CN108132208A - A kind of spiral shape microchannel and its application method and series and parallel installation method - Google Patents
A kind of spiral shape microchannel and its application method and series and parallel installation method Download PDFInfo
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Abstract
The invention belongs to rare cell or particle rich screening fields,It is related to a kind of spiral shape microchannel and its application method and string,It is installed in parallel method,For technical barrier of the spiral shape microchannel in the sorting enrichment field of Peripheral Blood from Patients with Malignant circulating tumor cell,Dexterously have devised the series connection of the multiple spiral shape microchannels of reasonable application of the manystage cascade connection and parallel spiral shape microchannel inertial separation structure,Relative to existing similar technique,The present invention is enriched with Peripheral Circulation tumour cell to sorting,Not only there is 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,The cell suspension containing high-purity circulating tumor cell obtained is particularly suitable for subsequent biomedical detection,Genetic analysis,Cell culture,Xenograft tumor preparation etc.,With extensive and far-reaching clinical practice application value.
Description
Technical field
The invention belongs to rare cell or particle rich screening fields, are related to a kind of spiral shape microchannel and its application method
With series and parallel installation method.
Background technology
Invasion and transfer are the Major Clinical causes of death of tumor patient, and the death caused by metastases disease accounts for entity
Dead 90% or so caused by knurl, therefore, it is important that early stages of metastases, precisely detection, which seemed excellent,.Circulating tumor cell refers to
Primary and metastatic tumour lesion is detached from, invades and enter the tumour cell of the circulatory systems such as lymphatic vessel, blood.It is examined in the circulatory system
The presence of tumor micrometastasis may be had, and make further progress as the possibility of DISTANT METASTASES IN by measuring circulating tumor cell prompting
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, is referred to as liquid biopsy, in recent years into
The hot spot precisely diagnosed for tumour.
Circulating tumor cell in the circulatory system not only has morphology and science of heredity heterogeneity, 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, realize circulating tumor cell
Detection, it is an essential step that sorting enrichment is carried out to it, also, the good and bad of circulating tumor cell sorting enrichment will
Directly affect its subsequent detection result (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
With one of method, it is mainly characterized by that any external force need not be applied, such as electric power, magnetic force, it is possible to realize and include in microchannel
The focusing flowing of certain size particle including cell.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 of curved arc channel.When fluid is laminar state in linear path, one kind is presented in the flow velocity of fluid
Parabolic shape is distributed：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 near enough, channel
Particle can be pushed away conduit wall by the lift that wall induction generates again.The resultant force of the opposite lift in above two direction is referred to as inertia liter
Power FL.Inertia lift is acted 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 so as to which particle be made to focus on settling position in cross-section, forms and focuses on flowing, flow to downstream.Particle is in linear
The position of stress balance is related with the shape of microchannel cross-section in microchannel cross-section：In the microchannel of circular cross section,
Particle can focus on to form an annular；In the microchannel of square cross section, the position that particle focuses on 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 on can be reduced to two, respectively positioned 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, so as 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, so as to be squeezed 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, respectively positioned at the top of microchannel cross-section and
Thus lower part generates a kind of Secondary Flow referred to as Dien vortex.Dien vortex can generate the particle in fluid drag force effect,
It is 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 was both above-mentioned with curved arc shape microchannel
Focus on 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 in the particle of outer side wall by the microchannel transverse direction of certain length
When being moved to madial wall, 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, it is referred to as completing a complete Dien period, in other words, completes 1 enlightening
The grace period.As described above, other than Dien drag force, larger particle (particularly its diameter reaches a/h >=0.07) can also be simultaneously
It is acted on by inertia lift, so as to which the focal position for making particle is very dependent on the ratio (F of inertia lift and Dien drag forceL/
FD), the size of above-mentioned ratio and particle has exponent relation, and the focusing of particle depends on minimum microchannel scale parameter
(the height h) of such as spiral shape microchannel.Larger particle is (i.e.：A/h >=0.07), Dien drag force and inertia lift are in one
Equilibrium state can focus on the madial wall of spirality channel, and in the microchannel in downstream behind the microchannel of certain length
Remain at the madial wall of microchannel；And smaller particless are (i.e.：a/h<0.07), Dien drag force is dominant, and particle is with Dien
Eddy currents are constantly traversing back and forth between the lateral wall and madial wall 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.On this condition, the sheath fluid fluid with fair speed can squeeze particle flux to microchannel
Lateral wall；When particle flows in microchannel, the particles of all original states (i.e. inlet) is in the effect of Dien drag force
Under, madial wall is moved to by micro-tunnel length of the lateral wall through 0.5 Dien period；After nearly madial wall is reached, larger grain
Son is (i.e.：A/h >=0.07) brute force focussing is in madial wall when by strong inertia lift, and further in the work of Dien drag force
It is prevented further to be moved 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, larger particle focuses on the madial wall of microchannel always, and is remained in subsequent downstream microchannel
The focused flow of madial wall in microchannel；But it is moved to the smaller particless of madial wall (i.e. through 0.5 Dien period： 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 madial wall.As it can be seen that larger particle is always
Madial wall in spiral shape microchannel focuses on a fluid stream in inertia, and smaller particless are then with Dien eddy currents in the outside of microchannel
Wall and madial wall lateral transfer back and forth.When the spiral shape microchannel of design specific length, exit just makes smaller particle
In or near lateral wall, so that it may larger particle is detached with smaller particless by appropriate outlet, realizes 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 to the red blood cell (diameter is about 6-8 μm) in peripheral blood and leucocyte, (diameter is about
8-12 μm), the size bigger of circulating tumor cell (diameter is about 20-30 μm), by limiting the rectangular cross section of aspect ratio,
Under particular fluid flow conditions, Peripheral Circulation tumour cell will flow to cell along the madial wall focus layer of spiral shape microchannel
It exports, remaining cell in sample is thin so as to fulfill circulating tumor then along the lateral wall laminar flow of the channel to waste liquid outlet
The separation of born of the same parents.Meanwhile it can also make there is 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
It is that obtained circulating tumor cell purity is low to want drawback, should with clinical practice so as to cause follow-up differentiate of circulating tumor cell
With there are bigger difficulty, practical clinical value is relatively low.Although existing spiral shape microchannel is in sorting enrichment cycles
Larger deficiency and relevant technical bottleneck are still had in terms of tumour cell, still, existing result of study prompting, if energy
Enough dexterously " cascade " applies multiple single spiral shape microchannels, just 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 that greatlys improve is enriched to makes it meet clinical reality
Border demand.
Invention content
In view of this, the purpose of the present invention is to provide a kind of spiral shape microchannels and its application method to pacify with series and parallel
Dress method.
In order to achieve the above objectives, the present invention provides following technical solution：
A kind of spiral shape microchannel, it is single including N number of single-screw shape microchannel for sorting enrichment rare cell or particle
A single-screw shape microchannel includes entrance, the microchannel being connect with inlet seal and is sealed with the one end of channel far from 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 is sealed with the cell entry of m-th single-screw shape microchannel
(M-1) a classification cycle point is connected into, it is micro- by the waste liquid outlet of (M-1) a single-screw shape microchannel, m-th single-screw shape
The sheath fluid entrance of channel and (M-1) a classification cycle point are formed 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 put down 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 for the liquid flow direction and for making 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 liquid flow direction of the cell outlet of a single-screw shape microchannel and the cell outlet of (M-1) 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 liquid flow direction of mouth and the waste liquid outlet of (M-1) a single-screw shape microchannel 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 for the rare cell or particle after collection sorting enrichment；The sheath fluid enters
Mouth ensures rare cell or the complete dilution of particle for adding with osmotic pressure；1 ＜ M≤N, M, N is positive integer.
Optionally, the pressure balance module is a single spiral shells of (M-1) being arranged at (M-1) a classification cycle
The pressure balance microchannel revolved at the waste liquid outlet of shape microchannel and be tightly connected with the waste liquid outlet；The pressure balance is micro- logical
It is a that road is equipped with (N-1).
Optionally, the pressure balance microchannel is set in the same plane with N number of single-screw shape microchannel.
Optionally, the pressure balance module is a single spiral shells of (M-1) being arranged at (M-1) a classification cycle
Revolve the compression 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 mutually parallel 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 rectangular cross-section or trapezoidal of single single-screw shape microchannel.
Optionally, the rectangular cross-section of the single-screw shape microchannel, the ratio w/ of the width w and height h of the rectangle
H=1.2~10.
Optionally, the section of the single-screw shape microchannel is trapezoidal, the trapezoidal close spiral shape microchannel
The side height of geometric center is less than the side height of 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 forms.
Optionally, the distance between outlet of the entrance of single single-screw shape microchannel and 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
The Dien period.
Optionally, the channel cross-section width maximum width of the single-screw shape microchannel is l1, and the single-screw shape is micro- logical
The width of the cell entry in road is l2,0.1*l1≤l2≤0.4*l1；The width of the waste liquid outlet of the single-screw shape microchannel is
L3,0.4*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, a diameter of a of the rare cell or particle, the height of the microchannel are h, 0.07≤a/h≤
0.5。
Optionally, the rare cell or particle are circulating tumor cell, cycle progenitor cells, recycle fetal cell, cell
Vesica, excretion body or pathogen.
Optionally, can the cell outlet of n-th single-screw shape microchannel set detection unit, realize cell outlet at pair
It sorts the cell being enriched to or particle is identified and/or counted；Bio-light well known to the technical field of the invention can be used
, electrobiology, bioacoustics and/or biomagnetism technology realize the property of cell or particle in detection unit of the present invention
Shape identify, including but be not intended to limit THz wave spectral technology, Raman spectroscopy, electrical impedance technology, differential impedance or difference resistance
Anti- spectral technology.It, can be by cell or particle too it is characterized in that when the cell that is enriched to of sorting or particle pass through detection unit
The features such as hertz wave spectrum, Raman spectrum, impedance, differential impedance identify the character of cell or particle.
A kind of application method of spiral shape microchannel, includes 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：Cell outlet in n-th single-screw shape microchannel collects the fluid rich in rare cell or particle；First
The waste liquid outlet of the waste liquid outlet of a single-screw shape microchannel to n-th single-screw shape microchannel discharges waste liquid.
Optionally, the sample liquid used in step S1 is for the blood sample with certain hematid specific volume or by molten
Blood 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.
Optionally, step S1.1 is further included,
S1.1：Adjust pressure balance module, make m-th single-screw shape microchannel cell entry 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 cell outlet and the cell of (M-1) a single-screw shape microchannel of m-th single-screw shape microchannel go out
The liquid flow direction of mouth is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Make the useless of m-th single-screw shape microchannel
The liquid flow direction of the liquid flow direction of liquid outlet and the waste liquid outlet of (M-1) a single-screw shape microchannel is consistent, and
0.7*v6≤v5≤1.3*v6。
A kind of method that is installed in series of spiral shape microchannel, by the way of being arranged in series using P spiral shape microchannel, the
(Q-1) first of the cell outlet of the n-th single-screw shape microchannel of a spiral shape microchannel and the Q spiral shape microchannel
Single-screw shape microchannel entrance is connected, and forms spiral shape microchannel and is installed in series place, 1 ＜ Q≤P.
Optionally, be installed in series in spiral shape microchannel place (Q-1) a spiral shape microchannel 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 of channel is tightly connected.
Optionally, the pressure balance module is to be arranged on spiral shape microchannel to be installed in series (Q-1) a spiral at place
Compression 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, by the way of being arranged in series using 2 spiral shape microchannels, 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 method, by the way of being arranged in parallel using X spiral shape microchannel, 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 outflow of the n-th single-screw shape microchannel of spiral shape microchannel；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 set 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 plane where single spiral shape microchannel
Direction is set.
Optionally, X=2, by the way of being arranged in parallel using two spiral shape microchannels, sample solution is micro- from two spiral shapes
The cell entry of first single-screw shape microchannel of channel enters, and rare cell or particle are from the of two spiral shape microchannels
The cell outlet outflow of N number of single-screw shape microchannel.
Optionally, N=2, by the way of being arranged in parallel using X Double-spiral microchannel, sample solution is from X double helix
The cell entry of first single-screw shape microchannel of shape microchannel enters, and rare cell or particle are micro- logical from X Double-spiral
The cell outlet outflow of second single-screw shape microchannel in road.
The beneficial effects of the present invention are：
The present invention is enriched with field for sorting of the spiral shape microchannel in Peripheral Blood from Patients with Malignant circulating tumor cell
Technical barrier, 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, capture rate and processing flux not only with good circulating tumor cell, it is also possible to
The purity of circulating tumor cell is maximized, 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 flow, obtained containing 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 the feature of the present 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.The target and other advantages of the present invention can by following specification realizing and
It obtains.
Description of the drawings
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, wherein：
Fig. 1 is the structure diagram of embodiment one in the present invention；
Fig. 2 is the structure diagram of embodiment two in the present invention；
Fig. 3 is the structure diagram of embodiment three in the present invention；
Fig. 4 is the structure diagram of example IV in the present invention；
Fig. 5 is the structure diagram of embodiment five in the present invention；
Fig. 6 is the structure diagram of embodiment six in the present invention；
Fig. 7 is the structure diagram of embodiment seven in the present invention；
Fig. 8 is movement locus figure of the various sizes of particle under different in flow rate in the embodiment of the present invention one；
Fig. 9 is the microscope photograph in different spiral shape microchannels exit in the embodiment of the present invention one；
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.
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
Disclosed content understands other advantages and effect of the present invention easily.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 be based on different viewpoints with application, without departing from
Various modifications or alterations are carried out under the spirit of the present invention.It should be noted that diagram provided in following embodiment is only to show
Meaning mode illustrates the basic conception of the present invention, and in the absence of conflict, the feature in following embodiment and embodiment can phase
Mutually combination.
Please refer to Fig.1-Figure 11, the element numbers in attached drawing represent respectively：The cell of first single-screw shape microchannel enters
The entrance A of 2, first single-screw shape microchannels of sheath fluid entrance of 1, first single-screw shape microchannel of mouth, pressure balance are micro- logical
5, first lists of sheath fluid entrance of 4, second single-screw shape microchannels of cell outlet of 3, the first single-screw shape microchannels in road
The outlet B of 6, first single-screw shape microchannels of waste liquid outlet of spiral shape microchannel, second single-screw shape microchannel it is useless
Liquid exports outlet C, second spiral shell of 8, second single-screw shape microchannels of cell outlet of 7, second single-screw shape microchannels
Revolve first single spiral shell 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 rotation shape microchannel,
The second of 12, second spiral shape microchannels of cell outlet of second single-screw shape microchannel of second spiral shape microchannel
The cell of first single-screw shape microchannel of 13, second spiral shape microchannels of waste liquid outlet of a single-screw shape microchannel enters
Mouthfuls 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
The sheath fluid entrance 17 of single-screw shape microchannel, the cell outlet 18 of the 4th single-screw shape microchannel, the 4th single-screw shape are micro-
The waste liquid outlet 19 of channel.
Embodiment one
The present invention relates to a kind of spiral shape microchannels, for sorting enrichment rare cell or particle, wrap in the present embodiment
Two single-screw shape microchannels are included, which sets in the same plane and non-crossly wind cyclization
Shape, as shown in Figure 1.
Single single-screw shape microchannel include entrance A, the microchannel that is connect with inlet seal and with channel it is separate 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 entry sealing of the cell outlet 4 of first single-screw shape microchannel and second single-screw shape microchannel connects
First classification cycle point is connected into, 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 formed 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
Road 3, the flow rate of liquid of 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 be 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 is 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
The liquid flow direction of the cell outlet 8 of two single-screw shape microchannels and the cell outlet 4 of first single-screw shape microchannel
Liquid flow direction it is consistent, and 0.7*v4≤v3≤1.3*v4, and v1=v4；Make the useless of second single-screw shape microchannel
The liquid flow direction of the liquid flow direction of liquid outlet 7 and the waste liquid outlet 6 of first single-screw shape microchannel 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 for the rare cell or particle after collection 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 ooze for adding to carry
Pressure thoroughly ensures rare cell or granule integrity and the dilution of biological activity.
Using the form of two single-screw shape microchannels, second single-screw shape microchannel can be directly to first list spiral shell
The rare cell or particle that the sorting of rotation shape microchannel is enriched to carry out further sorting enrichment, so as to than single single-screw shape
Microchannel captures the more efficient of rare cell or particle, be equivalent to rare cell or particle have been 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
Through be directly connected to from beginning to end mode form an appearance for it is parallel and with identical centre point Double-spiral microchannel entirety it is 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 of 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 madial wall 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 or madial wall height of ratio be less than lateral wall height trapezoid cross section, for because target cell it is thin with non-target
Size difference existing for born of the same parents and only make target cell microchannel madial wall formed focused flow, non-target cell is then with Dien vortex-like fashion
The lateral transfer back and forth between microchannel madial wall and lateral wall.Further, the width of microchannel is 0.1-1000 μm, depth
It it is 0.1-800 μm or further, the width of microchannel is 100-800 μm, and depth is 50-500 μm or further, 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.On 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 the 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 with direction holding or extremely close, meanwhile, make flow velocity and the direction of the waste liquid outlet 6 of first single-screw shape microchannel
It is identical with direction holding or extremely close with the flow velocity of the waste liquid outlet 7 of second single-screw shape microchannel.
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 of 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 there is 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 inertia in curved arc shape microchannel and focus on flowing and the size of microchannel and radius of curvature, particle
Size and the factors such as flow velocity have relationship.Existing research shows that channel width be more than height rectangular section channel
In, the particle of certain size forms inertia and focuses on flowing it is generally necessary to meet the diameter that condition a/h >=0.07, a are particles, and h is
The minimum dimension of microchannel, the height of microchannel is typically minimum dimension in inertia micro flow chip.It, can if particle size is too small
Particle can be caused inertia can not to be focused 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 easily blocks.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 enrichment Peripheral Circulation tumour cell is sorted from peripheral blood cascade or tandem superposition
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
The sheath fluid entrance 2 of a single-screw shape microchannel and 5 sample introduction of sheath fluid entrance of second single-screw shape microchannel, in certain fluid
Under flow conditions, the cell outlet 8 second single-screw shape microchannel collects the fluid 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 operation principle of Double-spiral microchannel of the present invention is as follows：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
Put 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 madial wall first single-screw shape microchannel, which is formed, focuses on 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 madial wall；Non-target cell is mainly acted on because its size is smaller by Dien drag force, with Dien vortex
Form is in the lateral wall of first single-screw shape microchannel and madial wall lateral transfer back and forth, also, typically 0.5 Dien week
Positioned at the madial wall of first single-screw shape microchannel during 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 positioned at first single-screw shape microchannel madial wall
The cell entry of second single-screw shape microchannel is directly entered, and non-target cell fluid is then from micro- positioned at first single-screw shape
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 operation 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 by the 2nd sorting enrichment or tandem type sorting enrichment or tandem sorting enrichment or superposing type
Sorting enrichment.It can thus be stated that by Double-spiral microchannel of the present 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 removes, 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, so as to, because the interference of its density is lacked, sort concentration effect meeting due to cell
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 exports, wherein, the width of cell entry and cell outlet is 0.15mm, respectively positioned at the outer of spiral shape microchannel
The width of side wall and madial wall, sheath fluid entrance and waste liquid outlet is 0.45mm, respectively positioned at the madial wall of spiral shape microchannel 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
Outlet 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 both order spiral shells
Revolve 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, gives up
Liquid outlet 6 is located at the exterior sidewall portions of set single-screw shape microchannel outlet；The cell outlet of first single-screw shape microchannel
4 and the cell entry of second single-screw shape microchannel be tightly connected into first classification cycle point；By first single-screw shape
The sheath fluid entrance 5 of 6, second single-screw shape microchannels of waste liquid outlet of microchannel and first classification cycle point form first
At a classification cycle；The waste liquid outlet 6 of first single-screw shape microchannel at first classification cycle is put down equipped with pressure
Weighing apparatus microchannel 3.It is finally generated to different size cells in view of two factors of variation different in flow rate and passage length
The difference of lateral displacement focal position, final optimization pass determine optimal parameter.
The making 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 chip, to being carried out at punching at its corresponding entrance after PDMS demouldings
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 mode of magnetic fixture device and accurate guide pin
It is closely connect with disengaging micro-pipe, and passes through syringe pump and flow velocity control system so as to reach sample introduction stabilization and can supervise in real time
The purpose of velocity measurement variation.Before its separating effect test 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 the connection of chip entrance compared with
Securely, by micro- Microscopic 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 simulates red blood cell, leucocyte, circulating tumor cell respectively, using phosphorus
Phthalate buffer is diluted to 1.0*10 respectively to 3~6 μm, 7~10 μm, 15~18 μm of microballoon7Particle/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 to by accurate guide pin on the outside of the spiral shape chip after excluding bubble
With madial wall entrance, by flow rate monitoring system and compression pump control adjustment flow velocity, record and collect of the present invention respectively
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 detaches at cell outlet is shown in Fig. 8, and list is captured 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 movement locus, with compared with
The dot of minor diameter represents 3~6 μm and 7~10 μm of rare cell or particle, and 15~18 μm are represented with larger-diameter dot
Rare cell or particle movement locus.The microscopic counting observation knot that microballoon detaches at cell outlet 8 and waste liquid outlet 7
Fruit sample situation is shown in Fig. 9.Fig. 9-a be traditional single-screw shape microchannel cell outlet at displaing micro picture, three kinds of microballoons compared with
More, the sorting concentration effect for prompting larger particle is poor, and purity is low；Fig. 9-b are Double-spiral microchannel of the present invention cell
The displaing micro picture in exit is almost larger-diameter microballoon, and the sorting concentration effect for prompting larger particle is very good, pure
It spends high；Fig. 9-c are the displaing micro picture at Double-spiral microchannel of the present invention waste liquid outlet 7, it is seen that are almost smaller
The microballoon of diameter almost loses larger-diameter microballoon, and larger particle is prompted to be enriched to cell outlet, has good time
Yield.Further analysis result shows that, when flow velocity is in 600~750 μ l/min, two kinds of spiral shape microchannels are respectively provided with most preferably
Sorting 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 inside
The cell outlet of wall；When the flow velocity of fluid is more than 750 μ l/min, the cell purity of larger particle declines；Associated exemplary result
As shown in Figure 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
The large-size particle of road sorting enrichment is (i.e.：15~18 μm) purity and repeatability to be far superior to traditional single-screw shape micro- logical
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 so that lateral displacement occurs for the smaller particle of size, so as to the big ruler for being enriched to sorting
Very little particle is (i.e.：15~18 μm) repeatability of purity and experimental result is affected.But Double-spiral of the present invention is micro-
Channel because can two level sorting enrichment, so as to 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
During liquid sample, traditional single-screw shape microchannel takes 40 minutes, and Double-spiral microchannel of the present invention only takes 10 points
Clock, and the purity of circulating tumor cell that is obtained of Double-spiral microchannel of the present invention and the repeated higher of experimental result
(close with result described in table 1), the two have statistically-significant difference.The studies above result is prompted, relative to traditional single spiral shell
Shape microchannel is revolved, Double-spiral microchannel of the present invention more can meet and meet the clinical practice detection need of circulating tumor cell
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 1 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,
According to 1:5 volume ratio is mixed into 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 2 using the count results of immunofluorescence technique stain smear.As a result this is shown
It invents the Double-spiral microchannel and more than 98%, and purity and above-mentioned double spiral shells is at least up to the rate of recovery of kinds of tumor cells
It is similar to revolve shape microchannel, is each about 99%.
2 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 the rare cell that is enriched to of sorting or the purity of 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 practice.
Embodiment two
The present embodiment in the present embodiment using impressed pressure pump what is different from the first embodiment is that realized with 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
The liquid of the liquid flow direction of the cell outlet 8 of single-screw shape microchannel and the cell outlet 4 of first single-screw shape microchannel
Body 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 liquid flow direction of mouth 7 and the waste liquid outlet 8 of first single-screw shape microchannel is consistent, and 0.7*v6
≤v5≤1.3*v6.The compression 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 spiral shells
Revolve 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 microchannels, or more grade spiral shape microchannel.Included single-screw shape is micro- in the multiple coil shape microchannel
The quantity of channel is at least 2 or more, also, the center of circle of the multiple single-screw shape microchannel is located at same point, and (M-1) is a
The cell outlet of single-screw shape microchannel is the cell entry of m-th single-screw shape microchannel, and (M-1) a single-screw shape is micro-
The waste liquid outlet of channel has been directly connected to pressure equalisation passage.The effect of the pressure equalisation passage is to make (M-1) a single spiral shell
The flow velocity and direction of the cell outlet of rotation shape microchannel or the cell entry of m-th single-screw shape microchannel and (M-1) a spiral shell
The flow velocity and direction for revolving the cell entry of shape microchannel and the cell outlet of m-th single-screw shape microchannel keep identical or pole phase
Closely, meanwhile, make m-th single-screw shape microchannel waste liquid outlet flow velocity and direction and (M-1) a spiral shape microchannel it is useless
The flow velocity of liquid outlet and direction keep identical or extremely close, so as to make each spiral shape in the multiple coil shape microchannel micro-
The hydrodynamics characteristic that channel gives target cell 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
Revolve the cell entry of shape microchannel, 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 respectively has corresponding pressure
Balance channel 3.
The sheath fluid is from the sheath fluid of 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
The rare cell or particle after sorting enrichment are collected at the cell outlet 18 of four single-screw shape microchannels.Example IV
The present embodiment is realized and pressure in embodiment unlike embodiment three in the present embodiment using impressed pressure pump
Balance the approximate effect in microchannel.
Embodiment five
In the present embodiment, two spiral shape microchannels are connected by the way of series connection, the of first spiral shape microchannel
The cell outlet 8 of two single-screw shape microchannels and the cell of first single-screw shape microchannel of second spiral shape microchannel
Entrance 14 is connected, and forms the place that is installed in series of two spiral shape microchannels, and the place that is installed in series in each spiral shape microchannel
Position on pressure equalisation passage is set.
The sheath fluid is from the sheath fluid entrance 2, first of first single-screw shape microchannel of first spiral shape microchannel
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 9 of shape microchannel and the sheath fluid entrance of second single-screw shape microchannel of second spiral shape microchannel
11 import；Waste liquid is from 6, first spiral shapes of waste liquid outlet of 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 of channel and second spiral shape microchannel is led
Go 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, with showing
Some cell entries and cell outlet compare (example respectively positioned at the inner side and outer side of single spiral flow channel planar geometry
Such as：The last one attached drawing), the present invention can under conditions of without using any external micro-pipe directly cascade or tandem using multiple
N grades of spiral flow channels, both simplify manufacture craft, also make the complexity of N grades of spiral flow channels reduce, and improve working efficiency and inspection
Survey the accuracy of result.
Embodiment six
The present embodiment is realized using impressed pressure pump with being pressed in embodiment five unlike embodiment five in the present embodiment
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 of channel flows out.Participate in the center of circle projection of Double-spiral microchannel in parallel
It overlaps.2,3 or more samples to be detected can be handled simultaneously on same micro-fluidic chip, so as to improve detection effect
The processing time of testing cost and sample is effectively reduced while rate.
Finally illustrate, the above embodiments are merely illustrative of the technical solutions of the present invention and it is unrestricted, although with reference to compared with
The present invention is described in detail in good embodiment, it will be understood by those of ordinary skill in the art that, it can be to the skill of the present 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
Right in.
Claims (38)
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, single single-screw shape microchannel includes entrance, the microchannel being connect with inlet seal
And the outlet being tightly connected with one end far from entrance of channel, the entrance by cell entry and the convergence of sheath fluid entrance and
Into the outlet is converged by waste liquid outlet and cell outlet；
The cell outlet of (M-1) a single-screw shape microchannel is tightly connected with the cell entry of m-th single-screw shape microchannel
Into (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 form 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 is v6；
The pressure balance module is used for the liquid flow direction and (M-1) that make 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 the 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 for the rare cell or particle after collection sorting enrichment；The sheath fluid entrance is used for
Addition ensures rare cell or the complete dilution of particle with osmotic pressure；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 is arranged on 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 is arranged on the
(M-1) at a classification cycle in (M-1) a single-screw shape microchannel waste liquid outlet at compression 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 positioned 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 mutually parallel.
7. spiral shape microchannel as described in claim 6, it is characterised in that：Single spiral shell between the spiral shape microchannel
It is 0.1~1000 μm to revolve the horizontal distance between shape microchannel and single-screw shape microchannel adjacent thereto and parallel.
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- less 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,
Highly it 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 the 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 the single-screw shape microchannel is cut
Face width maximum width is 11, 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 sheath fluid entrance of the single-screw shape microchannel and/or the width of waste liquid outlet are l3,0.4*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 forms.
16. spiral shape microchannel as described in claim 1, it is characterised in that：The rare cell or particle it is a diameter of
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 cycle
Oncocyte, cycle progenitor cells, cycle fetal cell, cell vesicle, excretion body or pathogen.
18. spiral shape microchannel as described in claim 1, it is characterised in that：It further includes 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 on 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 cells or particle；The particular technology means are selected from by Photobiology, electrobiology, biology
The group of acoustics, biomagnetism and its arbitrary combination 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 its arbitrary combination form.
21. the application method of the spiral shape microchannel as described in any one of claim 1~20, which is characterized in that
Include the following steps：
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：Cell outlet in n-th single-screw shape microchannel collects the fluid rich in rare cell or particle；
Waste liquid outlet in waste liquid outlet to the n-th single-screw shape microchannel of first single-screw shape microchannel discharges waste liquid.
22. the application method of the spiral shape microchannel as described in claim 21, 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 samples of a large amount of red blood cells is removed by haemolysis mode
This；Hematid specific volume in the blood sample is 0%~70%.
23. the application method of the spiral shape microchannel as described in claim 21, which is characterized in that is used in step S1 is dilute
Liquid is released with certain osmotic pressure, is phosphate buffer, culture serum or physiological saline for protecting haemocyte integrality.
24. the application method of the spiral shape microchannel as described in claim 21, which is characterized in that step S1.1 is further included,
S1.1：Adjust pressure balance module, make the m-th single-screw shape microchannel cell entry 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 liquid flow direction of outlet and the waste liquid outlet of (M-1) a single-screw shape microchannel is consistent, and
0.7*v6≤v5≤1.3*v6。
25. the method that is installed in series of the spiral shape microchannel as described in claim 1~20, which is characterized in that using 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 (Q-1) a spiral shape microchannel
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.
26. the method that is installed in series of spiral shape microchannel as recited in claim 25, 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 (Q-1) a spiral shape microchannel at the road place of being installed in series
Dynamic balance module.
27. the method that is installed in series of the spiral shape microchannel as described in claim 26, which is characterized in that the pressure balance
Module be with spiral shape microchannel be installed in series place (Q-1) a spiral shape microchannel n-th single-screw shape microchannel
The pressure balance microchannel that waste liquid outlet is tightly connected.
28. the method that is installed in series of the spiral shape microchannel as described in claim 26, which is characterized in that the pressure balance
Module be arranged on spiral shape microchannel be installed in series place (Q-1) a spiral shape microchannel n-th single-screw shape it is micro- logical
Compression pump at the waste liquid outlet in road.
29. the method that is installed in series of spiral shape microchannel as recited in claim 25, which is characterized in that N=2, the spiral shell
It is Double-spiral microchannel to revolve shape microchannel.
30. the method that is installed in series of spiral shape microchannel as recited in claim 25, 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.
31. spiral shape microchannel as described in claim 1~20 is installed in parallel method, which is characterized in that using X spiral shell
The mode that rotation shape microchannel is arranged in parallel, sample solution are thin from first single-screw shape microchannel of X spiral shape microchannel
Born of the same parents' entrance enters, and rare cell or particle are from the cell outlet stream of the n-th single-screw shape microchannel of X spiral shape microchannel
Go out；X ＞ 1.
32. spiral shape microchannel as described in claim 31 is installed in parallel method, which is characterized in that by X spiral shape
The cell entry of first single-screw shape microchannel of microchannel is interconnected.
33. spiral shape microchannel as described in claim 31 is installed in parallel method, which is characterized in that by X spiral shape
The cell outlet of the n-th single-screw shape microchannel of microchannel is interconnected.
34. spiral shape microchannel as described in claim 31 is installed in parallel method, which is characterized in that the X spiral shell
Revolve the setting of shape microchannel in the same plane.
35. spiral shape microchannel as described in claim 33 is installed in parallel method, which is characterized in that the X spiral shell
Rotation shape microchannel is arranged concentrically in the same plane.
36. spiral shape microchannel as described in claim 31 is installed in parallel method, which is characterized in that the X spiral shell
Shape microchannel is revolved with one heart and along perpendicular to the setting of the direction of plane where single spiral shape microchannel.
37. spiral shape microchannel as described in claim 32 is installed in parallel method, which is characterized in that X=2, using two
The mode that a spiral shape microchannel is arranged in parallel, sample solution is from first single-screw shape microchannel of two spiral shape microchannels
Cell entry enter, the cell of rare cell or particle from the n-th single-screw shape microchannel of two spiral shape microchannels goes out
Mouth outflow.
38. spiral shape microchannel as described in claim 32 is installed in parallel method, 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 from the thin of second single-screw shape microchannel of X Double-spiral microchannel
Born of the same parents export outflow.
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Cited By (7)
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| 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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Family Cites Families (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SG183468A1 (en) * | 2010-03-04 | 2012-09-27 | Univ Singapore | Microfluidics sorter for cell detection and isolation |
| WO2015057159A1 (en) * | 2013-10-16 | 2015-04-23 | Clearbridge Biomedics Pte Ltd | Microfluidics sorter for cell detection and isolation |
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- 2018-12-20 WO PCT/CN2018/122352 patent/WO2019128841A1/en active Application Filing
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| WO2019128841A1 (en) | 2019-07-04 |
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