impedance and its any
The group that group is combined into.
Optionally, a described the pressure balance microchannel and N number of single-screw shape microchannel (N-1) form non-cross spiral shell
Shape microchannel is revolved, the outlet of one or more pressure balance microchannels is arranged in outside the geometric figure of spiral shape microchannel, leads to
The length for crossing adjustment a pressure balance microchannel (M-1) passes through content with internal diameter to adjust in (M-1) a cell outlet
Flow velocity.
Optionally, the outlet of a described pressure balance microchannel (N-1) is arranged at the geometric graph of spiral shape microchannel
Outside shape.
Optionally, the one or more in N number of cell outlet is arranged in outside the geometric figure of spiral shape microchannel.
Optionally, all cell outlets are arranged at outside the geometric figure of spiral shape microchannel.
Optionally, the one or more in N number of entrance is arranged in outside the geometric figure of spiral shape microchannel.
Optionally, all cell entries are arranged at outside the geometric figure of spiral shape microchannel.
Optionally, all sheath fluid entrances are arranged at outside the geometric figure of spiral shape microchannel.
A kind of application method of spiral shape microchannel, comprising the following steps:
S1: sample liquid is imported from the cell entry of first single-screw shape microchannel, by dilution from first list
The sheath fluid entrance of the sheath fluid entrance of spiral shape microchannel to n-th single-screw shape microchannel imports；
S2: the cell outlet in n-th single-screw shape microchannel collects the fluid rich in rare cell or particle；First
Waste liquid is discharged in the waste liquid outlet of the waste liquid outlet of a single-screw shape microchannel to n-th single-screw shape microchannel.
Optionally, the sample liquid used in step S1 is the blood sample with certain hematid specific volume, or passes through
Haemolysis mode removes the blood sample of a large amount of red blood cells；Hematid specific volume in the blood sample is 0%~70%.
Optionally, the dilution used in step S1 has certain osmotic pressure, is phosphoric acid for protecting haemocyte integrality
Salt buffer, culture serum or physiological saline.
It optionally, further include step S1.1；
S1.1: adjustment pressure balance module makes the liquid flowing side of the cell entry of the m-th single-screw shape microchannel
To consistent with the liquid flow direction of cell entry of (M-1) a single-screw shape microchannel, and 0.7*v2≤v1≤1.3*v2；
Make the liquid flow direction of the cell outlet of m-th single-screw shape microchannel and the cell of (M-1) a single-screw shape microchannel
The liquid flow direction of outlet is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Make m-th single-screw shape microchannel
The liquid flow direction of the waste liquid outlet of the liquid flow direction and (M-1) a single-screw shape microchannel of waste liquid outlet is consistent,
And 0.7*v6≤v5≤1.3*v6.
A kind of structure that is installed in series of spiral shape microchannel, which is characterized in that be arranged in series using P spiral shape microchannel
Mode, the cell outlet of the n-th single-screw shape microchannel of a spiral shape microchannel (Q-1) and the Q spiral shape are micro- logical
First single-screw shape microchannel entrance in road is connected, and forms spiral shape microchannel and is installed in series place, 1 < Q≤P.
Optionally, spiral shape microchannel be installed in series place a spiral shape microchannel (Q-1) n-th single-screw shape
Pressure balance module also is provided at the waste liquid outlet of microchannel.
Optionally, the pressure balance module be with spiral shape microchannel be installed in series place (Q-1) a spiral shape it is micro-
The pressure balance microchannel that the waste liquid outlet of the n-th single-screw shape microchannel in channel is tightly connected.
Optionally, the pressure balance microchannel and matched spiral shape microchannel form non-cross spiral shell
Shape microchannel is revolved, the outlet of one or more pressure balance microchannels is arranged in outside the geometric figure of spiral shape microchannel, leads to
The length of toning seamless dynamic balance microchannel adjusts the flow velocity in matched cell outlet by content with internal diameter.
Optionally, the outlet of all pressure balance microchannels is arranged at outside the geometric figure of spiral shape microchannel.
Optionally, the one or more in the cell outlet is arranged in outside the geometric figure of spiral shape microchannel.
Optionally, all cell outlets are arranged at outside the geometric figure of spiral shape microchannel.
Optionally, the one or more in the entrance is arranged in outside the geometric figure of spiral shape microchannel.
Optionally, all cell entries are arranged at outside the geometric figure of spiral shape microchannel.
Optionally, all sheath fluid entrances are arranged at outside the geometric figure of spiral shape microchannel.
Optionally, the pressure balance module is that spiral shape microchannel is arranged in be installed in series (Q-1) a spiral at place
Pressure pump at the waste liquid outlet of the n-th single-screw shape microchannel of shape microchannel.
Optionally, N=2, the spiral shape microchannel are Double-spiral microchannel.
Optionally, P=2, in such a way that 2 spiral shape microchannels are arranged in series, the N of first spiral shape microchannel
The cell outlet of a single-screw shape microchannel is connected with first single-screw shape microchannel entrance of second spiral shape microchannel
It connects.
A kind of spiral shape microchannel is installed in parallel structure, in such a way that X spiral shape microchannel is arranged in parallel, sample
Solution enters from the cell entry of first single-screw shape microchannel of X spiral shape microchannel, and rare cell or particle are from X
The cell outlet of the n-th single-screw shape microchannel of spiral shape microchannel flows out；X > 1.
Optionally, the cell entry of first single-screw shape microchannel of X spiral shape microchannel is interconnected.
Optionally, the cell outlet of the n-th single-screw shape microchannel of X spiral shape microchannel is interconnected.
Optionally, the X spiral shape microchannel is arranged in the same plane.
Optionally, the X spiral shape microchannel is arranged concentrically in the same plane.
Optionally, the X spiral shape microchannel is with one heart and along perpendicular to single spiral shape microchannel place plane
Direction setting.
Optionally, X=2, in such a way that two spiral shape microchannels are arranged in parallel, sample solution is micro- from two spiral shapes
The cell entry of first single-screw shape microchannel in channel enters, rare cell or particle from two spiral shape microchannels the
The cell outlet of N number of single-screw shape microchannel flows out.
Optionally, N=2, in such a way that X Double-spiral microchannel is arranged in parallel, sample solution is from X Double-spiral
The cell entry of first single-screw shape microchannel of microchannel enters, and rare cell or particle are from X Double-spiral microchannel
Second single-screw shape microchannel cell outlet outflow.
Optionally, the waste liquid of the n-th single-screw shape microchannel of the X spiral shape microchannel in spiral shape microchannel
Exit is equipped with pressure balance module.
Optionally, the pressure balance module is pressure pump.
Optionally, the pressure balance module is pressure balance microchannel.
Optionally, the pressure balance microchannel and matched spiral shape microchannel form non-cross spiral shell
Shape microchannel is revolved, the outlet of one or more pressure balance microchannels is arranged in outside the geometric figure of spiral shape microchannel, leads to
The length of toning seamless dynamic balance microchannel adjusts the flow velocity in matched cell outlet by content with internal diameter.
Optionally, the outlet of all pressure balance microchannels is arranged at outside the geometric figure of spiral shape microchannel.
Optionally, the one or more in the cell outlet is arranged in outside the geometric figure of spiral shape microchannel.
Optionally, all cell outlets are arranged at outside the geometric figure of spiral shape microchannel.
Optionally, the one or more in the entrance is arranged in outside the geometric figure of spiral shape microchannel.
Optionally, all cell entries are arranged at outside the geometric figure of spiral shape microchannel.
Optionally, all sheath fluid entrances are arranged at outside the geometric figure of spiral shape microchannel.
A kind of combination connecting installation structure in spiral shape microchannel road, including K microchannel monomer, the microchannel monomer
For spiral shape microchannel described in any one of claim 1~27；The microchannel monomer is using series, parallel, repeatedly string
Connection, type of attachment repeatedly in parallel or combining in series and parallel are combined with each other, and wherein K is positive integer and K >=2；Concatenated type of attachment
Connect to seal first cell entry of the n-th cell outlet of (K-1) a microchannel monomer and k-th microchannel monomer
It connects, forms tandem spiral shape microchannel；Type of attachment in parallel is to enter first cell of (K-1) a microchannel monomer
First cell entry of mouth and k-th microchannel monomer is tightly connected convergence and is combined into unified cell entry；By (K-1)
First sheath fluid entrance of a microchannel monomer is combined with the connection convergence of first sheath fluid inlet seal of k-th microchannel monomer
At unified sheath fluid entrance, parallel spiral shape microchannel is formed.
Optionally, repeatedly concatenated type of attachment is by multiple tandem spiral shapes microchannel with concatenated type of attachment phase
Mutually combination；Or tandem spiral shape microchannel and microchannel monomer are combined with each other with concatenated type of attachment.
Optionally, type of attachment in parallel further includes by the n-th cell outlet and K of (K-1) a microchannel monomer
The n-th cell outlet of a microchannel monomer is tightly connected convergence and is combined into unified cell outlet；By a microchannel (K-1)
The n-th waste liquid outlet of monomer and the n-th waste liquid outlet of k-th microchannel monomer are tightly connected convergence and are combined into unified give up
Liquid outlet.
Optionally, repeatedly the type of attachment of parallel connection is by multiple parallel spiral shape microchannels with type of attachment phase in parallel
Mutually combination；Or parallel spiral shape microchannel is combined with each other with microchannel monomer in form in parallel.
Optionally, the type of attachment combined in series and parallel is by one or more parallel spiral shape microchannels and/or one
Or multiple tandem spiral shapes microchannel and/or one or more microchannel monomers are with concatenated type of attachment or the connection of parallel connection
Form is combined with each other, and forms series parallel type spiral flow channel.
A kind of single-screw shape microchannel, including entrance, the microchannel being connect with inlet seal and with channel far from entrance
The outlet that is tightly connected of one end, the entrance converges by cell entry and sheath fluid entrance, and the outlet is by waste liquid
Outlet and cell outlet converge, wherein cell entry is used to enter the sample liquid containing rare cell or particle；Cell goes out
Mouth is for the rare cell or particle after collecting sorting enrichment；Sheath fluid entrance has osmotic pressure for adding, and guarantees rare cell
Or the complete dilution of particle.
Optionally, the cell entry and/or the setting of sheath fluid entrance and/or waste liquid outlet and/or cell outlet are micro- logical
On the outside of the geometric shape in road.
A kind of structure that is installed in series of single-screw shape microchannel, in such a way that P single-screw shape microchannel is arranged in series,
The cell outlet of (Q-1) a single-screw shape microchannel is connected with the cell entry of the Q single-screw shape microchannel, and 1 < Q≤
P。
Optionally, pressure balance module is equipped at the waste liquid outlet of (Q-1) a single-screw shape microchannel.
Optionally, the pressure balance module is pressure pump or pressure balance microchannel.
Optionally, the pressure balance module is to be tightly connected with the waste liquid outlet of the single spiral shape microchannel (Q-1)
Pressure balance microchannel.
Optionally, the pressure balance microchannel and matched all single-screw shapes microchannel, which are formed, does not hand over mutually
The outlet of the spiral shape microchannel of fork, one or more pressure balance microchannels is arranged in outside the geometric figure of spiral shape microchannel
Portion passes through content by adjusting the length and internal diameter of pressure balance microchannel to adjust in matched cell outlet
Flow velocity.
Optionally, the outlet of all pressure balance microchannels is arranged at outside the geometric figure of spiral shape microchannel.
Optionally, the cell entry and/or sheath fluid entrance and/or waste liquid outlet and/or cell outlet are arranged in spiral
On the outside of the geometric shape of shape microchannel.
A kind of single-screw shape microchannel is installed in parallel structure, in such a way that X single-screw shape microchannel is arranged in parallel,
Sample solution enters from the cell entry of X single-screw shape microchannel, and rare cell or particle are from X single-screw shape microchannel
Cell outlet outflow；X > 1.
Optionally, the cell entry of X single-screw shape microchannel and/or cell outlet are interconnected.
Optionally, pressure balance module is equipped at the waste liquid outlet of X single-screw shape microchannel；The pressure balance
Module is pressure pump or pressure balance microchannel.
Optionally, the pressure balance microchannel and matched single-screw shape microchannel are formed non-cross
The outlet of spiral shape microchannel, one or more pressure balance microchannels is arranged in outside the geometric figure of spiral shape microchannel,
The stream in matched cell outlet by content is adjusted with internal diameter by adjusting the length of pressure balance microchannel
Speed.
Optionally, all cell entries and/or the setting of sheath fluid entrance and/or waste liquid outlet and/or cell outlet are micro- logical
On the outside of the geometric shape in road.
Optionally, all cell entries and/or sheath fluid entrance are arranged at outside the geometric figure of spiral shape microchannel.
The beneficial effects of the present invention are:
Sorting of the present invention for spiral shape microchannel in Peripheral Blood from Patients with Malignant circulating tumor cell is enriched with field
Technical problem, develop more simple and effective technology, dexterously have devised the multiple spiral shape microchannels of reasonable application of the manystage cascade connection
Series connection and parallel spiral shape microchannel inertial separation structure, the cascade realized on individual spiral shape inertia micro flow chip are folded
Add formula classification enrichment and/or purification cycle tumour cell.Relative to existing similar technique, the present invention to sorting enrichment and/or
Peripheral Circulation tumour cell, not only with the capture rate and processing flux of good circulating tumor cell, it is also possible to
The purity for maximizing circulating tumor cell, minimizes the loss of circulating tumor cell, and maintain the original table of circulating tumor cell
Type greatly reduces testing cost, shortens processing time and process, obtained to contain the thin of high-purity circulating tumor cell
Born of the same parents' suspension has particularly suitable for subsequent biomedical detection, genetic analysis, cell culture, xenograft tumor preparation etc.
Extensive and far-reaching clinical practice application value.
Other advantages, target and feature of the invention will be illustrated in the following description to a certain extent, and
And to a certain extent, based on will be apparent to those skilled in the art to investigating hereafter, Huo Zheke
To be instructed from the practice of the present invention.Target of the invention and other advantages can be realized by following specification and
It obtains.
Detailed description of the invention
To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention is made below in conjunction with attached drawing excellent
The detailed description of choosing, in which:
Fig. 1 is the structural schematic diagram of embodiment one in the present invention；
Fig. 2 is the structural schematic diagram of embodiment two in the present invention；
Fig. 3 is the structural schematic diagram of embodiment three in the present invention；
Fig. 4 is the structural schematic diagram of example IV in the present invention；
Fig. 5 is the structural schematic diagram of embodiment five in the present invention；
Fig. 6 is the structural schematic diagram of embodiment six in the present invention；
Fig. 7 is the structural schematic diagram of embodiment seven in the present invention；
Fig. 8 is motion profile figure of the various sizes of particle under different in flow rate in the embodiment of the present invention one；
Fig. 8 a is the motion profile of various sizes of particle that flow velocity is 150 μ l/min in example 1；
Fig. 8 b is the motion profile of various sizes of particle that flow velocity is 300 μ l/min in example 1；
Fig. 8 c is the motion profile of various sizes of particle that flow velocity is 450 μ l/min in example 1；
Fig. 8 d is the motion profile of various sizes of particle that flow velocity is 600 μ l/min in example 1；
Fig. 8 e is the motion profile of various sizes of particle that flow velocity is 750 μ l/min in example 1；
Fig. 8 f is the motion profile of various sizes of particle that flow velocity is 900 μ l/min in example 1；
Fig. 9 is the microscope photograph in different spiral shape microchannels exit in the embodiment of the present invention one；
Fig. 9 a is the displaing micro picture at traditional single-screw shape microchannel cell outlet；
Fig. 9 b is the displaing micro picture at the cell outlet of Double-spiral microchannel of the present invention；
Fig. 9 c is the displaing micro picture at Double-spiral microchannel of the present invention waste liquid outlet 7；
Figure 10 is that fluid-flow rate marks in spiral shape microchannel in the embodiment of the present invention one；
Figure 11 is traditional single-screw shape microchannel；
Figure 12 is the structural schematic diagram of embodiment eight in the present invention；
Figure 13 is the structural schematic diagram of embodiment nine in the present invention；
Figure 14 is the structural schematic diagram of embodiment ten in the present invention；
Figure 15 is the structural schematic diagram of embodiment 11 in the present invention；
Figure 16 is the structural schematic diagram of embodiment 12 in the present invention；
Figure 17 is the structural schematic diagram of embodiment 13 in the present invention；
Figure 18 is the structural schematic diagram of embodiment 14 in the present invention；
Figure 19 is the structural schematic diagram of embodiment 15 in the present invention；
Figure 20 is the structural schematic diagram of embodiment 16 in the present invention；
Figure 21 is the structural schematic diagram of embodiment 17 in the present invention；
Figure 22 is the structural schematic diagram of embodiment 18 in the present invention；
Figure 23 is the simulation result of embodiment eight in the present invention, indicates the movement rail of lower polystyrene microsphere different in flow rate
Mark；
Flow velocity is 200ul/min in Figure 23 a；
Flow velocity is 350ul/min in Figure 23 b；
Flow velocity is 500ul/min in Figure 23 c；
Flow velocity is 650ul/min in Figure 23 d；
Flow velocity is 800ul/min in Figure 23 e；
Flow velocity is 950ul/min in Figure 23 f；
Figure 24 is the simulation result comparison diagram of embodiment eight and embodiment 14 in the present invention；
Figure 24 a indicates the enrichment condition in embodiment eight at the cell outlet of second single-screw shape microchannel；
Figure 24 b indicates the enrichment condition at the cell outlet of single-screw shape microchannel traditional shown in Figure 11；
Figure 24 c indicates the enrichment condition in embodiment eight at the cell outlet of first single-screw shape microchannel；
Figure 24 d indicates the enrichment condition at the waste liquid outlet of single-screw shape microchannel traditional shown in Figure 11；
Figure 24 e indicates the enrichment condition in embodiment eight at the waste liquid outlet of second single-screw shape microchannel；
It is collected at the waste liquid outlet and cell outlet of second single-screw shape microchannel in Figure 25 expression embodiment eight
The immunofluorescence dyeing result of cell；
Figure 25 a is the cell of the cell that is collected under 40 times of light microscopic at the waste liquid outlet of second single-screw shape microchannel
Form；
Figure 25 b is the leucocyte DAPI fluorescent staining result that double helix die grading is enriched to；
Figure 25 c is the leucocyte CD45 antibody fluorescence coloration result that double helix die grading is enriched to；
Figure 25 d is the leucocyte fluorescent staining Merge result that double helix die grading is enriched to；
Figure 25 e is the cell of the cell that is collected under 40 times of light microscopic at the cell outlet of second single-screw shape microchannel
Form；
Figure 25 f is the SW480DAPI fluorescent staining result that double helix die grading is enriched to
Figure 25 g is the SW480EPCAM fluorescent staining result that double helix die grading is enriched to
Figure 25 h is the SW480 fluorescent staining Merge result that double helix die grading is enriched to；
Figure 26 is analysis result of the double helix chip to the capture rate of tumour cell；
Figure 26 a is double helix chip to various concentration tumour cell recovering state in whole blood sample；
Figure 26 b is double helix chip to different tumour cell capture rate histograms.
Specific embodiment
Illustrate embodiments of the present invention below by way of specific specific example, those skilled in the art can be by this specification
Other advantages and efficacy of the present invention can be easily understood for disclosed content.The present invention can also pass through in addition different specific realities
The mode of applying is embodied or practiced, the various details in this specification can also based on different viewpoints and application, without departing from
Various modifications or alterations are carried out under spirit of the invention.It should be noted that diagram provided in following embodiment is only to show
Meaning mode illustrates basic conception of the invention, and in the absence of conflict, the feature in following embodiment and embodiment can phase
Mutually combination.
The element numbers please referred in Fig. 1-Figure 26 attached drawing respectively indicate: the cell entry of first single-screw shape microchannel
1, the entrance A of 2, first single-screw shape microchannels of sheath fluid entrance of first single-screw shape microchannel, pressure balance microchannel
3, the sheath fluid entrance of 4, second single-screw shape microchannels of cell outlet of first single-screw shape microchannel 5, first single spiral shells
Revolve the outlet B of 6, first single-screw shape microchannels of waste liquid outlet of shape microchannel, the waste liquid of second single-screw shape microchannel
The outlet C of 8, second single-screw shape microchannels of cell outlet of 7, the second single-screw shape microchannels in outlet, second spiral
First single-screw of 9, second spiral shape microchannels of sheath fluid entrance of first single-screw shape microchannel in shape microchannel
The sheath fluid entrance 11 of second single-screw shape microchannel of 10, second spiral shape microchannels of waste liquid outlet of shape microchannel,
Second of 12, second spiral shape microchannels of cell outlet of second single-screw shape microchannel of two spiral shape microchannels
The cell entry of first single-screw shape microchannel of 13, second spiral shape microchannels of waste liquid outlet of single-screw shape microchannel
14, the sheath fluid entrance 15 of third single-screw shape microchannel, the waste liquid outlet 16 of third single-screw shape microchannel, the 4th list
The sheath fluid entrance 17 of spiral shape microchannel, the cell outlet 18 of the 4th single-screw shape microchannel, the 4th single-screw shape are micro- logical
The waste liquid outlet 19 in road.
Embodiment one
The present invention relates to a kind of spiral shape microchannels to wrap in the present embodiment for sorting enrichment rare cell or particle
Two single-screw shape microchannels are included, which winds cyclization in the same plane and non-crossly
Shape, as shown in Figure 1.
Single single-screw shape microchannel include entrance A, the microchannel being connect with inlet seal and with channel far from enter
The outlet B that one end of mouth is tightly connected, the entrance are converged by cell entry 1 and sheath fluid entrance 2, the outlet B
It is converged by waste liquid outlet 6 and cell outlet 4；
The cell outlet 4 of first single-screw shape microchannel and the cell entry of second single-screw shape microchannel, which seal, to be connected
It is connected into first classification cycle point, by waste liquid outlet 6, second single-screw shape microchannels of first single-screw shape microchannel
Sheath fluid entrance 5 and first classification cycle point are constituted at first classification cycle；
It is micro- logical that the waste liquid outlet 6 of first single-screw shape microchannel at first classification cycle is equipped with pressure balance
The flow rate of liquid in road 3, the cell entry of second single-screw shape microchannel is v1, and the cell of first single-screw shape microchannel enters
The flow rate of liquid of mouth 1 is v2；The flow rate of liquid of the cell outlet 8 of second single-screw shape microchannel is v3, first single-screw
The flow rate of liquid of the cell outlet 4 of shape microchannel is v4；The flow rate of liquid of the waste liquid outlet 7 of second single-screw shape microchannel is
V5, the flow rate of liquid of the waste liquid outlet 6 of first single-screw shape microchannel are v6；
The pressure balance microchannel be used for make second single-screw shape microchannel cell entry liquid flow direction and
The liquid flow direction of the cell entry 1 of first single-screw shape microchannel is consistent, and 0.7*v2≤v1≤1.3*v2；Make second
The liquid flow direction of the cell outlet 8 of a single-screw shape microchannel and the cell outlet 4 of first single-screw shape microchannel
Liquid flow direction is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Go out the waste liquid of second single-screw shape microchannel
The liquid flow direction of the waste liquid outlet 6 of the liquid flow direction and first single-screw shape microchannel of mouth 7 is consistent, and 0.7*v6
≤v5≤1.3*v6；
Wherein, the cell entry 1 of first single-screw shape microchannel is used to enter the sample liquid containing rare cell or particle
Body；The cell outlet 8 of second single-screw shape microchannel is used to collect the rare cell or particle after sorting enrichment；Described first
The sheath fluid entrance 2 of a single-screw shape microchannel and the sheath fluid entrance 5 of second single-screw shape microchannel are for adding with infiltration
Pressure thoroughly guarantees the dilution of rare cell or granule integrity and biological activity.
Using the form of two single-screw shape microchannels, second single-screw shape microchannel can be directly to first single spiral shell
The rare cell or particle that the sorting of rotation shape microchannel is enriched to carry out further sorting enrichment, thus than single single-screw shape
Microchannel captures the more efficient of rare cell or particle, be equivalent to rare cell or particle carried out sorting enrichment twice and
It is micro- to be superior to single-screw shape by purifying for the quadrinomial parameters such as the capture rate of target cell, handling capacity, cell purity, cell activity
Channel.
Double-spiral microchannel of the present invention is by first single-screw shape microchannel and second single-screw shape microchannel
It is directly connected to mode from beginning to end and constitutes an appearance be in parallel and to have the entirety of the Double-spiral microchannel of identical centre point several
What structure, it is characterized in that the entrance A of first single-screw shape microchannel of Double-spiral microchannel and second single-screw shape are micro-
The outlet in channel is respectively positioned on the outside of microchannel main body geometry.
Each single-screw shape microchannel of Double-spiral microchannel of the present invention have cell entry 1, sheath fluid entrance 2,
Cell outlet 4 and waste liquid outlet 6, wherein cell entry 4 and sheath fluid entrance 2 pool the entrance of each single-screw shape microchannel
A, cell outlet 4 and waste liquid outlet 6 pool the outlet B of each single-screw shape microchannel.Cell entry 1 is usually located at both orders
The exterior sidewall portions of spiral shape microchannel entrance, sheath fluid entrance 2 are usually located at the inner sidewall of set single-screw shape microchannel entrance
Part；Cell outlet 4 is usually located at the inner sidewall part of set single-screw shape microchannel outlet, and waste liquid outlet 6 is usually located at both
The exterior sidewall portions of order spiral shape microchannel outlet.
Two single-screw shape microchannels of Double-spiral microchannel of the present invention have same or similar restriction in length and breadth
The rectangle cross sections of ratio can also be lower than the trapezoid cross section of lateral wall height for inner sidewall height, be used for because target cell is thin with non-target
Size difference existing for born of the same parents and only make target cell microchannel inner sidewall formed focused flow, non-target cell is then with Dien vortex-like fashion
The lateral transfer back and forth between microchannel inner sidewall and lateral wall.Further, the width of microchannel is 0.1-1000 μm, depth
It is 0.1-800 μm, or further, the width of microchannel is 100-800 μm, and depth is 50-500 μm, or further, it is micro- logical
The width in road is 600 μm, and depth is 140-180 μm.
Two single-screw shape microchannels of Double-spiral microchannel of the present invention have same or similar length, special
Sign is that the length of the spiral shape microchannel starting point of the 1st and the 2nd single-screw shape microchannel to terminal is non-target cell fluid
The 1.7-2.0 Dien period of mechanical characteristics, alternatively, 2.7-3.0 Dien period, alternatively, 3.7-4.0 Dien period, or
Person, 4.7-5.0 Dien period, alternatively, 5.7-6.0 Dien period, alternatively, 6.7-7.0 Dien period, alternatively, 7.7-
8.0 Dien periods, alternatively, 8.7-9.0 Dien period, alternatively, 9.7-10.0 Dien period.With this condition,
The terminal point of one and second single-screw shape microchannel, non-target cell is along dien vertex stream migration to the outside of spiral shape microchannel
Wall part.
The waste liquid outlet of first single-screw shape microchannel of Double-spiral microchannel of the present invention has pressure balance micro-
Channel 3, the size and shape of the above-mentioned pressure balance microchannel 3 of the present invention are without particular/special requirement, to its aspect ratio, curvature half
Without particular/special requirement, effect is the cell outlet 1 of first single-screw shape microchannel of balance for diameter, width, height and length
Pressure makes the stream of the cell entry of second single-screw shape microchannel or the cell outlet 4 of first single-screw shape microchannel
Speed and direction and the cell entry 1 of first single-screw shape microchannel and the cell outlet 4 of second single-screw shape microchannel
Flow velocity is identical as direction holding or extremely close, meanwhile, make flow velocity and the direction of the waste liquid outlet 6 of first single-screw shape microchannel
And the flow velocity of the waste liquid outlet 7 of second single-screw shape microchannel is identical as direction holding or extremely close.
The horizontal parallel spacing of first and second spiral shape microchannel of Double-spiral microchannel of the present invention is
0.1-1000μm。
The cell entry 1 of first and/or second spiral shape microchannel of Double-spiral microchannel of the present invention is micro-
The width in channel is the 10-40% of microchannel overall width, the waste liquid outlet 6 of first and/or second spiral shape microchannel
Microchannel width be microchannel overall width 40-90%.
Blood sample of the present invention refers to the blood sample with certain hematid specific volume, alternatively, passing through haemolysis side
Formula removes the blood sample of a large amount of red blood cells, alternatively, by other approach well knowns or device obtain without red blood cell or
Only a small amount of or minute quantity red blood cell blood sample.
Sheath fluid of the present invention refers to certain osmotic pressure, and does not destroy any this field of haemocyte integrality
Well known various solution, such as phosphate buffer, culture serum, physiological saline.
Can particle form the size and radius of curvature, particle that inertia focuses flowing with microchannel in curved arc shape microchannel
Size and the factors such as flow velocity have relationship.Existing research shows that the width in channel is greater than the rectangular section channel of height
In, the particle of certain size forms inertia and focuses flowing it is generally necessary to meet condition a/h >=0.07, and a is the diameter of particle, and h is
The minimum dimension of microchannel, the height of microchannel is usually minimum dimension in inertia micro flow chip.It, can if particle size is too small
Particle can be caused inertia can not to focus in the microchannel of finite length.Also some researches show that, a/h < 0.5 is also restrictive condition,
The excessive channel that may result in of particle size is easy blocking.It therefore, be according to the size of separating particles come really when designing preparation
Determine the size of microchannel.
The present invention provides a kind of side that Peripheral Circulation tumour cell is enriched with from peripheral blood cascade or tandem superposition sorting
Method specifically includes: sample solution to be enriched with is from 1 sample introduction of cell entry of first single-screw shape microchannel, and sheath fluid is from first
5 sample introduction of sheath fluid entrance of the sheath fluid entrance 2 of a single-screw shape microchannel and second single-screw shape microchannel, in certain fluid
Under flow conditions, the cell outlet 8 second single-screw shape microchannel collects the fluid for being rich in target cell, and non-target cell is then
It is collected from the waste liquid outlet 7 of second single-screw shape microchannel.
The working principle of Double-spiral microchannel of the present invention is as follows: the entrance first single-screw shape microchannel
At A, blood sample enters microchannel from cell entry 1, and sheath fluid enters microchannel from sheath fluid entrance 2, due to the stream of sheath fluid entrance 2
Body flow velocity is higher than the fluid flow rate of cell entry 1, and all cells of cell entry 1 are extruded into first single-screw shape microchannel
Lateral wall, and thus enter subsequent microchannel, therefore, it can be stated that, at 0 Dien period, the start bit of all cells
Set be first single-screw shape microchannel lateral wall.Under the collective effect of inertia lift and Dien drag force, target cell due to
Its size is larger, and at 0.5 Dien period, the inner sidewall first single-screw shape microchannel, which is formed, focuses a fluid stream, and by
In the effect of its inertia lift and Dien drag force, target cell protects always in the subsequent microchannel in first single-screw shape microchannel
Hold the focused flow in its inner sidewall；Non-target cell is because its size is smaller, the mainly effect by Dien drag force, with Dien vortex
Lateral wall and inner sidewall back and forth lateral transfer of the form first single-screw shape microchannel, also, usually 0.5 Dien week
Positioned at the inner sidewall of first single-screw shape microchannel when the phase, and its initial position is back at 1 complete Dien period,
The lateral wall of i.e. first single-screw shape microchannel.Since the length of first single-screw shape microchannel is deposited with the above-mentioned Dien period
In certain proportion, in its exit, target cell is flowed out simultaneously from the cell outlet 4 for being located at first single-screw shape microchannel inner sidewall
It is directly entered the cell entry of second single-screw shape microchannel, and non-target cell fluid is then micro- from first single-screw shape is located at
The waste liquid outlet 5 of channel lateral wall is flowed out and is collected through pressure balance microchannel 3；It is thin due to first single-screw shape microchannel
Born of the same parents outlet 4 be second single-screw shape microchannel cell entry, therefore, in second single-screw shape microchannel, according to
The identical working principle of first single-screw shape microchannel, the target cell that the sorting of first single-screw shape microchannel is enriched to is the
Two single-screw shape microchannels are enriched with by the 2nd sorting or tandem type sorting is enriched with or tandem sorting is enriched with or superposing type
Sorting enrichment.It can thus be stated that the Double-spiral microchannel through the invention, the target cell in blood sample is cascaded
Or tandem carried out the sorting enrichment of 2 superposing types, simultaneously as most of non-target cell is micro- by first spiral shape
Channel removal, into total number of cells well below second single spiral shell on the whole of the cell suspension of second single-screw shape microchannel
The total number of cells of the cell suspension of shape microchannel are revolved, to sort concentration effect meeting due to cell because the interference of its density is few
It is more preferable than single spiral shape microchannel is used alone, it can finally obtain the target cell of high-purity.Note that the present embodiment with
Lower part is actually in Double-spiral channel more of the present invention and traditional single-screw shape microchannel (as shown in figure 11)
Sorting concentration effect.
In specific experiment, the width l of spiral shape microchannel is 600 μm, radius of curvature 1.15cm, microchannel height h
It is 130 μm.Traditional single-screw shape microchannel (as shown in figure 11) is in spiral planar inside configuration design entry, planar junction
Structure build up outside outlet, wherein the width of cell entry and cell outlet is 0.15mm, is located at the outer of spiral shape microchannel
The width of side wall and inner sidewall, sheath fluid entrance and waste liquid outlet is 0.45mm, be located at spiral shape microchannel inner sidewall and
Lateral wall, and cell entry and waste liquid entrance are respectively positioned on the inside of spiral shape microchannel planar geometry, cell outlet and sheath
Liquid exports the outside for being respectively positioned on spiral shape microchannel planar geometry.The cell of Double-spiral microchannel of the present invention enters
Mouthful 1 and the width of cell outlet 4 be 0.15mm, the width of sheath fluid entrance 2 and waste liquid outlet 6 is 0.45mm；Each single-screw shape
There are cell entry 1, sheath fluid entrance 5, cell outlet 4 and waste liquid outlet 6 in microchannel, wherein cell entry 1 and sheath fluid entrance 2
The entrance of each single-screw shape microchannel is pooled, cell outlet 4 and waste liquid outlet 6 pool each single-screw shape microchannel
Export B；Cell entry 1 is located at the exterior sidewall portions of set single-screw shape microchannel entrance, and sheath fluid entrance 2 is located at set single-screw
The inner sidewall part of shape microchannel entrance；Cell outlet 4 is located at the inner sidewall part of set single-screw shape microchannel outlet, waste liquid
Outlet 6 is located at the exterior sidewall portions of set single-screw shape microchannel outlet；The cell outlet 4 of first single-screw shape microchannel with
The cell entry of second single-screw shape microchannel is tightly connected into first classification cycle point；It is micro- logical by first single-screw shape
The sheath fluid entrance 5 of 6, second single-screw shape microchannels of waste liquid outlet in road and first classification cycle point are constituted the first minute
At grade circulation；It is micro- that the waste liquid outlet 6 of first single-screw shape microchannel at first classification cycle is equipped with pressure balance
Channel 3.In view of two factors of variation different in flow rate and passage length are final to different size cells generated lateral
It is displaced the difference of focal position, final optimization pass determines optimal parameter.
The production of the spiral shape micro-channel chip of two kinds of specifications is mainly by basic micrometer-nanometer processing technology, using SU8 light
Photoresist makes mold in the soft lithography of the enterprising rower standard of silicon wafer, to carrying out at punching at its corresponding entrance after PDMS demoulding
Reason, and be bonded on corresponding slide, it in order to increase the fastness of bonding, places in 70 DEG C of ovens, toasts 30min.
Spiral shape micro-channel chip entrance after bonding uses the connection type of magnetic fixture device and accurate guide pin
It is closely connect with disengaging micro-pipe, and by syringe pump and flow velocity control system to reach sample introduction stabilization and can supervise in real time
The purpose of velocity measurement variation.Before the test of its separating effect of spiral shape micro-channel chip, first using sheath flow liquid in 1 atmospheric pressure
High pressure under its leakproofness is detected, as a result without leakage situation, illustrate at chip bonding and chip entrance connection compared with
Securely, by microscopically observation chip integrality and removal of bubbles situation, after bubble excludes completely.Utilize standard polyphenyl second
Alkene colour microballon optimizes the optimal parameter of spiral shape microchannel sorting enrichment cycles tumour cell.It is micro- using 3~6 μm
Ball, 7~10 μm of microballoons, 15~18 μm of microballoons are mixed simulation red blood cell, leucocyte, circulating tumor cell respectively, using phosphorus
Phthalate buffer is diluted to 1.0*10 to 3~6 μm, 7~10 μm, 15~18 μm of microballoon respectively7Particle/milliliter, 1.0*106It is micro-
Grain/milliliter, 1.0*102A particle/milliliter, and mixed according to isometric ratio.The syringe pump and dress of mixing microballoon will be loaded
The syringe pump for carrying phosphate buffer sheath flow liquid is connected on the outside of the spiral shape chip after excluding bubble by accurate guide pin
It records and collects of the present invention respectively by flow rate monitoring system and pressure pump control adjustment flow velocity with inner sidewall entrance
Double-spiral microchannel (as shown in Figure 1), traditional single-screw shape microchannel (as shown in figure 11) under different in flow rate (i.e.:
150 μ l/min, 300 μ l/min, 450 μ l/min, 600 μ l/min, 750 μ l/min, 900 μ l/min) microballoon point in each outlet
Select enrichment condition.The result sample situation that microballoon separates at cell outlet is shown in Fig. 8, captures list because ordinary camera can not understand
The motion state of a rare cell or particle, thus in order to clearly intuitively statement rare cell or particle motion profile, with compared with
The dot of minor diameter represents 3~6 μm and 7~10 μm of rare cell or particle, represents 15~18 μm with larger-diameter dot
Rare cell or particle motion profile.Result is observed in the microscopic counting that microballoon separates at cell outlet 8 and waste liquid outlet 7
Sample situation is shown in Fig. 9.Fig. 9-a is the displaing micro picture at traditional single-screw shape microchannel cell outlet, and three kinds of microballoons are more,
Prompt the sorting concentration effect of larger particle poor, purity is low；Fig. 9-b is Double-spiral microchannel of the present invention cell outlet
The displaing micro picture at place is almost larger-diameter microballoon, prompts the sorting concentration effect of larger particle very good, purity pole
It is high；Fig. 9-c is the displaing micro picture at Double-spiral microchannel of the present invention waste liquid outlet 7, it is seen that is almost small diameter
Microballoon, almost lose larger-diameter microballoon, larger particle prompted to be enriched to cell outlet, have good recycling
Rate.Further analysis is the results show that two kinds of spiral shape microchannels all have optimal when flow velocity is in 600~750 μ l/min
Sort concentration effect；When the flow velocity of fluid is less than 600 μ l/min, portion size larger particles can not be focused on close to inner sidewall
Cell outlet；When the flow velocity of fluid is greater than 750 μ l/min, the cell purity decline of larger particle；Associated exemplary result is such as
Shown in Fig. 8.
As shown in table 1, further comparative study the result shows that, no matter under which kind of flow conditions, Double-spiral is micro- logical
It is micro- logical that the purity and repeatability of the large-size particle (that is: 15~18 μm) of road sorting enrichment are far superior to traditional single-screw shape
Road, the two have statistical significant difference.Itself main reason is that, it is single by its own in traditional single-screw shape microchannel
The limitation of spiral shape microchannel often makes the lesser particle of size that lateral displacement occur, thus the big ruler for being enriched to sorting
The repeatability of very little particle (that is: 15~18 μm) purity and experimental result is affected.But Double-spiral of the present invention is micro-
Channel because can second level sorting enrichment, thus substantially increase including purity and repeatability including detection efficiency (table 1), the two
With statistically-significant difference.Further result of practical application shows under similar experiment condition, when 8 milliliters of blood of processing
When liquid sample, traditional single-screw shape microchannel is 40 minutes time-consuming, and Double-spiral microchannel of the present invention is only 10 points time-consuming
Clock, and the repeatability of the purity of Double-spiral microchannel circulating tumor cell obtained of the present invention and experimental result is higher
(close with result described in table 1), the two have statistically-significant difference.The studies above result prompt, relative to traditional single spiral shell
Shape microchannel is revolved, the clinical practice detection that Double-spiral microchannel of the present invention was more able to satisfy and met circulating tumor cell needs
Ask and subsequent detection analysis harsh conditions (such as circulating tumor cell individual cell level genetic test).
The particle purity that the lower record different in flow rate of table one is collected into
| Flow velocity | Double helix | Single-screw |
| 150μl/min | 54.56±1.12 | 34.58±5.28 |
| 300μl/min | 76.24±2.35 | 42.56±5.81 |
| 450μl/min | 86.10±2.84 | 62.57±6.12 |
| 600μl/min | 94.12±2.08 | 67.60±7.27 |
| 750μl/min | 99.08±0.43 | 70.36±8.76 |
| 900μl/min | 97.02±2.47 | 54.56±5.21 |
Doubling dilution is distinguished to 10 using HCC827, MCF-7, SW480 tumor cell line manually cultivated2A cell/ml,
It is mixed into according to the volume ratio of 1:5 using erythrocyte cracked liquid treated normal plasma cell, using Double-spiral of the present invention
Microchannel carries out sorting enrichment, and testing result is shown in Table two using the count results of immunofluorescence technique stain smear.As the result is shown
Double-spiral microchannel of the present invention is at least up to 98% or more, and purity and above-mentioned pair to the rate of recovery of kinds of tumor cells
Spiral shape microchannel is similar, is each about 99%.
Two variety classes tumour cell of table using Double-spiral microchannel be enriched with after the rate of recovery
Compared with traditional single-screw shape microchannel, the time of Double-spiral microchannel detection of the present invention foreshortens to biography
The 1/4 of the single-screw shape microchannel of system, and sorting be enriched to the purity of rare cell or particle, the repeatability of experimental result and
The methods of rate of recovery learns parameter and is far superior to traditional single-screw shape microchannel, illustrates micro- logical relative to traditional single-screw shape
Road, Double-spiral microchannel of the present invention can preferably meet the actual demand of clinical application.
Embodiment two
The present embodiment in the present embodiment using impressed pressure pump what is different from the first embodiment is that being realized and being pressed in embodiment one
The approximate effect in dynamic balance microchannel, even if the liquid flow direction and first of the cell entry of second single-screw shape microchannel
The liquid flow direction of the cell entry 1 of a single-screw shape microchannel is consistent, and 0.7*v2≤v1≤1.3*v2；Make second list
The liquid of the liquid flow direction of the cell outlet 8 of spiral shape microchannel and the cell outlet 4 of first single-screw shape microchannel
Flow direction is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Make the waste liquid outlet 7 of second single-screw shape microchannel
Liquid flow direction and first single-screw shape microchannel waste liquid outlet 8 liquid flow direction it is consistent, and 0.7*v6≤
v5≤1.3*v6.The pressure pump is arranged at the waste liquid outlet 6 of first single-screw shape microchannel.
Embodiment three
The present embodiment is unlike the embodiments above, and in the present embodiment, whole spiral shape microchannel is by 4 single-screws
Shape microchannel composition.In fact, binary channels spiral flow channel of the present invention can further evolve into three spiral shape microchannels, four
Spiral shape microchannel, or more grade spiral shape microchannel.Included single-screw shape is micro- logical in multiple coil shape microchannel
The quantity in road is at least 2 or more, also, the center of circle of the multiple single-screw shape microchannel is located at same point, (M-1) a list
The cell outlet of spiral shape microchannel is the cell entry of m-th single-screw shape microchannel, and (M-1) a single-screw shape is micro- logical
The waste liquid outlet in road has been directly connected to pressure equalisation passage.The effect of the pressure equalisation passage is to make (M-1) a single-screw
The flow velocity of the cell entry of the cell outlet or m-th single-screw shape microchannel of shape microchannel and direction and (M-1) a spiral
The flow velocity of the cell outlet of the cell entry and m-th single-screw shape microchannel of shape microchannel and direction keep identical or pole phase
Closely, meanwhile, make m-th single-screw shape microchannel waste liquid outlet flow velocity and direction and a spiral shape microchannel (M-1) it is useless
The flow velocity of liquid outlet and direction keep identical or extremely close, to keep each spiral shape in multiple coil shape microchannel micro-
The hydrodynamics characteristic that target cell is given in channel keeps same or similar.
The present embodiment is by taking four spiral shape microchannels as an example, as shown in figure 3, the cell outlet of first single-screw shape microchannel
4 be the cell entry of second single-screw shape microchannel, and the cell outlet of second single-screw shape microchannel is the single spiral shell of third
The cell entry of shape microchannel is revolved, the cell outlet of third single-screw shape microchannel is the thin of the 4th single-screw shape microchannel
Born of the same parents' entrance, also, the waste liquid outlet of first, second and third single-screw shape microchannel is respectively provided with corresponding pressure
Balance channel 3.
Sheath fluid of the sheath fluid from 2, second single-screw shape microchannels of sheath fluid entrance of first single-screw shape microchannel
The sheath fluid entrance 17 of entrance 5, the sheath fluid entrance 15 of third single-screw shape microchannel and the 4th single-screw shape microchannel is led
Enter；Waste liquid outlet 7 of the waste liquid from 6, second single-screw shape microchannels of waste liquid outlet of first single-screw shape microchannel, third
The waste liquid outlet 16 of a single-screw shape microchannel and the waste liquid outlet 19 of the 4th single-screw shape microchannel export；Finally
Rare cell or particle after collecting sorting enrichment at the cell outlet 18 of four single-screw shape microchannels.
Example IV
The present embodiment is realized and pressure in embodiment in the present embodiment using impressed pressure pump unlike embodiment three
Balance the approximate effect in microchannel.
Embodiment five
In the present embodiment, two spiral shape microchannels are connected using concatenated mode, the of first spiral shape microchannel
The cell of first single-screw shape microchannel of the cell outlet 8 and second spiral shape microchannel of two single-screw shape microchannels
Entrance 14 is connected, and forms the place that is installed in series of two spiral shape microchannels, and is installed in series place in each spiral shape microchannel
Position on pressure equalisation passage is set.
Sheath fluid entrance 2, first from first single-screw shape microchannel of first spiral shape microchannel of the sheath fluid
The single spiral shell of first of 5, second spiral shape microchannels of sheath fluid entrance of second single-screw shape microchannel of spiral shape microchannel
Revolve the sheath fluid entrance of the sheath fluid entrance 9 of shape microchannel and second single-screw shape microchannel of second spiral shape microchannel
11 import；6, first spiral shapes of waste liquid outlet of waste liquid from first single-screw shape microchannel of first spiral shape microchannel
First single-screw shape of 7, second spiral shape microchannels of waste liquid outlet of second single-screw shape microchannel of microchannel is micro-
The waste liquid outlet 13 of second single-screw shape microchannel of the waste liquid outlet 10 in channel and second spiral shape microchannel is led
Out.It is collected from the cell outlet 12 of second single-screw shape microchannel of second spiral shape microchannel rare after sorting is enriched with
Cell or particle.
More N grades of spiral shape microchannel can also be arranged in series using the structure type of the present embodiment, and it is existing
Some cell entries compare (example with the inner side and outer side that cell outlet is located at single spiral flow channel planar geometry
Such as: the last one attached drawing), the present invention can under conditions of not using any external micro-pipe directly cascade or tandem using multiple
N grades of spiral flow channels, not only simplify manufacture craft, also reduce the complexity of N grades of spiral flow channels, improve working efficiency and inspection
Survey the accuracy of result.
Embodiment six
The present embodiment is realized using impressed pressure pump in the present embodiment and is pressed in embodiment five unlike embodiment five
The approximate effect in dynamic balance microchannel.
Embodiment seven
In the present embodiment, two Double-spiral microchannels are connected by the way of in parallel.Sample solution is from two double helixs
The cell entry 1 of first single-screw shape microchannel of shape microchannel enters, and rare cell or particle are micro- from two Double-spirals
The cell outlet 12 of second single-screw shape microchannel in channel flows out.Participate in the center of circle projection of Double-spiral microchannel in parallel
It is overlapped.2,3 or more samples to be detected can be handled simultaneously on same micro-fluidic chip, to improve detection efficiency
While be effectively reduced processing time of testing cost and sample.
Embodiment eight
The present embodiment what is different from the first embodiment is that in this programme all entrances (include: cell entry and outlet,
Sheath fluid entrance and waste liquid outlet) it is arranged on the outside of the geometry of one-piece auger shape microchannel, waste liquid outlet is flat by pressure
Weighing apparatus fluid channel is circulated on the outside of the geometry of one-piece auger shape microchannel along the extending direction of other fluid channels, by pressure
The adjustment to cell outlet content flow velocity and flow direction is realized in the adjustment of the width, depth and length of balance fluid channel.
In the specific implementation process, by part entrance, (such as: second level sheath fluid entrance 5 shown in FIG. 1 and the first order are useless
Liquid waste liquid 6) it is arranged in that the structure on the inside of spiral shape microchannel is designed as the molding of chip and manufacture brings very big inconvenience, in core
The central hole of piece simultaneously exports or imports fluid and/or the operation of cell is also complex, leads to the system of relevant chip fixture
Make difficulty height, it is difficult to observe first order outlet and the fluid of second level entrance intersection and/or the flow regime of cell.Therefore exist
All entrances (including: cell entry and outlet, sheath fluid entrance and waste liquid outlet) are arranged at whole spiral shell in the present embodiment
Revolve the outside of shape microchannel.
Enrichment cycles tumour cell is sorted based on inertia migration force, it is imitative using the more physical quantitys of COMSOL Multiphisics
True software is further improved chip structure and successfully constructs standardized on the basis of constructing standardization single-screw chip
Double-spiral microchannel chip.Its crucial purpose of the standardization double helix chip of design construction is to be focused by inertia so that size
Biggish target cell (CTC) along channel inner wall be distributed focus, the lesser haemocyte of size kept away on the outside of helical duct with
Sheath flow liquid separation.Double helix chip is compared compared with single-screw chip, can more fast and efficiently be captured by cascade mode
CTC in blood sample meets clinical practice requirement, reaches to the monitoring of tumor patient dynamic realtime, selection therapeutic regimen
Purpose, while can also further to the related mechanisms such as metastases carry out further investigated research.
The position that its inertia of different cells focuses is influenced by the three-dimensional dimension of spiral shape fluid channel.Pass through COMSOL
The simulation calculation of Multiphisics software, it is determined that CTC (>=15um) can be made to separate effect with haemocyte (3~15um)
The optimal spiral shape microchannel chip three-dimensional dimension of fruit, to force sample can be from the cell of first single-screw shape microchannel
Entrance can smoothly enter into spiral shape microchannel completely and start its lateral displacement in runner.In view of variation different in flow rate
And two factors of length generated lateral displacement focal position final to different size cells (CTC, RBC, WBC) is not
Together, final optimization pass determines the optimal parameter of the cell outlet of second single-screw shape microchannel.Binary channels spiral chip is in single-pass
Chip inner flow passage length is increased on road spiral chip basis, avoid big flow velocity it is high-throughput under because runner it is too short caused by size
The excessive problem for causing cell purity low of smaller particle lateral displacement.
Using COMSOL Multiphysics V5.2a software, pressure equalisation passage 3 of the simulation analysis in different in width
Under the conditions of, the first order entrance of double helix fluid channel (includes: cell entry 1, sheath fluid entrance 2, waste liquid outlet 6, cell outlet
4) with the volume flow feature of second level entrance (including: cell entry 4, sheath fluid entrance 5, waste liquid outlet 7, cell outlet 8)
And related ratio.Simulated conditions are that the primary condition of cell entry is 150ul/min, and the primary condition of sheath fluid entrance is 750ul/
min.Analog result shows only when the width of pressure equalisation passage 3 is in 350um, the flow velocity of first order cell outlet 4 just with
The flow velocity of first order cell entry 1 is suitable, and with this condition, flow velocity is 144.18ul/min (table three).Should the result shows that, pressure
The width in dynamic balance channel 3 has important adjustment effect to the flow velocity of first order cell outlet 4, also, only appropriate at it
Width conditions under, just can ensure that the flow velocity of first order cell outlet 4 is suitable with the flow velocity of first order cell entry 1.Meanwhile into
The width of one step intensified pressure balance channel 3 will lead to first order cell outlet 4 in opposite direction flowing, illustrate that pressure balance is logical
The balance pressure that road 3 provides is too small, leads to the reflux of fluid.It is other the result shows that, when the width of pressure equalisation passage 3 maintains
When 250~400um, the volume flow ratio of second level waste liquid outlet 7 and cell outlet 8 maintains more constant range, tool
Body the results are shown in Table four.Further sunykatuib analysis the result shows that, the depth and length of pressure equalisation passage 3 can equally influence first
The flow velocity of grade cell outlet 4.
Table three is under conditions of pressure equalisation passage 3 of different in width, the volume flow feature of entrances at different levels
Table four is under conditions of pressure equalisation passage 3 of different in width, the volume flow feature of outlets at different levels
The production of spiral shape microchannel chip is mainly by basic micrometer-nanometer processing technology, using SU8 photoresist in silicon wafer
The soft lithography of enterprising rower standard makes mold, to carrying out punching processing at its corresponding entrance after PDMS demoulding, and is bonded
It on corresponding slide (45mm X45mm), in order to increase the fastness of bonding, places in 70 DEG C of ovens, toasts 30min.
Spiral shape microchannel chip entrance after bonding using Magnetic probe (Corsolution) device and
Fluidic Connect (Micronit) fixture connection type is closely connect with sample feeding pipe, and is infused by LongerPump
Pump and MCFS pressure flow velocity control system are penetrated to achieve the purpose that sample introduction is stablized and can real-time monitoring change in flow.To spiral
Before the test of its separating effect of shape microchannel chip, its leakproofness is detected under 1bar high pressure using sheath flow liquid first,
As a result without leakage situation, illustrate at chip bonding and the connection of chip entrance is stronger, it is complete by microscopically observation chip
Property and removal of bubbles situation, after bubble excludes completely.Spiral shape fluid channel is sorted using standard polystyren colour microballon
The optimal parameter of enrichment CTC optimizes.
Using 3~6um microballoon, 7~10um microballoon, 15~18um microballoon mixed respectively simulate red blood cell, leucocyte,
CTC, wherein 1.0 is diluted to respectively to 3~6 μm, 7~10 μm, 15~18 μm of microballoon using PBS buffer solution ×
109particles/ml、1.0×107particles/ml、1.0×103Particles/ml is simultaneously mixed according to isometric ratio
It closes.The syringe pump for loading mixing microballoon and the syringe pump for loading PBS sheath flow liquid are connected to exclusion gas by chip fixture
The cell entry 1 and sheath fluid entrance 2 of spiral shape chip after bubble pass through flow rate monitoring system and pressure pump control adjustment stream
Speed and pressure, record respectively and collect single, double screw type chip (single-screw cake core and the present embodiment in embodiment one) and exist
(200 μ l/min, 350 μ l/min, 500 μ l/min, 650 μ l/min, 800 μ l/min, 950 μ l/min), chip under different in flow rate
The coloured microballoon separation situation in exit is shown in Figure 23, and observes result by microscopic counting and see Figure 24.As the result is shown when flow velocity exists
Separating effect is best when 650~800 μ l/min.
Flow velocity can not be focused on less than 650 μ l/min portion size larger particles close to cell outlet, when flow velocity is greater than 800
When μ l/min, CTC sorting enrichment purity will be affected as shown in figure 23, and dot is how many and size does not reflect that particle is actual
Quantity number and size.
Using the studies above method, binary channels screw type chip is tested, result statistics such as table five reflects double
Its sorting enrichment purity of channel spiral cake core is much higher than single channel screw type chip.Simultaneously because single-screw chip stream road length
The limitation of degree issues the lesser particle of size in the effect of larger inertia migration force when flow velocity is greater than 800 μ l/min
The CTC cell purity that raw lateral displacement causes sorting to be enriched to is affected, however double helix chip increases flow channel length, into
The separation of the row second level is to greatly improve detection flux.The result shows that in the same time, single channel screw type chip processing energy
It is enough that 8ml blood sample is handled in 20min, and binary channels screw type chip only needs 10min, and its high-purity, high-throughput richness
Collection result, which more meets, meets the harsh conditions that CTC is applied to the analysis of clinical and subsequent detection.
The lower record different in flow rate of table five is collected into the purity of particle
Doubling dilution is distinguished to 10 using HCC827, MCF-7, SW480 cell strain manually cultivated2Cells/ml, according to
The volume ratio of 1:5 is mixed into using erythrocyte cracked liquid treated normal plasma cell, is examined using the chip in the present embodiment
It surveys, testing result is shown in Table six using the counting of immunofluorescence technique stain smear.Double helix microchannel chip is to a variety of as the result is shown
It is swollen by being mixed into that the rate of recovery of tumour cell show the SW480 immunofluorescence dyeing for sorting and being enriched to up to 90% or more, Figure 25
Its visible with the cell results captured of oncocyte has good linear correlation, while double helix chip capture is to kinds of tumors
Cell capture efficiency is shown in Figure 26 up to 90%.
Six double helix chip kinds of tumor cells of table captures the rate of recovery
In addition, being detected respectively to 20 colorectal cancer patients samples, wherein utilizing standardization single-screw and double helix
Chip all detects wherein 18 patient's CTC cells, but compares compared with single-screw chip, and double helical form chip detection time shortens one
Half, close to meeting the needs of clinical high-throughput.Meanwhile the double helix channel of our designs, it is designed by standardization inlet and outlet,
The chip fixture being largely commercialized can be cooperated to be precisely accomplished whole processes that entire blood sample sample introduction goes out sample, make it
From clinical a large amount of samples, low cost, quickly detection CTC is further, can 10min handle 8ml blood sample its to standardization
The sorting enrichment purity of microballoon is up to 98% or more, and in theory, sorting is enriched with CTC purity equally up to 98% or more, but
Because otherness is larger between tumor patient (WBC significantly increases after such as chemotherapy), therefore its purity is often a variable but compares
Single-screw cake core, double helical form die grading enrichment CTC cell purity are much higher than single-screw chip, cell purity pair
The biology manipulation analysis such as subsequent science of heredity of CTC cell is most important.
Embodiment nine
The present embodiment is more similar in itself to embodiment five, but all entrances in this programme (include: that cell enters
Mouth and outlet, sheath fluid entrance and waste liquid outlet) it is arranged on the outside of the geometry of one-piece auger shape microchannel, it is easier to it observes
With analysis as a result, simultaneously, standardization fixture is also easier to, to be advantageously implemented mass handling and operation.
Embodiment ten
The present embodiment is more similar in itself to embodiment three, but the design is by the thin of first single-screw shape microchannel
Born of the same parents' entrance, the sheath fluid entrance of first single-screw shape microchannel, second single-screw shape microchannel sheath fluid entrance, first list
The position being closer to is arranged in the waste liquid outlet of spiral shape microchannel, and two single-screw shape microchannels are arranged with concentric
Form is arranged alternately, meanwhile, all entrances (including: cell entry and outlet, sheath fluid entrance and waste liquid outlet) are arranged at
On the outside of the geometry of one-piece auger shape microchannel, overall structure is more exquisite, is easy to use.
Embodiment 11
The structure that the present embodiment is attached by the way of in parallel by the Double-spiral microchannel in two embodiments eight
Schematic diagram, using the form of above-mentioned parallel connection can make the same sample within the unit time simultaneously two Double-spiral microchannels into
Row processing, in other words, can realize within the time of primary screening and handle while more fluid, can effectively increase screening effect
Rate.Relative to embodiment eight, the detection time of clinical sample can at least shorten 1 times.Two double helixs are only shown in the present embodiment
The parallel form of shape microchannel, can according to need progress three, four in actual use and more a plurality of Double-spiral is micro- logical
The parallel connection in road.
Embodiment 12
The present embodiment is more similar in itself to embodiment seven, but the design is by the of first Double-spiral microchannel
The cell entry of one single-screw shape microchannel, the sheath fluid entrance of first single-screw shape microchannel, second single-screw shape are micro-
The sheath fluid entrance in channel, first single-screw shape microchannel waste liquid outlet the position being closer to is set；It is double by second
The cell entry of first single-screw shape microchannel in spiral shape microchannel, the sheath fluid of first single-screw shape microchannel enter
Mouthful, the waste liquid outlet setting of the sheath fluid entrance of second single-screw shape microchannel, first single-screw shape microchannel more connecing
Close position, and two single-screw shape microchannels are arranged and are arranged alternately in concentric form, overall structure is more exquisite, is convenient for
It uses.Meanwhile all entrances (including: cell entry and outlet, sheath fluid entrance and waste liquid outlet) in this programme are respectively provided with
On the outside of the geometry of one-piece auger shape microchannel, it is easier to which observation and analysis are as a result, simultaneously, be also easier to standardization folder
Tool, to be advantageously implemented mass handling and operation.
Embodiment 13
The present embodiment is by the spiral shape microchannel (i.e. concatenated double helix microchannel) in two embodiments ten using in parallel
The structural schematic diagram that is attached of mode, the same sample can be made to exist simultaneously within the unit time using the form of above-mentioned parallel connection
Spiral shape microchannel shown in two embodiments ten is handled, and in other words, more multithread can be realized within the time of primary screening
It is handled while body, screening efficiency can be effectively increased.Relative to embodiment ten, the detection time of clinical sample can at least shorten
1 times.The present embodiment had both had to be guaranteed repeatedly to screen using multi-stage path by concatenated form, improves the advantage of cell purity,
Also there is the advantage that screening efficiency is improved by way of in parallel.Spiral shape shown in two embodiments ten is only shown in the present embodiment
The parallel form of microchannel can according to need spiral shell shown in progress three, four and more a plurality of embodiment ten in actual use
Revolve the parallel connection of shape microchannel.
Embodiment 14
Another way of realization in basic engineering-single-screw shape microchannel is shown in the present embodiment, it will be thin in this kind of form
Born of the same parents outlet and waste liquid outlet are arranged at the outside of one-piece auger shape microchannel.In the specific implementation process, by cell outlet
And waste liquid outlet is arranged at the design of the structure on the inside of the spiral shape microchannel molding and manufacture band (as shown in figure 11) for chip
Carried out very big inconvenience, chip central hole and export or import fluid and/or cell operation it is also complex, lead to phase
The manufacture difficulty of the chip fixture of pass is high, it is difficult to observe the fluid of sheath fluid and cell entry intersection and/or the flowing shape of cell
State.Therefore cell outlet and waste liquid outlet are arranged to the outside of one-piece auger shape microchannel in the present embodiment, it is easier to
Observation and analysis, and standardization fixture and easy operation.
Embodiment 15
Show that the single-screw shape microchannel in two embodiments 14 is attached using concatenated mode in the present embodiment
Structural schematic diagram, may be implemented repeatedly to screen in a flow process using concatenated form, can further improve screening
The purity of cell.The cascade that two single-screw shape microchannels are only shown in the present embodiment, in actual use can basis
Need to carry out the series connection of three, four and more a plurality of single-screw shape microchannel.
Embodiment 16
Another form of expression of embodiment 15 is shown in the present embodiment, two single-screw shape microchannels use with one heart simultaneously
Column wrap setting, tortuous folding helically coiling cyclization, may be implemented repeatedly to sieve in a flow process using concatenated form
Choosing can further improve the purity of screening cell.The cascade that two single-screw shape microchannels are only shown in the present embodiment,
It can according to need the series connection for carrying out three, four and more a plurality of single-screw shape microchannel in actual use.
Embodiment 17
Show that the single-screw shape microchannel in two embodiments 14 is attached by the way of in parallel in the present embodiment
Structural schematic diagram, using the form of above-mentioned parallel connection can make the same sample within the unit time simultaneously in two embodiments 14
Shown single-screw shape microchannel is handled, in other words, while can realizing more fluid within the time of primary screening at
Reason, can effectively increase screening efficiency.Relative to embodiment 14, the detection time of clinical sample can at least shorten 1 times.This reality
The parallel form for only showing two single-screw shape microchannels in example is applied, can according to need carry out three, four in actual use
The parallel connection of item and more a plurality of single-screw shape microchannel.
Embodiment 18
Another form of expression of embodiment 17 is shown in the present embodiment, two single-screw shape microchannels use with one heart simultaneously
Column wrap setting, tortuous folding helically coiling cyclization, can realize more multithread using form in parallel within the primary screening time
It is handled while body, screening efficiency can be effectively increased, the shape in parallel of two single-screw shape microchannels is only shown in the present embodiment
Formula can according to need the parallel connection for carrying out three, four and more a plurality of single-screw shape microchannel in actual use.
Basic engineering of the invention be single-screw shape microchannel, extending design be single-screw shape microchannel series connection,
The series, parallel of in parallel, mixed connection and more spiral shape microchannels (in description of the invention by taking Double-spiral microchannel as an example),
A variety of ways of realization such as mixed connection, the above-mentioned form of expression are the extending design of this programme, and basic principle is in the present specification
It is set forth, therefore the above-mentioned various forms of expression should all also be fallen among protection scope of the present invention.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to compared with
Good embodiment describes the invention in detail, those skilled in the art should understand that, it can be to skill of the invention
Art scheme is modified or replaced equivalently, and without departing from the objective and range of the technical program, should all be covered in the present invention
Scope of the claims in.
Claims (82)
1. a kind of spiral shape microchannel, for sorting enrichment rare cell or particle, it is characterised in that:
Including N number of single-screw shape microchannel, microchannel that single single-screw shape microchannel includes entrance, connect with inlet seal
And with channel far from entrance one end be tightly connected outlet, the entrance by cell entry and sheath fluid entrance convergence and
At the outlet is converged by waste liquid outlet and cell outlet；
The cell outlet of (M-1) a single-screw shape microchannel and the cell entry of m-th single-screw shape microchannel are tightly connected
At (M-1) a classification cycle point, by the waste liquid outlet of (M-1) a single-screw shape microchannel, m-th single-screw shape microchannel
Sheath fluid entrance and (M-1) a classification cycle point constitute at (M-1) a classification cycle；
The waste liquid outlet of (M-1) a single-screw shape microchannel at (M-1) a classification cycle is equipped with pressure balance mould
Block, the flow rate of liquid of the cell entry of m-th single-screw shape microchannel are v1, the cell of (M-1) a single-screw shape microchannel
The flow rate of liquid of entrance is v2；The flow rate of liquid of the cell outlet of m-th single-screw shape microchannel is v3, (M-1) a single spiral shell
The flow rate of liquid for revolving the cell outlet of shape microchannel is v4；The flow rate of liquid of the waste liquid outlet of m-th single-screw shape microchannel is
V5, the flow rate of liquid of the waste liquid outlet of (M-1) a single-screw shape microchannel are v6；
The pressure balance module is used to make the liquid flow direction and (M-1) of the cell entry of m-th single-screw shape microchannel
The liquid flow direction of the cell entry of a single-screw shape microchannel is consistent, and 0.7*v2≤v1≤1.3*v2；Make m-th list spiral shell
Revolve the liquid flow of the liquid flow direction of the cell outlet of shape microchannel and the cell outlet of (M-1) a single-screw shape microchannel
Dynamic direction is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Make the liquid of the waste liquid outlet of m-th single-screw shape microchannel
Body flow direction is consistent with the liquid flow direction of waste liquid outlet of (M-1) a single-screw shape microchannel, and 0.7*v6≤v5
≤1.3*v6；
N number of single-screw shape microchannel forms non-cross spiral shape microchannel；
Wherein, the cell entry of first single-screw shape microchannel is used to enter the sample liquid containing rare cell or particle；N
The cell outlet of a single-screw shape microchannel is used to collect the rare cell or particle after sorting enrichment；The sheath fluid entrance is used for
Addition has osmotic pressure, guarantees rare cell or the complete dilution of particle；1 < M≤N, M, N is positive integer.
2. spiral shape microchannel as described in claim 1, it is characterised in that: the pressure balance module is setting the
(M-1) at a classification cycle in (M-1) a single-screw shape microchannel waste liquid outlet at and with the waste liquid outlet seal connect
The pressure balance microchannel connect；It is a that the pressure balance microchannel is equipped with (N-1).
3. spiral shape microchannel as described in claim 2, it is characterised in that: the pressure balance microchannel and N number of single spiral shell
Revolve the setting of shape microchannel in the same plane.
4. spiral shape microchannel as described in claim 1, it is characterised in that: the pressure balance module is setting the
(M-1) at a classification cycle in (M-1) a single-screw shape microchannel waste liquid outlet at pressure pump.
5. spiral shape microchannel as described in claim 1, it is characterised in that: the entrance of first single-screw shape microchannel and
The outlet of n-th single-screw shape microchannel is located at the outside of spiral shape microchannel geometry.
6. spiral shape microchannel as described in claim 1, it is characterised in that: the spiral shape microchannel is by N number of single spiral shell
Rotation shape microchannel forms the ring-type being parallel to each other.
7. spiral shape microchannel as described in claim 6, it is characterised in that: single spiral shell between the spiral shape microchannel
Revolving the horizontal distance between shape microchannel and single-screw shape microchannel adjacent thereto and parallel is 0.1~1000 μm.
8. spiral shape microchannel as described in claim 1, it is characterised in that: the section of single single-screw shape microchannel is
Rectangle is trapezoidal.
9. spiral shape microchannel as described in claim 8, it is characterised in that: the section of the single-screw shape microchannel is
Rectangle, ratio w/h=1.2~10 of the width w and height h of the rectangle.
10. spiral shape microchannel as described in claim 8, it is characterised in that: the section of the single-screw shape microchannel
To be trapezoidal, the side height of the geometric center of the trapezoidal close spiral shape microchannel is micro- lower than the trapezoidal separate spiral shape logical
The side height of the geometric center in road.
11. spiral shape microchannel as described in claim 9, it is characterised in that: the width w of microchannel is 0.1-1000 μm,
Height is 0.1-800 μm.
12. spiral shape microchannel as described in claim 1, it is characterised in that: N=2, the spiral shape microchannel is by
One single-screw shape microchannel and second single-screw shape microchannel composition.
13. spiral shape microchannel as described in claim 1, it is characterised in that: the entrance of single single-screw shape microchannel
And the distance between outlet of the single-screw shape microchannel is 1.7-2.0 of non-target cell or non-target particles hydrodynamics characteristic
The Dien period, alternatively, 2.7-3.0 Dien period, alternatively, 3.7-4.0 Dien period, alternatively, 4.7-5.0 Dien week
Phase, alternatively, 5.7-6.0 Dien period, alternatively, 6.7-7.0 Dien period, alternatively, 7.7-8.0 Dien period, alternatively,
The 8.7-9.0 Dien period, alternatively, 9.7-10.0 Dien period.
14. spiral shape microchannel as described in claim 1, it is characterised in that: the channel of single-screw shape microchannel is cut
Face width maximum width is l1, and the cell entry of the single-screw shape microchannel and/or the width of cell outlet are l2,0.1*l1
≤l2≤0.4*l1；The width of the sheath fluid entrance of the single-screw shape microchannel and/or waste liquid outlet is l3,0.1*l1≤l3≤
0.9*l1。
15. spiral shape microchannel as described in claim 1, it is characterised in that: N=4, the spiral shape microchannel is by
One single-screw shape microchannel, second single-screw shape microchannel, third single-screw shape microchannel and the 4th single-screw
Shape microchannel composition.
16. spiral shape microchannel as described in claim 1, it is characterised in that: the diameter of the rare cell or particle is
A, the height of the microchannel are h, 0.07≤a/h≤0.5.
17. spiral shape microchannel as described in claim 1, it is characterised in that: the rare cell or particle are swollen for circulation
Oncocyte, circulation progenitor cells, circulation fetal cell, cell vesicle, excretion body or pathogen.
18. spiral shape microchannel as described in claim 1, it is characterised in that: further include for the spirality channel institute
Sort the detection unit that the rare cell being enriched to or particle are identified and/or counted；The detection unit is arranged in n-th
At the cell outlet of single-screw shape microchannel.
19. spiral shape microchannel as described in claim 18, it is characterised in that: the detection unit passes through particular technology hand
The Characters Identification of Duan Jinhang rare cell or particle；The particular technology means are selected from by Photobiology, electrobiology, biology
The group of acoustics, biomagnetism and any combination thereof composition.
20. spiral shape microchannel as described in claim 18, it is characterised in that: the detection unit passes through to special characteristic
Detection carry out the identification of rare cell or particle proterties, the special characteristic is selected from the Terahertz by rare cell or particle
The group that wave spectrum, Raman spectrum, impedance, differential impedance and any combination thereof form.
21. spiral shape microchannel as described in claim 2, it is characterised in that: a described pressure balance microchannel (N-1)
Non-cross spiral shape microchannel, the outlet of one or more pressure balance microchannels are formed with N number of single-screw shape microchannel
It is arranged in outside the geometric figure of spiral shape microchannel, by adjusting the length and internal diameter of a pressure balance microchannel (M-1)
To adjust the flow velocity in (M-1) a cell outlet by content.
22. the spiral shape microchannel as described in claim 21, it is characterised in that: (N-1) a described pressure balance is micro- logical
The outlet in road is arranged at outside the geometric figure of spiral shape microchannel.
23. spiral shape microchannel as described in claim 1, it is characterised in that: one or more in N number of cell outlet
Outside a geometric figure that spiral shape microchannel is set.
24. the spiral shape microchannel as described in claim 23, it is characterised in that: all cell outlets are arranged at spiral shape
Outside the geometric figure of microchannel.
25. spiral shape microchannel as described in claim 1, it is characterised in that: one or more settings in N number of entrance exist
Outside the geometric figure of spiral shape microchannel.
26. spiral shape microchannel as recited in claim 25, it is characterised in that: all cell entries are arranged at spiral shape
Outside the geometric figure of microchannel.
27. spiral shape microchannel as recited in claim 25, it is characterised in that: all sheath fluid entrances are arranged at spiral shape
Outside the geometric figure of microchannel.
28. the application method of spiral shape microchannel as described in any one of claim 1~27, which is characterized in that including with
Lower step:
S1: sample liquid is imported from the cell entry of first single-screw shape microchannel, by dilution from first single-screw
The sheath fluid entrance of the sheath fluid entrance of shape microchannel to n-th single-screw shape microchannel imports；
S2: the cell outlet in n-th single-screw shape microchannel collects the fluid rich in rare cell or particle；In first list
Waste liquid is discharged in the waste liquid outlet of the waste liquid outlet of spiral shape microchannel to n-th single-screw shape microchannel.
29. the application method of spiral shape microchannel as claimed in claim 28, which is characterized in that the sample used in step S1
This liquid is the blood sample with certain hematid specific volume, or the blood sample of a large amount of red blood cells is removed by haemolysis mode
This；Hematid specific volume in the blood sample is 0%~70%.
30. the application method of spiral shape microchannel as claimed in claim 28, which is characterized in that is used in step S1 is dilute
It is phosphate buffer, culture serum or physiological saline for protecting haemocyte integrality that liquid, which is released, with certain osmotic pressure.
31. the application method of spiral shape microchannel as claimed in claim 28, which is characterized in that it further include step S1.1,
S1.1: adjustment pressure balance module, make the cell entry of the m-th single-screw shape microchannel liquid flow direction and
The liquid flow direction of the cell entry of (M-1) a single-screw shape microchannel is consistent, and 0.7*v2≤v1≤1.3*v2；Make
The liquid flow direction of the cell outlet of M single-screw shape microchannel and the cell outlet of (M-1) a single-screw shape microchannel
Liquid flow direction it is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Make the waste liquid of m-th single-screw shape microchannel
The liquid flow direction of the waste liquid outlet of the liquid flow direction and (M-1) a single-screw shape microchannel of outlet is consistent, and
0.7*v6≤v5≤1.3*v6。
32. the structure that is installed in series of the spiral shape microchannel as described in claim 1~27, which is characterized in that use P spiral shell
The mode that rotation shape microchannel is arranged in series, the cell outlet of the n-th single-screw shape microchannel of a spiral shape microchannel (Q-1)
It is connected with first single-screw shape microchannel entrance of the Q spiral shape microchannel, forms spiral shape microchannel and be installed in series
Place, 1 < Q≤P.
33. the structure that is installed in series of the spiral shape microchannel as described in claim 32, which is characterized in that micro- logical in spiral shape
It also is provided with pressing at the waste liquid outlet of the n-th single-screw shape microchannel of a spiral shape microchannel (Q-1) at the road place of being installed in series
Dynamic balance module.
34. the structure that is installed in series of the spiral shape microchannel as described in claim 33, which is characterized in that the pressure balance
Module be installed in series with spiral shape microchannel place a spiral shape microchannel (Q-1) n-th single-screw shape microchannel
The pressure balance microchannel that waste liquid outlet is tightly connected.
35. the structure that is installed in series of the spiral shape microchannel as described in claim 34, which is characterized in that the pressure is flat
Weighing apparatus microchannel and matched spiral shape microchannel form non-cross spiral shape microchannel, and one or more pressure are flat
Weighing apparatus microchannel outlet be arranged in outside the geometric figure of spiral shape microchannel, by adjusting pressure balance microchannel length and
Internal diameter adjusts the flow velocity in matched cell outlet by content.
36. the structure that is installed in series of the spiral shape microchannel as described in claim 35, which is characterized in that all pressure balances are micro-
The outlet in channel is arranged at outside the geometric figure of spiral shape microchannel.
37. the structure that is installed in series of the spiral shape microchannel as described in claim 32, it is characterised in that: the cell outlet
In one or more be arranged in outside the geometric figure of spiral shape microchannel.
38. the structure that is installed in series of the spiral shape microchannel as described in claim 37, it is characterised in that: all cell outlets
It is arranged at outside the geometric figure of spiral shape microchannel.
39. the structure that is installed in series of the spiral shape microchannel as described in claim 32, it is characterised in that: in the entrance
One or more is arranged in outside the geometric figure of spiral shape microchannel.
40. the structure that is installed in series of the spiral shape microchannel as described in claim 39, it is characterised in that: all cell entries
It is arranged at outside the geometric figure of spiral shape microchannel.
41. the structure that is installed in series of the spiral shape microchannel as described in claim 39, it is characterised in that: all sheath fluid entrances
It is arranged at outside the geometric figure of spiral shape microchannel.
42. the structure that is installed in series of the spiral shape microchannel as described in claim 33, which is characterized in that the pressure balance
Module be arranged in spiral shape microchannel be installed in series place a spiral shape microchannel (Q-1) n-th single-screw shape it is micro- logical
Pressure pump at the waste liquid outlet in road.
43. the structure that is installed in series of the spiral shape microchannel as described in claim 32, which is characterized in that N=2, the spiral shell
Rotation shape microchannel is Double-spiral microchannel.
44. the structure that is installed in series of the spiral shape microchannel as described in claim 32, which is characterized in that P=2, using 2
The mode that spiral shape microchannel is arranged in series, the cell outlet of the n-th single-screw shape microchannel of first spiral shape microchannel
It is connected with first single-screw shape microchannel entrance of second spiral shape microchannel.
45. spiral shape microchannel is installed in parallel structure as described in any one of claim 1~27, which is characterized in that adopt
The mode being arranged in parallel with X spiral shape microchannel, first single-screw shape of sample solution from X spiral shape microchannel are micro- logical
The cell entry in road enters, and rare cell or particle go out from the cell of the n-th single-screw shape microchannel of X spiral shape microchannel
Mouth outflow；X > 1.
46. spiral shape microchannel as described in claim 45 is installed in parallel structure, which is characterized in that by X spiral shape
The cell entry of first single-screw shape microchannel of microchannel is interconnected.
47. spiral shape microchannel as described in claim 45 is installed in parallel structure, which is characterized in that by X spiral shape
The cell outlet of the n-th single-screw shape microchannel of microchannel is interconnected.
48. spiral shape microchannel as described in claim 45 is installed in parallel structure, which is characterized in that the X spiral shell
Revolve the setting of shape microchannel in the same plane.
49. spiral shape microchannel as described in claim 48 is installed in parallel structure, which is characterized in that the X spiral shell
Rotation shape microchannel is arranged concentrically in the same plane.
50. spiral shape microchannel as described in claim 45 is installed in parallel structure, which is characterized in that the X spiral shell
It revolves shape microchannel with one heart and is arranged along the direction perpendicular to single spiral shape microchannel place plane.
51. spiral shape microchannel as described in claim 46 is installed in parallel structure, which is characterized in that X=2, using two
The mode that a spiral shape microchannel is arranged in parallel, first single-screw shape microchannel of the sample solution from two spiral shape microchannels
Cell entry enter, rare cell or particle go out from the cell of the n-th single-screw shape microchannel of two spiral shape microchannels
Mouth outflow.
52. spiral shape microchannel as described in claim 46 is installed in parallel structure, which is characterized in that N=2, using X
The mode that Double-spiral microchannel is arranged in parallel, first single-screw shape of sample solution from X Double-spiral microchannel are micro- logical
The cell entry in road enters, and rare cell or particle are thin from second single-screw shape microchannel of X Double-spiral microchannel
Born of the same parents export outflow.
53. spiral shape microchannel as described in claim 45 is installed in parallel structure, which is characterized in that micro- logical in spiral shape
Pressure balance module is equipped at the waste liquid outlet of the n-th single-screw shape microchannel of X spiral shape microchannel in road.
54. spiral shape microchannel as described in claim 53 is installed in parallel structure, which is characterized in that the pressure balance
Module is pressure pump.
55. spiral shape microchannel as described in claim 53 is installed in parallel structure, which is characterized in that the pressure balance
Module is pressure balance microchannel.
56. spiral shape microchannel as described in claim 55 is installed in parallel structure, which is characterized in that the pressure is flat
Weighing apparatus microchannel and matched spiral shape microchannel form non-cross spiral shape microchannel, and one or more pressure are flat
Weighing apparatus microchannel outlet be arranged in outside the geometric figure of spiral shape microchannel, by adjusting pressure balance microchannel length and
Internal diameter adjusts the flow velocity in matched cell outlet by content.
57. spiral shape microchannel road is installed in parallel structure as described in claim 56, which is characterized in that all pressure balances
The outlet of microchannel is arranged at outside the geometric figure of spiral shape microchannel.
58. spiral shape microchannel road as described in claim 45 is installed in parallel structure, it is characterised in that: the cell goes out
One or more in mouthful is arranged in outside the geometric figure of spiral shape microchannel.
59. spiral shape microchannel road as described in claim 58 is installed in parallel structure, it is characterised in that: all cells go out
Mouth is arranged at outside the geometric figure of spiral shape microchannel.
60. spiral shape microchannel road as described in claim 45 is installed in parallel structure, it is characterised in that: in the entrance
One or more be arranged in outside the geometric figure of spiral shape microchannel.
61. spiral shape microchannel road as described in claim 60 is installed in parallel structure, it is characterised in that: all cells enter
Mouth is arranged at outside the geometric figure of spiral shape microchannel.
62. spiral shape microchannel road as described in claim 60 is installed in parallel structure, it is characterised in that: all sheath fluids enter
Mouth is arranged at outside the geometric figure of spiral shape microchannel.
63. a kind of combination connecting installation structure in spiral shape microchannel road, it is characterised in that: described including K microchannel monomer
Microchannel monomer is spiral shape microchannel described in any one of claim 1~27；The microchannel monomer, which uses, connects, simultaneously
Connection, repeatedly series connection, type of attachment repeatedly in parallel or combining in series and parallel are combined with each other, and wherein K is positive integer and K >=2；Series connection
Type of attachment be by first cell of the n-th cell outlet of (K-1) a microchannel monomer and k-th microchannel monomer
Inlet seal connection, forms tandem spiral shape microchannel；Type of attachment in parallel is by the of (K-1) a microchannel monomer
The sealed connection convergence of first cell entry of one cell entry and k-th microchannel monomer is combined into unified cell and enters
Mouthful；By first sheath fluid inlet seal of first sheath fluid entrance of (K-1) a microchannel monomer and k-th microchannel monomer
Connection convergence is combined into unified sheath fluid entrance, forms parallel spiral shape microchannel.
64. the combination connecting installation structure in the spiral shape microchannel road as described in claim 63, it is characterised in that: repeatedly string
The type of attachment of connection is to be combined with each other multiple tandem spiral shapes microchannel with concatenated type of attachment；Or by tandem spiral
Shape microchannel and microchannel monomer are combined with each other with concatenated type of attachment.
65. the combination connecting installation structure in the spiral shape microchannel road as described in claim 63, it is characterised in that: in parallel
Type of attachment further includes that the n-th cell outlet of (K-1) a microchannel monomer and the n-th of k-th microchannel monomer is thin
The connection convergence of born of the same parents' exit seal is combined into unified cell outlet；By the n-th waste liquid outlet of (K-1) a microchannel monomer with
The n-th waste liquid outlet of k-th microchannel monomer is tightly connected convergence and is combined into unified waste liquid outlet.
66. the combination connecting installation structure in the spiral shape microchannel road as described in claim 63, it is characterised in that: repeatedly simultaneously
The type of attachment of connection is to be combined with each other multiple parallel spiral shape microchannels with type of attachment in parallel；Or by parallel spiral
Shape microchannel is combined with each other with microchannel monomer in form in parallel.
67. the combination connecting installation structure in the spiral shape microchannel road as described in claim 63, it is characterised in that: series-parallel
Combined type of attachment is that one or more parallel spiral shape microchannels and/or one or more tandem spiral shapes are micro- logical
Road and/or one or more microchannel monomers are combined with each other with concatenated type of attachment or the type of attachment of parallel connection, form mixed connection
Formula spiral flow channel.
68. a kind of single-screw shape microchannel, it is characterised in that: including entrance, the microchannel being connect with inlet seal and with it is logical
The outlet that road is tightly connected far from one end of entrance, the entrance is converged by cell entry and sheath fluid entrance, described
It is converged by waste liquid outlet and cell outlet outlet, wherein cell entry is used to enter the sample containing rare cell or particle
Liquid；Cell outlet is used to collect the rare cell or particle after sorting enrichment；Sheath fluid entrance has osmotic pressure for adding, and protects
Demonstrate,prove rare cell or the complete dilution of particle.
69. the single-screw shape microchannel as described in claim 68, it is characterised in that: the cell entry and/or sheath fluid enter
Mouth and/or waste liquid outlet and/or cell outlet are arranged on the outside of the geometric shape of microchannel.
70. a kind of structure that is installed in series of single-screw shape microchannel, which is characterized in that application such as the list in claim 68 or 69
Spiral shape microchannel, in such a way that P single-screw shape microchannel is arranged in series, (Q-1) a single-screw shape microchannel it is thin
Born of the same parents outlet is connected with the cell entry of the Q single-screw shape microchannel, 1 < Q≤P.
71. the structure that is installed in series of the single-screw shape microchannel as described in claim 70, which is characterized in that (Q-1) is a
Pressure balance module is equipped at the waste liquid outlet of single-screw shape microchannel.
72. the structure that is installed in series of the single-screw shape microchannel as described in claim 71, which is characterized in that the pressure is flat
The module that weighs is pressure pump or pressure balance microchannel.
73. the structure that is installed in series of the single-screw shape microchannel as described in claim 72, which is characterized in that the pressure is flat
The module that weighs is the pressure balance microchannel being tightly connected with the waste liquid outlet of the single spiral shape microchannel (Q-1).
74. the structure that is installed in series of the single-screw shape microchannel as described in claim 73, which is characterized in that the pressure
Balance microchannel and matched all single-screw shapes microchannel form non-cross spiral shape microchannel, and one or more
The outlet of a pressure balance microchannel is arranged in outside the geometric figure of spiral shape microchannel, by adjusting pressure balance microchannel
Length and internal diameter adjust the flow velocity in matched cell outlet by content.
75. the structure that is installed in series of the single-screw shape microchannel as described in claim 74, which is characterized in that all pressure are flat
The outlet of weighing apparatus microchannel is arranged at outside the geometric figure of spiral shape microchannel.
76. the structure that is installed in series of the single-screw shape microchannel as described in claim 70, which is characterized in that the cell enters
Mouth and/or sheath fluid entrance and/or waste liquid outlet and/or cell outlet are arranged on the outside of the geometric shape of spiral shape microchannel.
77. a kind of single-screw shape microchannel is installed in parallel structure, which is characterized in that application such as the list in claim 68 or 69
Spiral shape microchannel, in such a way that X single-screw shape microchannel is arranged in parallel, sample solution is from X single-screw shape microchannel
Cell entry enter, rare cell or particle are flowed out from the cell outlet of X single-screw shape microchannel；X > 1.
78. single-screw shape microchannel as described in claim 77 is installed in parallel structure, which is characterized in that by X single spiral shell
The cell entry and/or cell outlet for revolving shape microchannel are interconnected.
79. spiral shape microchannel as described in claim 77 is installed in parallel structure, which is characterized in that in the single spiral shell of X
It revolves and is equipped with pressure balance module at the waste liquid outlet of shape microchannel；The pressure balance module is that pressure pump or pressure balance are micro- logical
Road.
80. spiral shape microchannel as described in claim 79 is installed in parallel structure, which is characterized in that the pressure is flat
Weighing apparatus microchannel and matched single-screw shape microchannel form non-cross spiral shape microchannel, one or more pressure
The outlet of balance microchannel is arranged in outside the geometric figure of spiral shape microchannel, by adjusting the length of pressure balance microchannel
The flow velocity in matched cell outlet by content is adjusted with internal diameter.
81. spiral shape microchannel road is installed in parallel structure as described in claim 77, which is characterized in that all cell entries
And/or sheath fluid entrance and/or waste liquid outlet and/or cell outlet are arranged on the outside of the geometric shape of microchannel.
82. spiral shape microchannel road as described in claim 79 is installed in parallel structure, it is characterised in that: all cells enter
Mouth and/or sheath fluid entrance are arranged at outside the geometric figure of spiral shape microchannel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| PCT/CN2018/122352 WO2019128841A1 (en) | 2017-12-25 | 2018-12-20 | Spiral microchannel, use method thereof, and series/parallel-connected installation structure |
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| CN201711438475.9A CN108132208A (en) | 2017-12-25 | 2017-12-25 | A kind of spiral shape microchannel and its application method and series and parallel installation method |
| CN2017114384759 | 2017-12-25 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106069A (en) * | 2019-05-30 | 2019-08-09 | 合肥工业大学 | A kind of biochip based on fluid dynamic label-free separation cancer cell |
| CN110975775A (en) * | 2019-10-29 | 2020-04-10 | 肇庆市华师大光电产业研究院 | High-flux three-dimensional microfluid mixing device based on dean flow effect and mixing method thereof |
| CN114207407A (en) * | 2019-05-31 | 2022-03-18 | 安伯斯顿生物科学股份有限公司 | Microfluidic determination of low abundance events |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108132208A (en) * | 2017-12-25 | 2018-06-08 | 黄庆 | A kind of spiral shape microchannel and its application method and series and parallel installation method |
| CN108485910A (en) * | 2018-03-21 | 2018-09-04 | 中国人民解放军陆军军医大学第附属医院 | A kind of Double-spiral micro-fluidic chip |
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| WO2023041948A1 (en) * | 2021-09-15 | 2023-03-23 | Fundación Para La Investigación Médica Aplicada | Device suitable for exchanging a liquid medium and selecting particles in a mixture |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103977468A (en) * | 2014-05-26 | 2014-08-13 | 国家纳米科学中心 | System and method for separating and removing circulating tumor cell and blood platelet in blood |
| CN104111190A (en) * | 2014-07-18 | 2014-10-22 | 国家纳米科学中心 | Double-screw micro-fluidic chip |
| US20160241201A1 (en) * | 2013-10-18 | 2016-08-18 | Telefonaktiebolaget Lm Ericsson (Publ) | Power amplifier for amplification of an input signal into an output signal |
| WO2017131580A1 (en) * | 2016-01-28 | 2017-08-03 | Clearbridge Biomedics Pte Ltd | Multi-stage target cell enrichment using a microfluidic device |
| US20170225166A1 (en) * | 2014-08-07 | 2017-08-10 | The General Hospital Corporation | Platelet-Targeted Microfluidic Isolation of Cells |
| CN107058060A (en) * | 2017-04-10 | 2017-08-18 | 东南大学 | A kind of tumour cell high flux sorting enrichment micro-fluidic chip |
| CN107084916A (en) * | 2017-03-29 | 2017-08-22 | 无锡纳奥生物医药有限公司 | A kind of circulating tumor cell separating micro-fluidic chip device and its application method |
| CN209432573U (en) * | 2017-12-25 | 2019-09-24 | 黄庆 | A kind of spiral shape microchannel and its series and parallel mounting structure |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102884170A (en) * | 2010-03-04 | 2013-01-16 | 新加坡国立大学 | Microfluidics sorter for cell detection and isolation |
| JP6314220B2 (en) * | 2013-10-16 | 2018-04-18 | クリアブリッジ バイオメディックス ピーティーイー エルティーディーＣｌｅａｒｂｒｉｄｇｅ Ｂｉｏｍｅｄｉｃｓ Ｐｔｅ Ｌｔｄ | Microfluidic sorter for cell detection and isolation |
| WO2016064896A1 (en) * | 2014-10-20 | 2016-04-28 | University Of Utah Research Foundation | Tissue sample processing system and associated methods |
| CN108132208A (en) * | 2017-12-25 | 2018-06-08 | 黄庆 | A kind of spiral shape microchannel and its application method and series and parallel installation method |
| CN108485910A (en) * | 2018-03-21 | 2018-09-04 | 中国人民解放军陆军军医大学第附属医院 | A kind of Double-spiral micro-fluidic chip |
-
2017
- 2017-12-25 CN CN201711438475.9A patent/CN108132208A/en active Pending
-
2018
- 2018-11-09 CN CN201811331974.2A patent/CN109580323B/en active Active
- 2018-12-20 WO PCT/CN2018/122352 patent/WO2019128841A1/en active Application Filing
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160241201A1 (en) * | 2013-10-18 | 2016-08-18 | Telefonaktiebolaget Lm Ericsson (Publ) | Power amplifier for amplification of an input signal into an output signal |
| CN103977468A (en) * | 2014-05-26 | 2014-08-13 | 国家纳米科学中心 | System and method for separating and removing circulating tumor cell and blood platelet in blood |
| CN104111190A (en) * | 2014-07-18 | 2014-10-22 | 国家纳米科学中心 | Double-screw micro-fluidic chip |
| US20170225166A1 (en) * | 2014-08-07 | 2017-08-10 | The General Hospital Corporation | Platelet-Targeted Microfluidic Isolation of Cells |
| WO2017131580A1 (en) * | 2016-01-28 | 2017-08-03 | Clearbridge Biomedics Pte Ltd | Multi-stage target cell enrichment using a microfluidic device |
| CN107084916A (en) * | 2017-03-29 | 2017-08-22 | 无锡纳奥生物医药有限公司 | A kind of circulating tumor cell separating micro-fluidic chip device and its application method |
| CN107058060A (en) * | 2017-04-10 | 2017-08-18 | 东南大学 | A kind of tumour cell high flux sorting enrichment micro-fluidic chip |
| CN209432573U (en) * | 2017-12-25 | 2019-09-24 | 黄庆 | A kind of spiral shape microchannel and its series and parallel mounting structure |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106069A (en) * | 2019-05-30 | 2019-08-09 | 合肥工业大学 | A kind of biochip based on fluid dynamic label-free separation cancer cell |
| CN114207407A (en) * | 2019-05-31 | 2022-03-18 | 安伯斯顿生物科学股份有限公司 | Microfluidic determination of low abundance events |
| CN110975775A (en) * | 2019-10-29 | 2020-04-10 | 肇庆市华师大光电产业研究院 | High-flux three-dimensional microfluid mixing device based on dean flow effect and mixing method thereof |
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|---|---|
| CN109580323B (en) | 2024-07-19 |
| CN108132208A (en) | 2018-06-08 |
| WO2019128841A1 (en) | 2019-07-04 |
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