CN109622076A - A kind of 3D printing offers the micro-fluidic chip and the wherein design method of microchannel of sinuous microchannel - Google Patents
A kind of 3D printing offers the micro-fluidic chip and the wherein design method of microchannel of sinuous microchannel Download PDFInfo
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- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
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Abstract
The micro-fluidic chip and the wherein design method of microchannel of sinuous microchannel are offered the invention discloses a kind of 3D printing；The chip is provided with microchannel on substrate, and microchannel is connected in series by multiple circular arcs, so that the channel can realize focusing in the flow rates of 480-720 μ L/min, flow velocity is larger, and flux is high, and then realizes the focusing of particle in large scale channel；The channel design is simple, and 3D printing is easily achieved, and only needs a step that the printing of micro-fluidic chip and 3 D stereo microchannel can be completed, simplifies the production process of micro-fluidic chip；Cost is lower compared with existing micro-fluidic chip manufacturing technology；The design method of the microchannel passes through three step of theoretical calculation, analogue simulation and experimental verification, and the microchannel radius of curvature of optimization is 5.9mm, and when flow velocity is 480 μ L/min, particle or tumour cell focus most close particle in channel.
Description
[technical field]
The invention belongs to micro fluidic chip technical fields, and in particular to a kind of 3D printing offers sinuous microchannel
The design method of micro-fluidic chip and wherein microchannel.
[background technique]
Microflow control technique can carry out biological sample processing on micro-meter scale even individual cell level, be to traditional biological
Major reform of experiment.The microchannel inertia particle focusing of micro-fluidic chip is that one of biomedical applications field is important
Technology has great potential in the fields such as particle or cell focusing, circulating tumor cell separation and flow cytometry.Traditional
Microfluidic channel is manufactured by the soft lithography based on PDMS micro shaping.PDMS is cheap, bio-compatible and transparent,
Using silicon wafer template construct microchannel, precision is high.But this technology production micro-fluidic chip needs a large amount of manual operation,
And the channel of plane can only be made.These hinder soft lithography answering in production complicated three-dimensional channel and business
With.
In past ten years, with the development of 3D printing technique, it is wide that the micro-fluidic core technology of 3D printing manufacture causes people
General concern.The process that micro-fluidic chip can significantly be simplified using 3D printing technique manufacture micro-fluidic chip, is reduced micro-
The cost of manufacture of fluidic chip, while 3D printing can produce the channel design of 3 D stereo.Although 3D printing has many excellent
Point, but the microchannel resolution ratio that 3D printing is printed there is also an important disadvantage-is lower.
And the cross section for the microchannel that business 3D printer prints limits point of microchannel all at 100 μm or more
Resolution.Therefore, the maximum limitation of 3D printing inertia microchannel is channel resolution；And channel resolution decides that 3D printing can be beaten
The minimum channel size in channel is printed, and then influences the focusing of particle or cell, when the microchannel of 3D printing has big cross section,
It is unfavorable for the focusing of particle or cell.
[summary of the invention]
It is an object of the invention to overcome the above-mentioned prior art, a kind of 3D printing is provided and offers sinuous miniflow
The design method of the micro-fluidic chip in channel and wherein microchannel.It wriggles in the CHANNEL OPTIMIZATION micro-fluidic chip of 3D printing
The structure in channel, so that particle or tumour cell focusing effect are good under larger cross-section.
In order to achieve the above objectives, the present invention is achieved by the following scheme:
A kind of 3D printing offers the micro-fluidic chip of sinuous microchannel, including substrate, offers inside substrate micro-
Circulation road, the upper and lower surfaces of substrate are plate；One long side of substrate is first side, the long side opposite with first side
While being second side；First side offers the entrance of microchannel, and second side offers the outlet of microchannel；Miniflow is logical
Road is wave-shaped in the inside of substrate, in series by N number of arc-shaped bend, and two adjacent arc-shaped bends are in letter
" S " shape；N is the natural number more than or equal to 2；The substrate is made by 3D printing.
A further improvement of the present invention is that:
Preferably, the outlet of the entrance of microchannel and microchannel is provided with one cornerwise two of substrate respectively
End.
Preferably, the radius of curvature of arc-shaped bend is 2mm-10mm.
Preferably, the radius of curvature of arc-shaped bend is 5.9mm.
Preferably, when particle flow velocity is 480 μ L/min in microchannel, the inertia focus state of particle is best；Miniflow is logical
The particle diameter that road focuses is more than or equal to 20 μm.
A kind of 3D printing offers the design method of microchannel in the micro-fluidic chip of sinuous microchannel, including with
Lower step:
Step 1, the theoretical optimal conditions for determining that inertia focuses in microchannel；
Particle is by inertia lift F in microchannelLWith Dien power FD, when the two balance, the particle in microchannel is
Inertia focuses；It is defined as Rf, formula are as follows:
Work as RfWhen~1, particle reaches equilbrium position, is best inertia focus state；RfCalculation formula such as following formula (12) institute
Show:
In formula, r is radius of curvature, unit m；A is the diameter of particle or cell, unit m；DhIt is straight for the waterpower in channel
Diameter, unit m；ρ is fluid density, unit kg/m3；U is fluid average speed, unit m/s；μ is that the power of fluid is viscous
Degree, unit Pas；
In formula (12), a, n, ρ and μ are constant；The channel for focal diameter more than or equal to 20 μm major diameter particle and
Cell, therefore a >=20 μm；As the D of microchannelhWhen determining, RfInfluence factor be r and U, rule of thumb data, radius of curvature
Range be 2mm-10mm, flow velocity be 48-1008 μ L/min；
Step 2, it is emulated by COMSOL multiple physical field finite element analysis and determines that particle inertia focuses most in microchannel
Excellent condition；
It emulates to obtain by COMSOL multiple physical field finite element analysis, when the radius of curvature of microchannel is 5.9mm, particle
Or the focusing effect of cell is best；It emulates to obtain by COMSOL multiple physical field finite element analysis, particle or cell are logical in miniflow
When flow velocity in road is 480 μ L/min, the focusing effect of particle or cell is best；
Step 3, the optimal conditions for determining that particle inertia focuses in microchannel is emulated by focusing experimental verification；
Select particle carry out flow velocity focusing experiment, when microchannel radius of curvature be 5.9mm, by particle solution according to
Different flow velocitys are injected into microchannel, and focusing of the discovery particle when flow velocity is 480 μ L/min is most close.
Preferably, in step 1, RfCalculating process the following steps are included:
(1) inertia lift FLEmpirical formula are as follows:
In formula, ρ is fluid density, unit kg/m3；U is fluid average speed, unit m/s；A is particle diameter, single
Position is m；DhIt is the hydraulic diameter in channel, unit m；X is particle in the position by channel cross-section, and h is channel cross-section
Long-pending length, unit m；Re is Reynolds number, shown in calculation formula such as following formula (3):
In formula, μ is the dynamic viscosity of fluid, unit Pas；
(2) Dien power FDCalculation formula are as follows:
ρ is fluid density, unit kg/m in formula3；U is fluid average speed, unit m/s；A is particle or cell
Diameter, unit m；DhIt is the hydraulic diameter in channel, unit m；R is radius of curvature, unit m；
(3) R is calculatedf；
Convolution (2), formula (3), formula (7) and formula (8) can obtain:
Wherein,
δ=Dh/2r (10)
Therefore, RfCalculation formula are as follows:
In above formula, a is the diameter of particle or cell, unit m；DhIt is the hydraulic diameter in channel, unit m；R is curvature
Radius, unit m；N is constant；ρ is fluid density, unit kg/m3；U is fluid average speed, unit m/s；μ is stream
The dynamic viscosity of body, unit Pas.
Preferably, (3) step of step 1, Dien power FDCalculating process are as follows:
The calculation formula of Dien power are as follows:
FD=3 π μ UDa (4)
In formula, μ is dynamic viscosity, unit Pas；A is the diameter of particle or cell, unit m；UDIt is Dien vortex
Speed, unit m/s, shown in calculation formula such as following formula (5):
In formula, ρ is fluid fluid density, unit kg/m3, μ is dynamic viscosity, unit Pa.s；DhIt is the waterpower in channel
Diameter, unit m；DeIt is Dien number, shown in calculation formula such as following formula (6):
Convolution (4), formula (5) and formula (6), can obtain formula (7)
Preferably, when emulating radius of curvature by COMSOL multiple physical field finite element analysis, release population is set as
500, flow rate set is 500 μ L/min, and fluid density is set as 1000kg/m3, dynamic viscosity is set as 0.002Pas；
When emulating flow velocity by COMSOL multiple physical field finite element analysis, release population is set as 500, and flow rates are
48-1008 μ L/min, fluid density are set as 1000kg/m3, dynamic viscosity is set as 0.002Pas.
Preferably, it in step 3, is tested with the focusing that particle carries out flow velocity, specific steps are as follows:
(1) match particle solution；
(2) particle solution is injected into microchannel according to different flow velocitys respectively, the range of flow velocity is 48-1008 μ
L/min；
(3) fluorescent image of the lower particle different in flow rate in channel is shot；
(4) it is poly- in channel to observe lower particle different in flow rate for the fluorescent image different in flow rate shot by step (3)
It is most close to determine that particle is focused when flow velocity is 480 μ L/min for burnt situation.
Compared with prior art, the invention has the following advantages:
The micro-fluidic chip of sinuous microchannel is offered the invention discloses a kind of 3D printing；The chip is on substrate
It is provided with microchannel, microchannel is connected in series by multiple circular arcs, so that flow rates of the channel in 480-720 μ L/min
Interior to realize focusing, flow velocity is larger, and flux is high, and then realizes the focusing of particle in large scale channel；And channel design letter
Single, 3D printing is easily achieved, and is only needed a step that the printing of micro-fluidic chip and 3 D stereo microchannel can be completed, is simplified micro-
The production process of fluidic chip；Cost is lower compared with existing micro-fluidic chip manufacturing technology；And pass through 3D printing technique
The rate of recovery for the circulating tumor cell that the micro-fluidic chip of production obtains is high, being capable of high-volume volume production.
Further, the radius of curvature of arc-shaped bend is set as 2-10mm, on this basis, finds arc-shaped bend
Radius of curvature is 5.9mm, and when particle flow velocity is 480 μ L/min, the inertia focus state of particle is best.
Microchannel in the micro-fluidic chip of sinuous microchannel is offered the invention also discloses a kind of 3D printing to set
Meter method；The design method obtained by theory deduction the factor for influencing focus state of the particle in microchannel be flow velocity and
Radius of curvature after rule of thumb data determine basic scope, obtains optimal result by analogue simulation, carries out finally by experiment
Verifying, determines simulation result；The design method can be shortened the time of channel design compared with traditional design method, effective to obtain
To optimal channel structure and flow velocity.And effective solution can also be provided to design other microchannels.
[Detailed description of the invention]
Fig. 1 is a kind of solidworks of the serpentine channel micro-fluidic chip based on 3D printing provided in an embodiment of the present invention
The channel design figure of design.
Wherein 1- substrate；2- microchannel；The first side 3-；The second side 4-；5- first end；6- the second end；7-
Arc-shaped bend；
Fig. 2 a is that a kind of inertia of serpentine channel micro-fluidic chip based on 3D printing provided in an embodiment of the present invention focuses
The flow diagram of the theory analysis of particle or circulating tumor cell；
Fig. 2 b is that a kind of inertia of serpentine channel micro-fluidic chip based on 3D printing provided in an embodiment of the present invention focuses
The flow diagram of the experimental verification of particle or circulating tumor cell；
Fig. 3 a is a kind of serpentine channel micro-fluidic chip based on 3D printing provided in an embodiment of the present invention in different curvature
The box diagram of the emulation data of the particle distribution of radius lower channel outlet；
Fig. 3 b is a kind of serpentine channel micro-fluidic chip based on 3D printing provided in an embodiment of the present invention in different curvature
The thermal map of the emulation data of the particle distribution of radius lower channel outlet；
Fig. 4 a is a kind of serpentine channel micro-fluidic chip based on 3D printing provided in an embodiment of the present invention different in flow rate
The box diagram of the emulation data of the particle distribution of lower channel outlet；
Fig. 4 b is a kind of serpentine channel micro-fluidic chip based on 3D printing for providing of present example under different in flow rate
The thermal map of the emulation data of the particle distribution of channel outlet；
Fig. 5 is a kind of serpentine channel micro-fluidic chip based on 3D printing for providing of present example different in flow rate lower logical
The fluorogram that the particle of road outlet focuses；
Fig. 6 is emulation and the experiment number for a kind of serpentine channel micro-fluidic chip based on 3D printing that present example provides
According to normalization fluorescence intensity statistical chart；
Fig. 7 is a kind of serpentine channel micro-fluidic chip based on 3D printing for providing of present example in lower grain different in flow rate
The thermal map for the distribution that son goes out in channel outlet；
Fig. 8 is that a kind of inertia for serpentine channel micro-fluidic chip based on 3D printing that present example provides focuses circulation
Light field figure, fluorogram and the size distribution figure of MCF-7 and 4T1 that the experimental verification of tumour cell uses.
Fig. 9 be serpentine channel different location (entrance, centre, outlet) particle and cancer cell focus fluorescent image and
Fluorescence intensity curves are normalized, wherein flow velocity is 480 μ L/min.
Fix name " position " in all attached drawings represents position.
[specific embodiment]
The invention will be described in further detail with specific steps with reference to the accompanying drawing, and the invention discloses a kind of 3D printings
Micro-fluidic chip and its design method comprising serpentine channel.
Referring to Fig. 1, it is rectangular plate structure which, which is to select resin material, and micro-fluidic chip includes
Substrate 1 and microchannel 2, substrate 1 include two opposite long sides, respectively first side 3 and second side 4, first side 3
One end be first end 5, one end of second side 4 is the second end 6, and first end 5 and the second end 6 are right on substrate 1
The both ends of linea angulata；The entrance of microchannel 2 is offered on first end 5, and the outlet of microchannel 2 is offered on the second end 6；
Microchannel 2 is provided with the inside of substrate 1, wave-shaped, in series and two adjacent by several arc-shaped bends 7
Arc-shaped bend 7 is in alphabetical " S " shape；The radius of curvature of each arc-shaped bend 7 is 2mm-10mm, is preferably in the present embodiment
5.9mm；The optimum flow rate of the fluid flowed in microchannel 2 is 480 μ L/min；Referring to Fig. 1 it is found that microchannel is by 7 in figure
The basic unit of a " S " shape forms, and " S " shape basic unit is made of two semicircle bend pipes.
A and Fig. 2 b referring to fig. 2, the design method of the micro-fluidic chip of above-mentioned serpentine channel, wherein Fig. 2 a is theory analysis
Flow diagram；Include theory analysis major diameter microchannel inertia focus condition and COMSOL emulation find it is optimal
Radius of curvature and flow velocity；Fig. 2 b is the flow diagram of experimental verification；It is optimal that experimental verification predominantly is focused with particle
Flow velocity.
Entire design method specifically includes the following steps:
Step 1, a referring to fig. 2 is calculated and is emulated the optimal conditions that inertia focuses in determining microchannel；
(1) condition that particle inertia focuses in theory analysis microchannel；
(1-1) analyzes the stress condition of particle in microchannel；
The channel is suitable for the particle that diameter is more than or equal to 20 μm；The stress of particle includes inertia lift F in microchannelL
With Dien power FD, when the two balance, the particle in microchannel is in equilbrium position, as inertia focus state；
Wherein, inertia lift FLPhysical computing formula are as follows:
FL=Fs+Fw (1)
In formula, FsIt is shear gradient lift, FwIt is wall induced lift；
Rule of thumb formula, inertia lift FLCalculation formula such as following formula (2) shown in
ρ is fluid density, unit kg/m3；U is fluid average speed, unit m/s；A is particle diameter, and unit is
m；DhIt is the hydraulic diameter in channel, unit m；X is particle in the position by channel cross-section, and h is cross-sectional area
Length, unit m；Re is Reynolds number, shown in calculation formula such as following formula (3):
In formula, μ is the dynamic viscosity of fluid, unit Pas.
Dien power FDCalculation formula are as follows:
FD=3 π μ UDa (4)
In formula, μ is dynamic viscosity, unit Pas；A is the diameter of particle or cell, unit m；UDIt is the speed of Dien vortex
Degree, unit m/s, shown in calculation formula such as following formula (5):
In formula, ρ is fluid density, and μ is dynamic viscosity, unit m/s；DhIt is the hydraulic diameter in channel, unit m；DeIt is
Dien number, shown in calculation formula such as following formula (6):
R is radius of curvature, unit m in formula；Convolution (4)-formula (6), can obtain formula (7)
Detailed process are as follows:
Particle focuses on the condition and its influence factor of an equilbrium position in (1-2) analysis microchannel.
According to foregoing description it is found that working as inertia lift FLWith Dien power FDWhen the two balances, the particle in microchannel is in
Equilbrium position, therefore define Rf, shown in calculation formula such as following formula (8), work as R it can be seen from formula (8)fWhen~1, particle reaches flat
Weighing apparatus position.
Convolution (2) and formula (7) can obtain
Wherein,
δ=Dh/2r (10)
Therefore, RfCalculation formula are as follows:
In formula, r is the radius of curvature in channel, unit m；U is fluid average speed, unit m/s.
By RfCalculation formula is it is found that a, n, ρ and μ are constant；Since the channel is more than or equal to 20 μm for focal diameter
Major diameter particle and cell, preferably a=26 μm=2.6 × 10 in the present embodiment-5m；As the D in channelhWhen determining, radius of curvature
Two variables of r and flow velocity U will affect RfValue, in order to guarantee Rf~1, need the base of the radius of curvature and flow velocity in existing channel
Reduce flow velocity U while increasing radius of curvature r on plinth.Rule of thumb data, the range of the radius of curvature of emulation are 2mm-10mm；
The flow velocity of emulation is 48-1008 μ L/min.
From the analysis above, we can see that radius of curvature r and flow velocity U are two major parameters for influencing particle balance, therefore main below
Single factor test analogue simulation, the optimum value of both optimizations are carried out for the two parameters.
Step 2, the optimal conditions that inertia focuses is found in emulation, and the emulation total number of particles of simulation process is 500, specific to walk
It is rapid as shown in Figure 2.
(2-1) different curvature radius COMSOL multiple physical field finite element analysis emulation；
Radius of curvature is emulated by COMSOL multiple physical field finite element analysis, the parameter setting in model are as follows: release particle
Number is 500, and flow velocity is 500 μ L/min, fluid density 1000kg/m3, dynamic viscosity 0.002Pas, radius of curvature r points
It is not set as 2/3/4/5/5.5/5.8/5.85/5.9/5.95/6/6.5/7/7.5/8/9/10, unit is mm；Analogue simulation
In software, when calculation amount is at most in the channel and focusing is most close for particle, focusing effect is best.Fig. 3 a is that the present invention is real
The box diagram of the emulation data of the micro-fluidic chip particle distribution that microchannel exports under different curvature radius of example offer is provided.
As can be seen from the figure when radius of curvature is 5.9mm, particle focuses on channel center and the width focused is minimum, that is, focuses
Effect is best.Fig. 3 b is a kind of serpentine channel micro-fluidic chip based on 3D printing provided in an embodiment of the present invention in different curvature
The thermal map of the emulation data of the particle distribution of radius lower channel outlet.As can be seen from the figure radius of curvature is in 5.8mm-7.5mm
Between when, particle focuses, wherein radius of curvature be 5.9mm when, the focusing effect of particle is best.It can in conjunction with Fig. 3 a and Fig. 3 b
Calculation amount is at most in the channel and focusing is most close for particle when finding out that radius of curvature is 5.9mm, and focusing effect is best；Therefore exist
Under the setting condition, when the radius of curvature of microchannel 2 is 5.9mm, the focusing effect of particle is best.
(2-2) COMSOL multiple physical field finite element analysis emulation different in flow rate.
Flow velocity, setting model parameter are emulated by COMSOL multiple physical field finite element analysis, release population is 500, stream
Fast range is 48-1008 μ L/min, fluid density 1000kg/m3, dynamic viscosity 0.002Pas；Specific flow velocity is 48/
144/240/336/432/480/528/624/720/816/912/1008, unit is μ L/min；It is same as above, when particle is logical
When road centric quantity is at most and focusing is most close, focusing effect is best；Fig. 4 a is provided in an embodiment of the present invention micro-fluidic
The box diagram of the emulation data for the particle distribution that chip is exported in lower channel different in flow rate；As can be seen from the figure flow velocity is
Particle focuses most close when 0.1m/s (480 μ L/min), and the minimum of the width of focusing, i.e. focusing effect are best.Fig. 4 b is this hair
The thermal map of the emulation data for the particle distribution that the micro-fluidic chip that bright example provides is exported in lower channel different in flow rate；It can from figure
To find out that the particle for focusing on channel center when 480 μ L/min is most, and focus close.It can be seen that stream according to Fig. 4 a and Fig. 4 b
Calculation amount is at most in the channel and focusing is most close for particle when speed is 480 μ L/min, that is, 0.1m/s.
Step 3, experimental verification theory analysis and simulation result are focused with particle；
Whether the result obtained for verifying emulation is optimal in practical situations, passes sequentially through particle focusing experiment and tumour is thin
Born of the same parents focus and verify, and particle is red fluorescence polystyrene particle；Specifically includes the following steps:
The focusing experiment of flow velocity is carried out with particle, the radius of curvature of microchannel is 5.9mm；Specifically includes the following steps:
(1) match particle solution；
(2) particle solution in syringe is injected in microchannel by 48-1008 μ L/min flow velocity with syringe pump；
(3) fluorescent image of the particle in channel under fluorescence microscope is shot with industrial camera；
Under Fig. 5 is different in flow rate, fluorescent image of the particle in channel.As can be seen from Figure 5 240 μ L/min-720 μ
L/min occurs focusing, but the fluorescence in 480 μ L/min fluorograms is most thin, focuses best, and particle focuses on one thin
On fluorescence straight line；Therefore it is identical as the flow velocity of emulation to test the particle flow velocity obtained.
Fig. 6 is the emulation for the micro-fluidic chip that present example provides and the normalization fluorescence intensity statistics of experimental data
Figure；As can be seen from the figure only one peak value of normalization fluorescence intensity (NFI) curve of 240 μ L/min-720 μ L/min, i.e.,
Particle has focusing in channel, wherein the width of curve is minimum when 480 μ L/min, that is, focuses best.According to Fig. 6 it can be seen that returning
One changes only one peak value of fluorescence intensity curves, it was demonstrated that particle focuses a position in the channel, and normalization at this time
The FWHM of fluorescence intensity curves is minimum, it was demonstrated that the focusing of the small i.e. particle of the width of focusing of the particle in channel is close, focuses effect
Fruit is best；Therefore under conditions of other conditions determine, when flow velocity is 480 μ L/min, focusing effect is best.
Fig. 7 is the heat for the distribution that the micro-fluidic chip that present example provides goes out in lower particle different in flow rate in channel outlet
Figure；As can be seen from the figure major part particle all focuses at channel center when 480 μ L/min, and focusing effect is best.
The radius of curvature in resulting channel and flow rate result are applied into MCF-7 (human breast cancer cell) and 4T1 (mouse
Breast tumor cell) two kinds of tumour cells are focused experiment, specifically includes the following steps:
(1) MCF-7 is carried out using the micro-fluidic chip that Projet 3500HD printer produces focus experiment；
(1-1) carries out erythrocyte splitting to whole blood, removes supernatant after centrifugation.
After (1-2) is diluted remaining haemocyte, MCF-7 cell is added.
(2) 4T1 is carried out using the micro-fluidic chip that Projet 3500HD printer produces focus experiment.
Fig. 8 is that a kind of inertia for serpentine channel micro-fluidic chip based on 3D printing that present example provides focuses circulation
Light field figure, fluorogram and the size distribution figure of MCF-7 and 4T1 that the experimental verification of tumour cell uses.
Fig. 9 be serpentine channel different location (entrance, centre, outlet) particle and cancer cell focus fluorescent image and
Fluorescence intensity curves are normalized, wherein flow velocity is 480 μ L/min.The FWHM value that fluorescence intensity curves are also calculated in figure, from glimmering
The focusing effect of cell or particle is best when can be seen that 480 μ L/min in light image, fluorescence intensity curves and FWHM value,
And FWHM value 4T1 < MCF-7 < particle, 4T1 and MCF-7 cell focusing effect are better than particle focusing effect.
By focusing experimental verification, the optimum focusing radius of curvature of micro-fluidic chip is 5.9mm, micro-fluidic chip it is best
Flow velocity is 480 μ L/min.
The present invention directly prints micro-fluidic chip using 3D printing technique, only needs a step that micro-fluidic chip and three can be completed
The printing for tieing up three-dimensional microchannel, simplifies the production process of micro-fluidic chip；With existing micro-fluidic chip manufacturing technology phase
It is lower than cost；Pass through the rate of recovery height for the circulating tumor cell that the micro-fluidic chip that 3D printing technique makes obtains；
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. the micro-fluidic chip that a kind of 3D printing offers sinuous microchannel, which is characterized in that including substrate (1), substrate
(1) internal to offer microchannel (2), the upper and lower surfaces of substrate (1) are plate；One long side of substrate (1) is the first side
Side (3), opposite long side is second side (4) with first side (3)；First side (3) offers entering for microchannel (2)
Mouthful, second side (4) offer the outlet of microchannel (2)；Microchannel (2) is wave-shaped in the inside of substrate (1), by N
A arc-shaped bend (7) is in series, and two adjacent arc-shaped bends (7) are in alphabetical " S " shape；N is oneself more than or equal to 2
So number；The substrate (1) is made by 3D printing.
2. a kind of 3D printing according to claim 1 offers the micro-fluidic chip of sinuous microchannel, feature exists
In the outlet of the entrance and microchannel (2) of microchannel (2) is provided with a cornerwise both ends of substrate (1) respectively.
3. a kind of 3D printing according to claim 1 offers the micro-fluidic chip of sinuous microchannel, feature exists
In the radius of curvature of arc-shaped bend (7) is 2mm-10mm.
4. a kind of 3D printing according to claim 3 offers the micro-fluidic chip of sinuous microchannel, feature exists
In the radius of curvature of arc-shaped bend (7) is 5.9mm.
5. a kind of 3D printing according to claim 1 offers the micro-fluidic chip of sinuous microchannel, feature exists
In when the interior particle flow velocity of microchannel (2) is 480 μ L/min, the inertia focus state of particle is best；The grain that microchannel focuses
Sub- diameter is more than or equal to 20 μm.
6. a kind of 3D printing offers the design method of microchannel in the micro-fluidic chip of sinuous microchannel, feature exists
In, comprising the following steps:
Step 1, the theoretical optimal conditions for determining microchannel (2) interior inertia and focusing；
Microchannel (2) interior particle is by inertia lift FLWith Dien power FD, particle when the two balance, in microchannel (2)
For inertia focusing；It is defined as Rf, formula are as follows:
Work as RfWhen~1, particle reaches equilbrium position, is best inertia focus state；RfCalculation formula such as following formula (12) shown in:
In formula, r is radius of curvature, unit m；A is the diameter of particle or cell, unit m；DhIt is single for the hydraulic diameter in channel
Position is m；ρ is fluid density, unit kg/m3；U is fluid average speed, unit m/s；μ is the dynamic viscosity of fluid,
Unit is Pas；
In formula (12), n, ρ and μ are constant；The channel is more than or equal to 20 μm of major diameter particle and cell for focal diameter, because
This >=20 μm；As the D of microchannel (2)hWhen determining, RfInfluence factor be r and U, rule of thumb data, the model of radius of curvature
It encloses for 2mm-10mm, flow velocity is 48-1008 μ L/min；
Step 2, it is emulated by COMSOL multiple physical field finite element analysis and determines the optimal of microchannel (2) interior particle inertia focusing
Condition；
It emulates to obtain by COMSOL multiple physical field finite element analysis, when the radius of curvature of microchannel (2) is 5.9mm, particle
Or the focusing effect of cell is best；It emulates to obtain by COMSOL multiple physical field finite element analysis, particle or cell are logical in miniflow
When flow velocity in road (2) is 480 μ L/min, the focusing effect of particle or cell is best；
Step 3, the optimal conditions for determining that microchannel (2) interior particle inertia focuses is emulated by focusing experimental verification；
Select particle carry out flow velocity focusing experiment, when microchannel (2) radius of curvature be 5.9mm, by particle solution according to
Different flow velocitys is injected into microchannel (2), and focusing of the discovery particle when flow velocity is 480 μ L/min is most close.
7. 3D printing according to claim 6 offers setting for microchannel in the micro-fluidic chip of sinuous microchannel
Meter method, which is characterized in that in step 1, RfCalculating process the following steps are included:
(1) inertia lift FLEmpirical formula are as follows:
In formula, ρ is fluid density, unit kg/m3；U is fluid average speed, unit m/s；A is particle diameter, and unit is
m；DhIt is the hydraulic diameter in channel, unit m；X is particle in the position by channel cross-section, and h is cross-sectional area
Length, unit m；Re is Reynolds number, shown in calculation formula such as following formula (3):
In formula, μ is the dynamic viscosity of fluid, unit Pas；
(2) Dien power FDCalculation formula are as follows:
ρ is fluid density, unit kg/m in formula3；U is fluid average speed, unit m/s；A is the diameter of particle or cell,
Unit is m；DhIt is the hydraulic diameter in channel, unit m；R is radius of curvature, unit m；
(3) R is calculatedf；
Convolution (2), formula (3), formula (7) and formula (8) can obtain:
Wherein,
δ=Dh/2r (10)
Therefore, RfCalculation formula are as follows:
In above formula, a is the diameter of particle or cell, unit m；DhIt is the hydraulic diameter in channel, unit m；R is radius of curvature,
Unit is m；N is constant；ρ is fluid density, unit kg/m3；U is fluid average speed, unit m/s；μ is the dynamic of fluid
Power viscosity, unit Pas.
8. 3D printing according to claim 7 offers setting for microchannel in the micro-fluidic chip of sinuous microchannel
Meter method, which is characterized in that (3) step of step 1, Dien power FDCalculating process are as follows:
The calculation formula of Dien power are as follows:
FD=3 π μ UDa (4)
In formula, μ is dynamic viscosity, unit Pas；A is the diameter of particle or cell, unit m；UDIt is the speed of Dien vortex
Degree, unit m/s, shown in calculation formula such as following formula (5):
In formula, ρ is fluid density, unit kg/m3, μ is dynamic viscosity, unit Pa.s；DhIt is the hydraulic diameter in channel, it is single
Position is m；DeIt is Dien number, shown in calculation formula such as following formula (6):
Convolution (4), formula (5) and formula (6), can obtain formula (7)
9. 3D printing according to claim 6 offers setting for microchannel in the micro-fluidic chip of sinuous microchannel
Meter method, which is characterized in that when emulating radius of curvature by COMSOL multiple physical field finite element analysis, release population is set as
500, flow rate set is 500 μ L/min, and fluid density is set as 1000kg/m3, dynamic viscosity is set as 0.002Pas；
When emulating flow velocity by COMSOL multiple physical field finite element analysis, release population is set as 500, flow rates 48-
1008 μ L/min, fluid density are set as 1000kg/m3, dynamic viscosity is set as 0.002Pas.
10. 3D printing according to claim 6 offers microchannel in the micro-fluidic chip of sinuous microchannel
Design method, which is characterized in that in step 3, tested with the focusing that particle carries out flow velocity, specific steps are as follows:
(1) match particle solution；
(2) particle solution is injected into microchannel (2) according to different flow velocitys respectively, the range of flow velocity is 48-1008 μ L/
min；
(3) fluorescent image of the lower particle different in flow rate in channel is shot；
(4) fluorescent image different in flow rate shot by step (3) observes focusing feelings of the lower particle different in flow rate in channel
It is most close to determine that particle is focused when flow velocity is 480 μ L/min for condition.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102008983A (en) * | 2010-11-01 | 2011-04-13 | 武汉大学 | Microfluidic chip suitable for producing microcapsules |
| CN102411060A (en) * | 2011-12-06 | 2012-04-11 | 东南大学 | Microfluidic chip with high-aspect-ratio micro-fluidic channel and fabrication method thereof |
| CN108554467A (en) * | 2018-03-24 | 2018-09-21 | 北京工业大学 | A method of making three-dimensional microflow control chip |
-
2018
- 2018-12-07 CN CN201811496706.6A patent/CN109622076A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102008983A (en) * | 2010-11-01 | 2011-04-13 | 武汉大学 | Microfluidic chip suitable for producing microcapsules |
| CN102411060A (en) * | 2011-12-06 | 2012-04-11 | 东南大学 | Microfluidic chip with high-aspect-ratio micro-fluidic channel and fabrication method thereof |
| CN108554467A (en) * | 2018-03-24 | 2018-09-21 | 北京工业大学 | A method of making three-dimensional microflow control chip |
Non-Patent Citations (1)
| Title |
|---|
| 黄炜东: "基于惯性微流原理的微流控芯片用于血浆分离", 《科学通报》 * |
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Application publication date: 20190416 |