CN110394975A - Light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique - Google Patents
Light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique Download PDFInfo
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- CN110394975A CN110394975A CN201910608982.5A CN201910608982A CN110394975A CN 110394975 A CN110394975 A CN 110394975A CN 201910608982 A CN201910608982 A CN 201910608982A CN 110394975 A CN110394975 A CN 110394975A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/10—Processes of additive manufacturing
- B29C64/106—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material
- B29C64/118—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material using filamentary material being melted, e.g. fused deposition modelling [FDM]
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
- B29C64/10—Processes of additive manufacturing
- B29C64/106—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material
- B29C64/124—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material using layers of liquid which are selectively solidified
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y10/00—Processes of additive manufacturing
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Abstract
The present invention discloses a kind of light-cured type based on 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip.The electrophoresis path of same cross-sectional area, different materials, different cross section shape is made using the 3D printer of different printing technology, in conjunction with the liquid storage tank attachment of 3D printing, the electroosmotic flow parameter of 3D printing micro-fluidic electrophoresis chip is measured, and their electrophoretic property is investigated.Micro-fluidic electrophoresis chip production method of the present invention has fabrication cycle short, it is low in cost, the high advantage of repeatability, the defect that traditional microfluidic electrophoresis chip cost of manufacture is high, processing method is single, is difficult to realize complex configuration design is overcome, provides a kind of new technology for the production of micro-fluidic electrophoresis chip.
Description
Technical field
The present invention relates to light-cured types and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique, belong to monitoring
Technical field, the chip can be used for probing into the influence of unlike material, chip channel of different shapes to electrophoretic property.
Background technique
Microfluidic chip technology (Microfluidics) is to rely on micro-fluidic chip platform, biochemical test process
The basic operation units such as sample pretreatment, separating-purifying, detection are integrated on the chip of one piece of micro-meter scale, are automatically performed analysis
Overall process.By this technology in conjunction with electrophoresis, micro-fluidic electrophoresis chip is made, can be used for biological sample such as nucleic acid, protein, hormone
Or even the enrichment, separation and detection of cell etc., and the capillary electrophoresis in laboratory is promoted to move towards micromation, portability, low
The developing direction of cost.However the traditional manufacturing technique of micro-fluidic chip is such as: silicon/polymer surfaces micro Process, soft print, pressure
The process such as print, injection moulding, laser ablation require to complete in ultra-clean chamber, and process is various, complex procedures, occupy a large amount of
Space, it is complicated for operation, need exper ienced design and processing staff and expensive.So being badly in need of a kind of revolutionary character at this stage
Technology replace traditional micro-fluidic manufacture craft.
In recent years, 3D printing technique is because its is easy to operate, controllability is strong, the advantages such as at low cost cause micro-fluidic field
Extensive concern.3D printing technique is to carry out rapid shaping using lamination techniques.It and the basic phase of common plane print job principle
Together, only replace the ink in normal printer with consumptive materials such as liquid or powder, by Slice Software 3D model cutting at
Plane layer by layer gets up consumptive material layer upon layer after printer identification, and the blueprint on computer is finally become in kind.Nowadays, 3D
Printer has been able to manufacture micron order even nanoscale characteristic size, it is sufficient to reach most of laboratory for micro-fluidic core
The required precision of piece.Meanwhile the micro-fluidic chip of 3D printing can allow researcher to explore fluid motion in real three-dimensional structure
Secret.Importantly, this technology of 3D printing greatly reduces the period of micro-fluidic chip " design-manufacture-test ", energy
Fast implement the idea of researcher.
The present invention discloses light-cured type based on 3D printing technique and fusion sediment type electrophoresis chip, utilization are different types of
3D printer make same cross-sectional area, different materials, different cross section shape electrophoresis path, and to their electrophoretic property into
Row comparison.
Summary of the invention
1. the purpose of the present invention is establish light-cured type and the micro-fluidic electrophoresis core of fusion sediment type based on 3D printing technique
Piece and its application.
The object of the present invention is achieved like this, utilizes the micro-fluidic electricity of the 3D printer production of two kinds of different printing technology
Swimming chip, two kinds of printers are respectively light-cured type (SLA) 3D printer and fusion sediment type (FDM) 3D printer.
The micro-fluidic electrophoresis chip is by including that modelling, software slice and micro-fluidic electrophoresis chip print three mistakes
Journey production, wherein SLA type micro-fluidic electrophoresis chip manufacturing process is as follows: (1) modeling: utilizing 123D Design software building
Chip three-dimensional structure, design microchannel plate are 5 having a size of the mm × 2 of 70 mm × 20 mm, two identical liquid storage tank diameter d
Mm, height h are 2 mm, and micro-tunnel length is 57 mm, and cross-sectional area is 100 μm2, each chip includes two identical electrophoresis
Channel；The bottom plate having a size of the mm × 2 of 70 mm × 20 mm is constructed, surface guarantees smooth；(2) it is sliced: upper step is virtually counted
The output of word 3D model is stl format, chip model is carried out hierarchy slicing, design parameter with Preform software are as follows: photocuring material
Material: clear V4(FLGPCL04)；Print thickness: 0.025 mm；Tilt angle: X, Y, Z axis respectively tilts 45 degree；Whether support:
Addition support；The two-dimensional silhouette model that Preform software is sliced to obtain each layer cross section is uploaded to light-cured type with .stl format
3D printer；(3) it prints: opening printer, resin cartridge is opened in leveling, and resin automatically flows into resin storage tank, selects the printing needed
Model starts to print after resin storage tank temperature is raised to 31 degree；(4) post-process: printing is completed, and is opened printer lid, is removed structure
Jian Pingtai prizes lower chip, puts it into cleaning box；Microchannel plate and bottom plate are first immersed in the cleaning box of anhydrous isopropyl alcohol, vibration
Washing 20-30 min is shaken, is then placed in the cleaning box of dehydrated alcohol and impregnates 5 min to wash away uncured resin；(5) it is hot pressed into
Type: taking out microchannel plate and bottom plate removes backing material, and room temperature dries, and heat gun is transferred to 210 degree, is evenly heated 3 min of bottom plate,
The seamless lid of bottom plate and microchannel plate is closed rapidly, is fitted into polyethylene terephthalate (PET) set, then moves to brave bench vice
Jaw, it is with the powerful pressure of brave bench vice that two plates are seamless applying together；Then, electrophoresis chip hot pressing being bonded, is placed in
1 h is irradiated under 405 nm ultraviolet lamps；Finally, bonding the heightening device of 3D printing at liquid storage tank, increased heights are 5 mm, to
Expand liquid storage tankage, eventually forms SLA type electrophoresis chip；FDM type electrophoresis chip manufacturing process is as follows: (1) modeling: utilizing
123D Design software building chip three-dimensional structure, design channel shape are the electrophoresis path of regular quadrangle, and cross-sectional area is
100 μm2, long 57 mm, channel upper layer and underlayer thickness are respectively 2 mm, accelerate heat with this and distribute；Liquid storage tank is that diameter is
There is the cylinder of cavity in 5 mm, a height of 7 mm, enough sample volumes are guaranteed with this；(2) it is sliced: by above-mentioned virtual digit mould
Type output is .stl format, is placed in JGcreate Slice Software, and a height of 0.1 mm of layer is arranged, and print density is 100 %, spray
210 DEG C of temperature of head, 50 DEG C of hott bed temperature, print speed is 30 mm/s, and movement speed is 70 mm/s；It, will after being provided with
This document saves as .gcode format, is uploaded in FDM type 3D printer with USB flash disk；(3) it prints: adjustment printer podium level, really
The distance between spray head and platform are the thickness of an A4 paper when protecting initialization；The poly- cream of high quality white is imported after preheating 5 min
Sour (PLA) consumptive material, selects above-mentioned file to start to print；(4) it post-processes: printed model is put into ultrasound immersion in ultrapure water
2 times, 15 min, subsequent cold wind dry up every time；It is persistently slowly introducing 40 % chloroforms in the molded passage of drying, keeps microchannel
1 min is impregnated in the chloroformic solution of flowing.
The light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique, which is characterized in that
Construct a dismountable electrophoresis splicing apparatus by following step, electrophoresis splicing apparatus anabolic process is as follows: (1) electrophoresis splices
The design of device: the device is the inverse u shape structure being made of three faces, and there are four the circular holes that diameter is 5 mm for putting at top
Set electrode；(2) it the assembling of chip: by the electrophoresis chip of 3D printing in the inverted " u " type groove of electrophoresis splicing apparatus, then places
On ice chest, to reduce the influence of Joule heat；(3) connection of electrode: electrode is quasi- using the top circular hole of electrophoresis splicing apparatus
It is really inserted into the liquid storage tank of chip, and electrode is connect by conducting wire with high voltage power supply.
The light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique, which is characterized in that
SLA type 3D printer is transparent photosensitive resin clear V4(FLGPCL04 using material), heightening device uses the 3D printing of FDM type
Machine printing, 3D printing material are transparent rubber；FDM type 3D printer is white PLA using material.
The light-cured type and fusion sediment type electrophoresis chip based on 3D printing technique, which is characterized in that with identical
Cross-sectional area, is selected from equilateral triangle, regular quadrangle, 3/4 positive icosagon or 3/4 circular varying cross-section shape at equal length
The SLA type electrophoresis chip of shape is research object, and with phosphate buffer solution (PBS) for buffer, electroendosmotic mobility increases with pH
It is big and gradually become smaller, and the slope of curve of the regular quadrangle channel at pH=5 ~ 7 is maximum.
The light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique, which is characterized in that
It is slow with phosphate buffer solution (PBS) using SLA type and FDM type regular quadrangle channel microfluidic electrophoresis chip as research object
The electroendosmotic mobility of fliud flushing, SLA type micro-fluidic electrophoresis chip increases with pH and is reduced, and optimal acidity is pH=5；FDM type miniflow
The electroendosmotic mobility for controlling electrophoresis chip increases with pH and is increased, and optimal acidity is pH=9.
The application of the above-mentioned light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique of the present invention,
It is characterized in that, being applied to the monitoring of electroosmotic flow.
2. specifically, of the present invention making equilateral triangle, regular quadrangle, 3/4 just using SLA type 3D printer
The electrophoresis path of icosagon and 3/4 circular shape, sequentially includes the following steps:
(1) it models
Using 123D Design software building chip three-dimensional structure, microchannel plate is designed having a size of mm × 2 70 mm × 20
Mm, two identical liquid storage tank diameter d=5 mm, height h=2 mm, 57 mm of micro-tunnel length, cross-sectional area are 100 μ
m2, each chip includes two identical electrophoresis paths；The bottom plate having a size of the mm × 2 of 70 mm × 20 mm is constructed, surface is not
Design configuration guarantees smooth.
(2) it is sliced
It by virtual 3d model in (1), exports as .stl format, chip 3D model is subjected to hierarchy slicing with Preform software, select
Selecting material is clear V4(FLGPCL04)；Setting printing 0.025 mm of thickness；Adjust chip and print angle: X, Y, Z axis is respectively inclined
Oblique 45 degree.Then addition support must ensure that support does not fall in microchannel plate and backplate surface.Finally, Preform software is cut automatically
Piece obtains the two-dimensional silhouette model of each layer cross section.Computer is connected with photocuring 3D printer, it then will be by slice
.stl format tomograph imports Form2 3D printer.
(3) it prints
Printer is opened, resin cartridge is opened in leveling, and resin automatically flows into resin storage tank, the printer model needed is selected, to resin storage tank
After temperature is raised to 31 degree, start to print.
(4) it post-processes
Printing is completed, and printer lid is opened, and removes construction platform, is prized lower chip, is put it into cleaning box.Chip is first immersed in
In the cleaning box of anhydrous isopropyl alcohol, shaking washing 20-30 min, be then placed in the cleaning box of dehydrated alcohol impregnate 5 min with
Wash away uncured resin.
(5) hot-forming
It takes out chip and removes its backing material, room temperature dries 3-5 min, and heat gun is transferred to 210 degree of low grades, is evenly heated bottom plate 3
Then min rapidly closes the seamless lid of bottom plate and microchannel plate, be fitted into special PET set, then move to brave bench vice jaw, use
The powerful pressure of brave bench vice is seamless applying together by two plates.Then, electrophoresis chip hot pressing being bonded, it is ultraviolet to be placed in 405 nm
1 h is shone under lamp.Finally, bonding the heightening device of 3D printing at liquid storage tank, increased heights are 5 mm, to expand liquid storage pool capacity
Amount.
It sequentially include such as 3. of the present invention make regular quadrangle micro-fluidic electrophoresis chip using FDM type 3D printer
Lower step:
(1) it models
Using 123D Design software building chip three-dimensional structure, it is logical to design the micro-fluidic electrophoresis that channel shape is regular quadrangle
Road, cross-sectional area are 100 μm2, long 57 mm, channel upper layer and underlayer thickness are respectively 2 mm, accelerate heat with this and distribute.
Liquid storage tank is that diameter is 5 mm, and the cylinder of a height of 7 mm guarantees enough sample volumes with this.
(2) it is sliced
Above-mentioned virtual digital model is exported as .stl format, is placed in JGcreate Slice Software, layer a height of 0.1 is set
Mm, print density are 100 %, and 210 DEG C of nozzle temperature, 50 DEG C of hott bed temperature, print speed is 30 mm/s, and movement speed is
70 mm/s.After being provided with, this document is saved as into .gcode format, is uploaded in FDM type 3D printer with USB flash disk.
(3) it prints
Adjust printer podium level, it is ensured that the distance between spray head and platform are the thickness of an A4 paper, preheating 5 when initialization
High quality white polylactic acid (PLA) consumptive material is imported after min, and above-mentioned file is selected to start to print.
(4) it post-processes
Printed model is put into ultrasound in ultrapure water to impregnate 2 times, 15 min, subsequent cold wind dry up every time.In the mould of drying
It is persistently slowly introducing 40 % chloroforms in type channel, microchannel is kept to impregnate 1 min in the chloroformic solution of flowing.
4. the light-cured type and fusion sediment type micro-fluidic electrophoresis chip monitoring of the present invention based on 3D printing technique
The process of electroosmotic flow, sequentially includes the following steps:
(1) electroosmotic flow monitoring device is built
Electrophoresis chip and detecting electrode are fitted together by the electrophoresis splicing apparatus of 3D printing.The electrophoresis splicing apparatus is by three
The inverse u shape structure that a face is constituted, there are four the circular holes that diameter is 5 mm for placing electrode at top.The device is whole in addition to that can make
The operation of a electroosmotic flow monitoring process is more convenient, moreover it is possible to will test the relative position of electrode and micro-fluidic electrophoresis chip liquid storage tank
It is fixed, reduce experimental error.Device is placed on ice chest in experimentation, for reducing the joule generated in electrophoresis process
Heat.
(2) monitoring of electroosmotic flow
First chip is placed in deionized water before detection and is cleaned by ultrasonic 10 min, deionized water is then added in microchannel, is used
Vacuum pump depletion cleans 2 times, then plus solution to be measured cleaning 2 times.Using vacuum pump, make to be full of the 3.0 of different acidity in microchannel
mM PBS.Positive and negative electrode is inserted into microchannel both ends respectively, opens high voltage power supply, bonus point ionization voltage, after electric current is stablized, pause is electric
Swimming, observes and records current-vs-time figure.Remaining buffering in the liquid storage tank for placing positive electrode is then sucked out with injector for medical purpose
Liquid is added Micro Phases with 1.8 mM PBS of acidity, applies separation voltage again, and current value can be observed and be gradually reduced finally surely
It is fixed, current-vs-time figure is observed and recorded, the time t for dropping to low current from high current is read.According to electrophoretic mobility calculation formula
μEOF= L2/ (tV) calculates electrophoretic mobility μEOF。
The invention has the advantages that
This experiment makes micro-fluidic electrophoresis chip using 3D printing technique, and speed is fast, and price is low, and repeatability is high, and has printed phase
It is compared with cross-sectional area electrophoresis path of different shapes, provides a kind of new technology for the production of electrophoresis chip.
Detailed description of the invention
Fig. 1 is that the present invention is based on the dresses of the light-cured type of 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip device
With schematic diagram.(in figure: 1:3D printing shaping electrophoresis chip；2: microchannel plate；3: bottom plate；4: microchannel plate-bottom of hot pressing synthesis
Plate；5: heightening device；6:SLA type electrophoresis chip；7:FDM type electrophoresis chip；8:FDM type 3D printer；9: electrophoresis splicing apparatus；
10: ice chest；11: positive electrode；12: negative electrode；13: high-voltage chip electrophoresis apparatus；14: computer).
Fig. 2 is that the present invention is based on the light-cured types of 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip channel cross-section
Micro- in kind relatively scheme.
The present invention is based on printed materials in the light-cured type of 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip by Fig. 3
The x-ray photoelectron spectroscopy of photosensitive resin sweeps figure (3a), C 1S figure (3b), N 1S figure (3c) entirely.
Fig. 4 is that the present invention is based on SLA types in the light-cured type of 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip
Different cross section shape electrophoresis chip electroosmotic flow monitoring figure.
Fig. 5 is SLA in the light-cured type and fusion sediment type micro-fluidic electrophoresis chip of the invention based on 3D printing technique
The electroosmotic flow of type and FDM type regular quadrangle channel electrophoresis chip monitoring figure.
Specific embodiment
In order to be more clear the technical problem to be solved in the present invention, technical solution and using effect, below in conjunction with implementation
Example and attached drawing, the present invention will be described in further detail.
As shown in Figure 1, the light-cured type and the micro-fluidic electrophoresis of fusion sediment type of the present invention based on 3D printing technique
Chip apparatus figure: the 3D printing molding electrophoresis chip 1 that 3D printing is completed is put in inside the electrophoresis splicing apparatus 9 of 3D printing, together
When, positive electrode 11, negative electrode 12 are also inserted into the aperture above electrophoresis splicing apparatus, and they are gently placed on ice chest 10 it
On.Whole device is immobilized and is conducive to improve the stability of test.Then, positive electrode 11, the electricity on 12 electrode of negative electrode
Line is connected in high-voltage chip electrophoresis apparatus 13, and last electrophoresis apparatus connects computer 14 by USB line.
Above-mentioned electrophoresis splicing apparatus makes in the steps below, and electrophoresis splicing apparatus anabolic process is as follows: (1) electrophoresis splices
The design of device: the device is the inverse u shape structure being made of three faces, and there are four the circular holes that diameter is 5 mm for putting at top
Set electrode；(2) it the assembling of chip: by the electrophoresis chip of 3D printing in the inverted " u " type groove of electrophoresis splicing apparatus, then places
On ice chest, to reduce the influence of Joule heat；(3) connection of electrode: electrode is quasi- using the top circular hole of electrophoresis splicing apparatus
It is really inserted into the liquid storage tank of chip, and electrode is connect by conducting wire with high voltage power supply.
Embodiment 1:
One) electrophoresis of SLA type 3D printer production equilateral triangle, regular quadrangle, 3/4 positive icosagon and 3/4 circular shape is logical
Road sequentially includes the following steps:
(1) it models
Using 123D Design software building chip three-dimensional structure, design microchannel plate size (appended drawing reference 2 in Fig. 1), it is high
The mm of h=2,57 mm of micro-tunnel length are spent, cross-sectional area is 100 μm2, each microchannel plate includes that two identical electrophoresis are logical
Road 2-1；The bottom plate (appended drawing reference 3 in Fig. 1) having a size of the mm × 2 of 70 mm × 20 mm is constructed, surface not design configuration is protected
It demonstrate,proves smooth；Two identical heightening device (appended drawing reference 5 in Fig. 1) diameter d=5 mm.
(2) it is sliced
By virtual 3d model in step (1), exports as .stl format, chip 3D model is subjected to layering with Preform software and is cut
Piece selects material for clear V4(FLGPCL04), it is commercial product；Setting printing 0.025 mm of thickness；Adjust chip printing
Angle: X, Y, Z axis respectively tilts 45 degree.Then addition support must ensure that support does not fall in microchannel plate and backplate surface.Finally,
Preform software auto slice obtains the two-dimensional silhouette model of each layer cross section.Computer is connected with photocuring 3D printer, so
Afterwards Form2 3D printer will be imported by the .stl format tomograph of slice.
(3) it prints
Printer is opened, resin cartridge is opened in leveling, and resin automatically flows into resin storage tank, the printer model needed is selected, to resin storage tank
After temperature is raised to 31 degree, start to print.
(4) it post-processes
Printing is completed, and printer lid is opened, and removes construction platform, prizes lower chip (microchannel plate and bottom plate), is put it into clear
Wash box.Chip (microchannel plate and bottom plate) is first immersed in the cleaning box of anhydrous isopropyl alcohol, shaking washing 20-30 min, then
It is put into the cleaning box of dehydrated alcohol and impregnates 5 min to wash away uncured resin.
(5) hot-forming
Take out chip (microchannel plate and bottom plate) and remove its backing material, room temperature dries 3-5 min, heat gun be transferred to 210 degree it is low
Shelves, are evenly heated 3 min of bottom plate, and then the seamless lid of bottom plate and microchannel plate is closed rapidly, are fitted into special PET set, then
Brave bench vice jaw is moved to, it is with the powerful pressure of brave bench vice that two plates are seamless applying together.Then, hot pressing hot pressing being bonded
At microchannel plate-bottom plate 4, be placed under 405 nm ultraviolet lamps according to 1 h.Finally, being glued at round liquid storage tank on microchannel plate
The heightening device 5 of 3D printing is closed, increased heights are 5 mm, and heightening device 5 is the cylinder that diameter is 5 mm, to expand liquid storage tank
Capacity；It eventually forms SLA type electrophoresis chip 6(and sees Fig. 1).
This experiment observes microchannel cross-section using inverted microscope.As shown in Fig. 2, a in figure is positive triangle
B in shape microchannel cross-section, figure is regular quadrangle microchannel cross-section, the c in figure is that 3/4 positive icosagon microchannel is transversal
D in face and figure is 3/4 round microchannel cross-section；Wherein 3/4 positive icosagon and 3/4 circular profile are similar, because
Under minute yardstick, 3/4 positive icosagon leveled off to 3/4 circle.Therefore it can determine whether that 2 3D printer of Form can print cross section
Product equal equilateral triangle, regular quadrangle, 3/4 positive icosagon and 3/4 circular microchannel.
Two) FDM type 3D printer makes regular quadrangle electrophoresis path, sequentially includes the following steps:
(1) it models
Using 123D Design software building chip three-dimensional structure, the electrophoresis path that channel shape is regular quadrangle is designed, it is transversal
Area is 100 μm2, long 57 mm, channel upper layer and underlayer thickness are 2 mm, accelerate heat with this and distribute；Liquid storage tank is straight
Diameter is 5 mm, and the cylinder of a height of 7 mm guarantees enough sample volumes with this.
(2) it is sliced
Above-mentioned virtual digital model is exported as .stl format, is placed in JGcreate Slice Software, layer a height of 0.1 is set
Mm, print density are 100 %, and 210 DEG C of nozzle temperature, 50 DEG C of hott bed temperature, print speed is 30 mm/s, and movement speed is
70 mm/s；After being provided with, this document is saved as into .gcode format, is uploaded in FDM type 3D printer with USB flash disk.
(3) it prints
Adjust printer podium level, it is ensured that the distance between spray head and platform are the thickness of an A4 paper when initialization；Preheating 5
High quality white polylactic acid (PLA) consumptive material is imported after min, and above-mentioned file is selected to start to print.
(4) it post-processes:
Printed model is put into ultrasound in ultrapure water to impregnate 2 times, 15 min, subsequent cold wind dry up every time.In the mould of drying
It is persistently slowly introducing 40wt% chloroform in type channel, microchannel is kept to impregnate 1 min in the chloroformic solution of flowing, forms FDM type
Electrophoresis chip 7(is shown in Fig. 1).
Embodiment 2:
In order to verify the specific chemical component in microchannel, this experiment is characterized using x-ray photoelectron spectroscopy (XPS), as a result
As shown in Figure 3.It sweeps figure entirely by a in Fig. 3 to know, photosensitive resin mainly contains C, N, O element, and wherein C, O constituent content are relatively abundant.By
The C 1S figure of b in Fig. 3 knows that 284.80 eV of peak position is C=C, and 285.62 eV of peak position is C-N, and peak position 286.68 is C-
O, it is O-C=O that peak position, which is set to 289.19 eV, meets the chemical bond feature of methacrylate.Known by the N 1S figure of the c in Fig. 3,
N element only has a kind of valence state, and peak position is the structure of secondary nitrogen in 400.01 eV.In conjunction with the information that producer provides, the master of the material
Wanting ingredient is methacrylate, after nitrogenous photoinitiator is added, forms the polymer containing parahelium structure, has in conjunction with hydrogen
The potentiality of ion.
Embodiment 3:
Surpass firstly, chip made from embodiment 1 (SLA type electrophoresis chip 6 or FDM type electrophoresis chip 7) is placed in deionized water
Sound cleans 10 min.Then, deionized water is added in microchannel, is cleaned 2 times using vacuum pump, then plus solution to be measured cleaning 2 times.
Then, clean chip is placed on ice chest.Made in microchannel with vacuum pump full of micro different acidity, 3.0 mM of concentration
PBS.Positive and negative electrode is inserted into microchannel both ends, opening chip electrophoresis apparatus, and bonus point ionization voltage U suspends electrophoresis after stabling current,
Observe and record current-vs-time figure.Residual buffer liquid in the liquid storage tank for placing positive electrode is sucked out with injector for medical purpose, is added micro
1.8 mM PBS of identical acidity.Apply separation voltage U at this time, current value can be observed and be gradually reduced last stabilization, observation is simultaneously
Record current-time diagram reads the time t for dropping to low current from high current.According to electrophoretic mobility formula μEOF=L2/ (tV)
Calculate electrophoretic mobility μEOF。
As shown in Figure 4, SLA type micro-fluidic electrophoresis chip microchannel of various shapes (equilateral triangle: n=3；Regular quadrangle: n
=4；3/4 positive icosagon n=20；3/4 is round: n=∞) electric osmose mobility is all reduced as pH is increased, this illustrates the acid of solution
Degree, which reduces, causes the effective charge density of microchannel surface to reduce, and according to the forming process of electric double layer, microchannel surface is positively charged
Lotus.Speculated according to XPS map, it should be the nitrogenous compound binding hydrogen ions formation nominal price compound of material itself, make micro- logical
Road surface is positively charged.
After pH value of solution=7, electroendosmotic mobility decline is gradually gentle, illustrates that microchannel surface effective charge is a large amount of
Inhibit.And before pH value of solution=7, the electrophoretic mobility in regular quadrangle channel is maximum, next triangular duct that is positive is finally circle
Shape channel and positive icosagon channel.Known by Fig. 4, the cross section characteristic profile substantially phase of positive icosagon channel and circular channel
When, therefore the curve of the two is also roughly the same.And the slope of a curve of n=4 is maximum, and electric osmose mobility is maximum in pH=5, illustrates positive four
Side shape channel is most sensitive to solution charge variation, is more suitable for doing the channel of electrophoretic analysis, optimal acidity is pH=5.
Embodiment 4:
Different printers make different materials in light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique
Expect that the step of micro-fluidic electrophoresis chip is monitored for electroosmotic flow is as follows:
100 μm of sectional area are made to SLA type printer and transparent photosensitive resin using identical method and condition in embodiment 22
Square electrophoresis path and FDM type printer and PLA make 100 μm of sectional area2The electroosmotic flow of square electrophoresis path carries out pair
Than such as Fig. 5.
As shown in Figure 5, FDM type printer and PLA production micro-fluidic electrophoresis chip electroendosmotic mobility value with pH increase and
Increase.Illustrate to reduce with buffer solution system acidity, the charge density on channel surface increases, therefore according to electric double layer principle, leads to
Road surface should be distributed negative electrical charge.And FDM type beats curve linear that micro-fluidic print chip pH change with electric osmose mobility more preferably, and it is electric
Infiltration mobility value is lower, and error bar is smaller.This may be because PLA chemical property it is more stable, while FDM printing be once at
Type, therefore the otherness of each chip chamber is smaller, it is as a result more stable.
Claims (7)
1. a kind of light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique, which is characterized in that utilize
The micro-fluidic electrophoresis chip of the 3D printer production of two kinds of different printing technology, two kinds of printers are respectively light-cured type (SLA)
3D printer and fusion sediment type (FDM) 3D printer.
2. the light-cured type based on 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip as described in claim 1,
It is characterized in that, the micro-fluidic electrophoresis chip is by including that modelling, software slice and micro-fluidic electrophoresis chip print three
Process production, wherein SLA type micro-fluidic electrophoresis chip manufacturing process is as follows: (1) modeling: utilizing 123D Design software structure
Chip three-dimensional structure is built, designs microchannel plate having a size of the mm × 2 of 70 mm × 20 mm, two identical liquid storage tank diameter d
It is 2 mm for 5 mm, height h, micro-tunnel length is 57 mm, and cross-sectional area is 100 μm2, each chip includes two identical
Electrophoresis path；The bottom plate having a size of the mm × 2 of 70 mm × 20 mm is constructed, surface guarantees smooth；(2) it is sliced: upper step is empty
Quasi- number 3D model output is stl format, chip model is carried out hierarchy slicing, design parameter with Preform software are as follows: light is solid
Change material: clear V4(FLGPCL04)；Print thickness: 0.025 mm；Tilt angle: X, Y, Z axis respectively tilts 45 degree；Whether prop up
Support: addition support；The two-dimensional silhouette model that Preform software is sliced to obtain each layer cross section is uploaded to photocuring with .stl format
Type 3D printer；(3) it prints: opening printer, resin cartridge is opened in leveling, and resin automatically flows into resin storage tank, selects what is needed to beat
Stamp type starts to print after resin storage tank temperature is raised to 31 degree；(4) post-process: printing is completed, and is opened printer lid, is removed
Construction platform prizes lower chip, puts it into cleaning box；Microchannel plate and bottom plate are first immersed in the cleaning box of anhydrous isopropyl alcohol,
Shaking washing 20-30 min, is then placed in the cleaning box of dehydrated alcohol and impregnates 5 min to wash away uncured resin；(5) hot pressing
Molding: taking out microchannel plate and bottom plate removes backing material, and room temperature dries, and heat gun is transferred to 210 degree, is evenly heated bottom plate 3
Min rapidly closes the seamless lid of bottom plate and microchannel plate, is fitted into polyethylene terephthalate (PET) set, then moves to tiger
Bench vice jaw, it is with the powerful pressure of brave bench vice that two plates are seamless applying together；Then, electrophoresis chip hot pressing being bonded, puts
It is placed under 405 nm ultraviolet lamps and irradiates 1 h；Finally, bonding the heightening device of 3D printing at liquid storage tank, increased heights are 5 mm,
To expand liquid storage tankage, SLA type electrophoresis chip is eventually formed；FDM type electrophoresis chip manufacturing process is as follows: (1) modeling: benefit
With 123D Design software building chip three-dimensional structure, the electrophoresis path that channel shape is regular quadrangle is designed, cross-sectional area is
100 μm2, long 57 mm, channel upper layer and underlayer thickness are respectively 2 mm, accelerate heat with this and distribute；Liquid storage tank is that diameter is
There is the cylinder of cavity in 5 mm, a height of 7 mm, enough sample volumes are guaranteed with this；(2) it is sliced: by above-mentioned virtual digit mould
Type output is .stl format, is placed in JGcreate Slice Software, and a height of 0.1 mm of layer is arranged, and print density is 100 %, spray
210 DEG C of temperature of head, 50 DEG C of hott bed temperature, print speed is 30 mm/s, and movement speed is 70 mm/s；It, will after being provided with
This document saves as .gcode format, is uploaded in FDM type 3D printer with USB flash disk；(3) it prints: adjustment printer podium level, really
The distance between spray head and platform are the thickness of an A4 paper when protecting initialization；The poly- cream of high quality white is imported after preheating 5 min
Sour (PLA) consumptive material, selects above-mentioned file to start to print；(4) it post-processes: printed model is put into ultrasound immersion in ultrapure water
2 times, 15 min, subsequent cold wind dry up every time；It is persistently slowly introducing 40 % chloroforms in the molded passage of drying, keeps microchannel
1 min is impregnated in the chloroformic solution of flowing.
3. the light-cured type based on 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip as described in claim 1,
It is characterized in that, a dismountable electrophoresis splicing apparatus is constructed by following step, electrophoresis splicing apparatus anabolic process is as follows:
(1) design of electrophoresis splicing apparatus: the device is the inverse u shape structure being made of three faces, and top is 5 mm there are four diameter
Circular hole for placing electrode；(2) electrophoresis chip of 3D printing the assembling of chip: is embedded in the inverse u shape of electrophoresis splicing apparatus
It in slot, is then placed on ice chest, to reduce the influence of Joule heat；(3) connection of electrode: electrode utilizes electrophoresis splicing apparatus
Top circular hole be properly inserted in the liquid storage tank of chip, and electrode is connect by conducting wire with high voltage power supply.
4. the light-cured type based on 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip as described in claim 1,
It is characterized in that, SLA type 3D printer is transparent photosensitive resin clear V4(FLGPCL04 using material), heightening device uses
The printing of FDM type 3D printer, 3D printing material are transparent rubber；FDM type 3D printer is white PLA using material.
5. the light-cured type based on 3D printing technique and fusion sediment type electrophoresis chip, feature exist as described in claim 1
In with same cross-sectional area, equal length, selected from equilateral triangle, regular quadrangle, 3/4 positive icosagon or 3/4 circular difference
The SLA type electrophoresis chip of cross-sectional shape is research object, with phosphate buffer solution (PBS) for buffer, electric osmose trickling
Degree increases with pH and is gradually become smaller, and the slope of curve of the regular quadrangle channel at pH=5 ~ 7 is maximum.
6. the light-cured type based on 3D printing technique and fusion sediment type micro-fluidic electrophoresis chip as described in claim 1,
It is characterized in that, using SLA type and FDM type regular quadrangle channel microfluidic electrophoresis chip as research object, with phosphate buffer solution
It (PBS) is buffer, the electroendosmotic mobility of SLA type micro-fluidic electrophoresis chip increases with pH and reduced, and optimal acidity is pH=5；
The electroendosmotic mobility of FDM type micro-fluidic electrophoresis chip increases with pH and is increased, and optimal acidity is pH=9.
7. any light-cured type and fusion sediment type micro-fluidic electrophoresis chip based on 3D printing technique of claim 1-6
Application, which is characterized in that the monitoring applied to electroosmotic flow.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111589477A (en) * | 2020-05-28 | 2020-08-28 | 韶关学院 | Micro-channel device processing technology |
| CN111974471A (en) * | 2020-06-23 | 2020-11-24 | 江苏敦超电子科技有限公司 | 3D printing-based microfluidic electrical detection chip processing method |
| CN114062470A (en) * | 2021-09-29 | 2022-02-18 | 福建医科大学 | 3D printing array type BP-LED-E sensing device for hydrogen peroxide detection |
| CN114055772A (en) * | 2021-09-29 | 2022-02-18 | 福建医科大学 | Manufacturing method of 3D printing assembling paper-based micro-fluidic chip |
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| CN103285950A (en) * | 2013-05-27 | 2013-09-11 | 苏州扬清芯片科技有限公司 | Preparation method for polymer micro-fluidic chip |
| CN107519960A (en) * | 2017-09-06 | 2017-12-29 | 清华大学 | Microchannel and preparation method thereof |
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| CN103285950A (en) * | 2013-05-27 | 2013-09-11 | 苏州扬清芯片科技有限公司 | Preparation method for polymer micro-fluidic chip |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111589477A (en) * | 2020-05-28 | 2020-08-28 | 韶关学院 | Micro-channel device processing technology |
| CN111589477B (en) * | 2020-05-28 | 2022-04-15 | 韶关学院 | Micro-channel device processing technology |
| CN111974471A (en) * | 2020-06-23 | 2020-11-24 | 江苏敦超电子科技有限公司 | 3D printing-based microfluidic electrical detection chip processing method |
| CN114062470A (en) * | 2021-09-29 | 2022-02-18 | 福建医科大学 | 3D printing array type BP-LED-E sensing device for hydrogen peroxide detection |
| CN114055772A (en) * | 2021-09-29 | 2022-02-18 | 福建医科大学 | Manufacturing method of 3D printing assembling paper-based micro-fluidic chip |
| CN114062470B (en) * | 2021-09-29 | 2023-06-13 | 福建医科大学 | 3D printing array type BP-LED-E sensing device for hydrogen peroxide detection |
| CN114055772B (en) * | 2021-09-29 | 2023-10-20 | 福建医科大学 | Manufacturing method of 3D printing assembled paper-based microfluidic chip |
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