CN114107025A - Fluid driving method based on algae cells - Google Patents
Fluid driving method based on algae cells Download PDFInfo
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- CN114107025A CN114107025A CN202111439893.6A CN202111439893A CN114107025A CN 114107025 A CN114107025 A CN 114107025A CN 202111439893 A CN202111439893 A CN 202111439893A CN 114107025 A CN114107025 A CN 114107025A
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- 239000012530 fluid Substances 0.000 title claims abstract description 186
- 241000195493 Cryptophyta Species 0.000 title claims abstract description 143
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000006073 displacement reaction Methods 0.000 claims abstract description 10
- 210000004027 cell Anatomy 0.000 claims description 145
- 239000011521 glass Substances 0.000 claims description 75
- 229910021417 amorphous silicon Inorganic materials 0.000 claims description 33
- 210000003495 flagella Anatomy 0.000 claims description 15
- 238000004720 dielectrophoresis Methods 0.000 claims description 13
- 239000008367 deionised water Substances 0.000 claims description 12
- 229910021641 deionized water Inorganic materials 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 230000007246 mechanism Effects 0.000 claims description 10
- 230000005684 electric field Effects 0.000 claims description 9
- 238000005452 bending Methods 0.000 claims description 6
- 230000000694 effects Effects 0.000 claims description 4
- 102000029749 Microtubule Human genes 0.000 claims description 3
- 108091022875 Microtubule Proteins 0.000 claims description 3
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 3
- 229910052782 aluminium Inorganic materials 0.000 claims description 3
- 230000008859 change Effects 0.000 claims description 3
- 239000004020 conductor Substances 0.000 claims description 3
- 102000013035 dynein heavy chain Human genes 0.000 claims description 3
- 108060002430 dynein heavy chain Proteins 0.000 claims description 3
- 210000004688 microtubule Anatomy 0.000 claims description 3
- 230000004899 motility Effects 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 230000009182 swimming Effects 0.000 claims description 3
- 238000004088 simulation Methods 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 239000013060 biological fluid Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000002032 lab-on-a-chip Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004377 microelectronic Methods 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/16—Microfluidic devices; Capillary tubes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/02—Photobioreactors
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
Abstract
The fluid driving method based on the algae cells specifically comprises the following steps: the viscous micropump comprises a fluid driving chip, an L-shaped adapter plate, a three-dimensional nano moving platform, a microscope, a CCD (charge coupled device), a high-precision Z-direction displacement platform, a projector, a focusing objective lens, a signal generator and a computer; (II) slowly injecting the treated algae cell solution into the micro-pipeline in the fluid driving chip from the fluid inlet; thirdly, projecting a light spot onto the lower surface of the fluid driving chip; fourthly, adjusting a focal plane of the focusing objective lens and a focal plane of the microscope; fifthly, adjusting the size of the light spot to enable the signal generator to generate a sine signal to act on the fluid driving chip; capturing autorotation algae cells in the micro-pipeline by using the light spots and moving the algae cells; and (seventh), removing the redundant algae cells from the micro-pipeline. The invention utilizes the rotating algae cells to complete the driving of the fluid, and does not need external extra energy supply.
Description
Technical Field
The invention relates to the technical field of biological fluid driving, in particular to a fluid driving method based on algae cells.
Background
Micro total analysis systems, also known as lab-on-a-chip, have since been proposed for widespread use in chemistry, life sciences and microelectronics. The micro total analysis system can be used as a micro filter for filtering and collecting micro particles, can also be used as a micro mixer for mixing samples and analytes in the fields of chemistry, biology and the like, and even can be used as a micro reactor for polymerase chain reaction. In order to realize the above-mentioned applications, in a micro total analysis system, it is often necessary to integrate some functional components, including: the flow sensor, the micro valve, the micropump, among these several functional components, the micropump is the most critical element, and the micropump can be used for driving the controllable transmission of fluid in the micro total analysis system.
Conventional micro-electromechanical system (MEMS) based micropumps can be divided into two broad categories, mechanical micropumps and non-mechanical micropumps. The viscous micropump belongs to an improved mechanical micropump, realizes the driving of fluid by utilizing a rotating cylinder at an asymmetric position of a microchannel, has the high efficiency of the traditional mechanical micropump, and also has the stability of a non-mechanical micropump. In viscous micropumps, however, it is a very challenging task to fabricate micro-actuators capable of 360 degree rotation.
Disclosure of Invention
The invention aims to provide a fluid driving method based on algae cells, which utilizes rotating algae cells to drive fluid without external additional energy supply.
In order to achieve the purpose, the invention adopts the following technical scheme:
the fluid driving method based on the algae cells specifically comprises the following steps: the viscous micropump comprises a fluid driving chip, an L-shaped adapter plate, a three-dimensional nanometer moving platform, a microscope, a CCD (charge coupled device), a high-precision Z-direction displacement platform, a projector, a focusing objective lens, a signal generator and a computer, wherein the fluid driving chip is horizontally arranged, a fluid inlet and a fluid outlet are arranged on the upper surface of the fluid driving chip at intervals from left to right, a micro pipeline positioned between the fluid inlet and the fluid outlet is arranged in the fluid driving chip along the left-right direction, the fluid inlet is connected with the left end of the micro pipeline, the fluid outlet is connected with the right end of the micro pipeline, the L-shaped adapter plate comprises a horizontal plate and a vertical plate, the left side of the horizontal plate of the L-shaped adapter plate is fixedly connected with the lower side of the vertical plate in an integrated forming manner, the left side of the vertical plate of the L-shaped adapter plate is fixedly connected to the three-dimensional nanometer moving platform, and the middle part of the right side of the horizontal plate of the L-shaped adapter plate is provided with an upper side, The fluid driving chip is matched and embedded in the upper groove of the stepped rectangular groove, the microscope is arranged right above the fluid driving chip, the CCD is fixedly arranged on the upper part of the microscope, a lens barrel of the microscope is fixedly connected with the high-precision Z-direction displacement table, the projector is arranged right below the fluid driving chip, a lens of the projector is arranged at the top of the projector and irradiates upwards, the focusing objective lens is arranged on the upper part of the lens of the projector, the light path of the focusing objective lens is superposed with the light path of the microscope, the signal generator is electrically connected with the fluid driving chip, and the computer is respectively in signal connection with the CCD and the projector;
(II) slowly injecting the treated algae cell solution into the micro-pipeline in the fluid driving chip from the fluid inlet;
drawing a plurality of light spot images on a computer by using flash drawing software, wherein each light spot image is projected upwards through a projector, converged through a focusing objective lens and then projected upwards through a gap of a stepped rectangular groove to the lower surface of the fluid driving chip;
adjusting the three-dimensional nano moving platform and the high-precision Z-direction displacement platform to enable the focal plane of the focusing objective lens and the focal plane of the microscope to be focused at the micro-pipeline inside the fluid driving chip;
fifthly, adjusting the size of the light spot drawn in the step (three), adjusting the size of the light spot finally projected on the fluid driving chip to 10-15 μm, and then starting a signal generator, wherein the signal generator generates a sinusoidal signal with the frequency of 60KHz and the peak value of 6V and acts on the fluid driving chip through a lead;
capturing autorotation algae cells in the micro-pipeline by utilizing the projected light spots and moving the algae cells for arrangement;
and (seventh), removing the redundant algae cells from the micro-pipe, and turning off the signal generator and the projector, so that the algae cells rotating in the micro-pipe drive the fluid to flow in the micro-pipe from the fluid inlet to the fluid outlet.
The fluid driving chip comprises an upper ITO glass layer and a lower ITO glass layer, a microstructure layer is arranged between the upper ITO glass layer and the lower ITO glass layer, the length directions of the upper ITO glass layer and the lower ITO glass layer are both arranged along the left-right direction, the upper ITO glass layer and the lower ITO glass layer are both in a rectangular sheet structure with the length of 25mm and the width of 20mm, the thickness of the upper ITO glass layer and the thickness of the lower ITO glass layer are both 1mm, two first through holes with the diameter of 0.5mm and through up and down are arranged on the upper ITO glass layer at intervals left and right, the first through hole on the left side is a fluid inlet, the first through hole on the right side is a fluid outlet, the microstructure layer is in a square sheet structure with the side length of 20mm, the thickness of the microstructure layer is 0.02-0.1mm, a rectangular long hole with the length of 10mm and the width of 0.1mm and through up and down is arranged in the middle of the microstructure layer along the left-right direction, two second through holes with the diameter of 0.5mm and through up and down are arranged on the microstructure layer at intervals left and right, the rectangular long hole is positioned between the two second through holes, the left end of the rectangular long hole is communicated with the second through hole on the left side, the right end of the rectangular long hole is communicated with the second through hole on the right side, the lower surface of the upper ITO glass layer and the upper surface of the lower ITO glass layer are conductive surfaces, the signal generator is respectively electrically connected with the two conductive surfaces through a wire, a hydrogenated amorphous silicon layer is spirally coated on the conductive surface of the lower ITO glass layer, the area of the hydrogenated amorphous silicon layer is a square area with the side length of 20mm, the thickness of the hydrogenated amorphous silicon layer is 1 mu m, the right side edge of the hydrogenated amorphous silicon layer is flush with the right side edge of the upper surface of the lower ITO glass layer, the conductive surface of the upper ITO glass layer is bonded with the upper surface of the microstructure layer, the left side edge of the lower surface of the upper ITO glass layer is flush with the left side edge of the upper surface of the microstructure layer, the upper surface of the hydrogenated amorphous layer is bonded with the lower surface of the microstructure layer, and the microstructure layer is aligned with the hydrogenated amorphous layer up and down, the two first through holes are respectively aligned with the two second through holes up and down correspondingly, and the rectangular long hole is surrounded into a micro pipeline with the length of 10mm, the width of 0.1mm and the height of 0.02-0.1mm after the upper ITO glass layer, the micro-structural layer and the lower ITO glass layer are bonded.
The L type keysets is aluminum plate, and the width of L type keysets is 50mm, and the thickness of L type keysets is 2mm, and the height of the vertical board of L type keysets is 50mm, and the length of the horizontal plate of L type keysets is 200mm, and the upper portion groove size of ladder rectangular channel is long 25mm, wide 20mm, thick 0.5mm, and the lower part groove size of ladder rectangular channel is long 24mm, wide 18mm, thick 1.5 mm.
The step (II) is specifically as follows: centrifuging 1ml of algae cell solution at 3000rpm for 6min, removing supernatant, adding deionized water, and adjusting the concentration of the centrifuged algae cell solution to 1 × 105cells/ml, and then injecting the treated algal cell solution from the fluid inlet into the microchannel within the fluid driven chip.
The reason why the size of the light spot finally projected onto the fluid driving chip is adjusted to 10-15 μm in the step (five) is as follows: the algae cells are elliptical or oval in shape and about 10 μm in size, and the light spot size is adjusted to be equivalent to the size of the algae cells, so that the capture operation of single algae cells is more convenient to realize;
and (V) the sinusoidal signal generated by the signal generator acts on the conductive surface of the upper ITO glass layer and the conductive surface of the lower ITO glass layer through the conducting wires respectively.
The step (six) is specifically as follows: feeding back images in the fluid driving chip observed by a microscope to a computer through a CCD, moving each light spot drawn by flash drawing software on the computer by combining the fed-back images, further moving the light spots projected onto the fluid driving chip, respectively capturing autorotation algae cells in the micro-pipeline by each light spot, arranging each clockwise rotation algae cell at a position close to the wall of the front side pipeline in the micro-pipeline at a left-right interval, and arranging each anticlockwise rotation algae cell at a position close to the wall of the rear side pipeline in the micro-pipeline at a left-right interval.
The rotating mechanism of the algae cells in the step (six) is as follows: the front end of the body of the algae cell is provided with two flagella, the two flagella of the algae cell belong to a linear motor and can only generate a certain degree of bending swing, the bending swing of the two flagella of the algae cell is generated by relative sliding caused by the conformational change of dynein between adjacent microtubules, when both the two flagella are in a free state, the algae cell shows a random swimming motion form, and when only a single flagella is in the free swing state, the algae cell shows a counterclockwise rotation or clockwise rotation motion form.
The mechanism of capturing and moving algae cells by the light spots in the step (six) is as follows: when no light spot irradiates the ITO glass layer, the conductivity of the hydrogenated amorphous silicon layer is very low, the hydrogenated amorphous silicon layer can be regarded as an insulating layer, the voltage loaded on the fluid driving chip is basically applied to the hydrogenated amorphous silicon layer, when the light spot irradiates the hydrogenated amorphous silicon layer, the conductivity of the hydrogenated amorphous silicon layer in a spot area is increased rapidly due to electron hole pairs generated by the light spot, the hydrogenated amorphous silicon layer in the spot area is equivalent to a conductor, most of the voltage falls on the microstructure layer, a non-uniform electric field is generated in the microstructure layer in the spot area and the spot-free area, algae cells are polarized in the non-uniform electric field area and further subjected to light-induced dielectrophoresis force, and the expression of the light-induced dielectrophoresis force borne by the algae cells is as follows:
in the formula
Is the dielectric constant of the solution medium,
is the diameter of the algal cells and is,
is a C-M factor
The real part of (a) is,
a gradient that is the square of the root mean square value of the electric field;
when the signal generator generates a sinusoidal signal with the frequency of 60KHz and the peak-to-peak value of 6V and acts on the conductive surface of the upper ITO glass layer and the conductive surface of the lower ITO glass layer of the fluid driving chip through the conducting wire, the algae cells are subjected to light-induced dielectrophoresis force of the facula-free area pointing to the facula area, and the light-induced dielectrophoresis force is larger than the own motility force of the algae cells, so that the facula can capture and move the algae cells.
The step (VII) is specifically as follows: locking the arranged algae cells by using each light spot, and injecting deionized water into the micro-pipeline from the fluid inlet at a certain flow rate, wherein the flow rate of the deionized water meets the following conditions: the fluid drag force on the algae cells locked by the light spots is required to be smaller than the light-induced dielectrophoresis force on the algae cells, so that the rest algae cells which are not locked by the light spots are washed away from the micro-pipe by the deionized water, and then the signal generator and the projector are closed, so that the algae cells rotating in the micro-pipe drive the fluid to flow from the fluid inlet to the fluid outlet.
The mechanism that the algae cells rotating in the micro-pipe drive the fluid to flow from the fluid inlet to the fluid outlet in the micro-pipe in the step (seven) is as follows: because the clearance of the algae cell of rotation and the pipeline wall of rear side in the microchannel is different with its clearance with the pipeline wall of front side in the microchannel, then can cause the algae cell to rotate the in-process, the fluid viscous resistance that the algae cell both sides received is different, will produce a net flow to the fluid when the algae cell is rotatory like this, drives the fluid and can follow the left end of microchannel and flow right end: the row of the algae cells rotating anticlockwise close to the wall of the rear side pipeline in the micro-pipeline can drive the fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, the row of the algae cells rotating clockwise close to the wall of the front side pipeline in the micro-pipeline can also drive the fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, and the driving effects generated by the algae cells rotating in opposite directions on the two sides are consistent.
Compared with the prior art, the invention has outstanding substantive characteristics and remarkable progress, and particularly, the invention takes the rotating algae cells as functional components in the traditional micropump, thereby not only avoiding the difficulty of manufacturing a precise micro actuating mechanism, but also solving the problem of energy supply, completing the driving of fluid by utilizing the self energy of biological cells, not needing external extra energy supply, utilizing the cooperative work of a plurality of algae cells, and further improving the performance of the whole viscous micropump.
Drawings
Fig. 1 is a schematic view of the construction of a viscous micro-pump according to the present invention.
Fig. 2 is a schematic structural diagram of an L-shaped adapter plate of the present invention.
Fig. 3 is a schematic structural diagram of a fluid driving chip according to the present invention.
Fig. 4 is a schematic structural view of a microstructure layer of the present invention.
FIG. 5 is a schematic structural view of a lower ITO glass layer of the present invention.
FIG. 6 is a schematic diagram showing the distribution of algal cells according to the present invention in the microchannel of the fluid driver chip.
FIG. 7 is a view of a fluid driving chip of the present invention
And (5) finite element simulation results.
FIG. 8 is a finite element simulation result of the algae cell driving fluid rotating in opposite directions inside the microchannel according to the present invention.
Detailed Description
The embodiments of the present invention are further described below with reference to the drawings.
As shown in fig. 1-6, a fluid driving method based on algae cells, (i) a viscous micropump is designed, the viscous micropump includes a fluid driving chip 1, an L-shaped adapter plate 2, a three-dimensional nano moving platform 3, a microscope 4, a CCD5, a high-precision Z-displacement stage 6, a projector 7, a focusing objective lens 8, a signal generator 9 and a computer 10, the fluid driving chip 1 is horizontally arranged, a fluid inlet and a fluid outlet are arranged on the upper surface of the fluid driving chip 1 at intervals from left to right, a micro-pipe 11 located between the fluid inlet and the fluid outlet is arranged in the fluid driving chip 1 along the left-right direction, the fluid inlet is connected with the left end of the micro-pipe 11, the fluid outlet is connected with the right end of the micro-pipe 11, the L-shaped adapter plate 2 includes a horizontal plate and a vertical plate, the left side of the horizontal plate of the L-shaped adapter plate 2 is fixedly connected with the lower side of the vertical plate by integral molding, the left side surface of a vertical plate of an L-shaped adapter plate 2 is fixedly connected on a three-dimensional nano moving platform 3, the middle part of the right side of a horizontal plate of the L-shaped adapter plate 2 is provided with a stepped rectangular groove 12 with an upper side, a lower side and a right side which are open and a big upper part and a small lower part, the fluid driving chip 1 and the upper part of the stepped rectangular groove 12 have the same size, the fluid driving chip 1 is embedded in the upper part of the stepped rectangular groove 12 in a matching way, a microscope 4 is arranged right above the fluid driving chip 1, a CCD5 is fixedly arranged on the upper part of the microscope 4, a lens barrel of the microscope 4 is fixedly connected with a high-precision Z-direction displacement table 6, a projector 7 is arranged right below the fluid driving chip 1, a lens of the projector 7 is arranged at the top of the projector 7 and irradiates upwards, a focusing objective 8 is arranged on the upper part of the lens of the projector 7, the light path of the focusing objective 8 is superposed with the light path of the microscope 4, a signal generator 9 is electrically connected with the fluid driving chip 1, the computer 10 is respectively connected with the CCD5 and the projector 7 through signals;
(II) slowly injecting the treated algae cell solution into the micro-pipeline 11 in the fluid driving chip 1 from the fluid inlet;
thirdly, drawing a plurality of light spot images on a computer 10 by using flash drawing software, wherein each light spot image is projected upwards through a projector 7, converged through a focusing objective 8 and projected upwards through a gap of a stepped rectangular groove 12 to the lower surface of the fluid driving chip 1;
fourthly, adjusting the three-dimensional nano moving platform 3 and the high-precision Z-direction displacement platform 6 to enable the focal plane of the focusing objective lens 8 and the focal plane of the microscope 4 to be focused on the micro-pipeline 11 in the fluid driving chip 1;
fifthly, adjusting the size of the light spot drawn in the step (three), adjusting the size of the light spot finally projected on the fluid driving chip 1 to be 10-15 mu m, and then starting the signal generator 9, wherein the signal generator 9 generates a sinusoidal signal with the frequency of 60KHz and the peak value of 6V and acts on the fluid driving chip 1 through a lead;
capturing autorotation algae cells in the micro-pipeline 11 by utilizing the projected light spots and moving the algae cells for arrangement;
and (seventh), removing the redundant algae cells from the microchannel 11, and turning off the signal generator 9 and the projector 7, so that the algae cells rotating in the microchannel 11 drive the fluid to flow in the microchannel 11 from the fluid inlet to the fluid outlet.
The fluid driving chip 1 comprises an upper ITO glass layer 13 and a lower ITO glass layer 14, a micro-structural layer 15 is arranged between the upper ITO glass layer 13 and the lower ITO glass layer 14, the length directions of the upper ITO glass layer 13 and the lower ITO glass layer 14 are both arranged along the left and right direction, the upper ITO glass layer 13 and the lower ITO glass layer 14 are both of rectangular sheet structures with the length of 25mm and the width of 20mm, the thickness of the upper ITO glass layer 13 and the thickness of the lower ITO glass layer 14 are both 1mm, two first through holes 16 which are 0.5mm in diameter and are through up and down are arranged on the upper ITO glass layer 13 at intervals from left to right, the first through hole 16 on the left side is a fluid inlet, the first through hole 16 on the right side is a fluid outlet, the micro-structural layer 15 is of a square sheet structure with the side length of 20mm, the thickness of the micro-structural layer 15 is 0.02-0.1mm, a rectangular long hole 17 which is 10mm in length, 0.1mm in width and is through up and down is arranged in the middle of the left and right direction of the micro-structural layer 15, two second through holes 18 which are through up and down are arranged at intervals from left to right, the rectangular long hole 17 is positioned between the two second through holes 18, the left end of the rectangular long hole 17 is communicated with the left second through hole 18, the right end of the rectangular long hole 17 is communicated with the right second through hole 18, the lower surface of the upper ITO glass layer 13 and the upper surface of the lower ITO glass layer 14 are conductive surfaces, the signal generator 9 is electrically connected with the two conductive surfaces through wires respectively, the conductive surface of the lower ITO glass layer 14 is coated with a hydrogenated amorphous silicon layer 19 in a spinning mode, the area of the hydrogenated amorphous silicon layer 19 is a square area with the side length of 20mm, the thickness of the hydrogenated amorphous silicon layer 19 is 1 mu m, the right side edge of the hydrogenated amorphous silicon layer 19 is flush with the right side edge of the upper surface of the lower ITO glass layer 14, the conductive surface of the upper ITO glass layer 13 is bonded with the upper surface of the microstructure layer 15, the left side edge of the lower surface of the upper ITO glass layer 13 is flush with the left side edge of the upper surface of the microstructure layer 15, the upper surface of the hydrogenated amorphous silicon layer 19 is bonded with the lower surface of the microstructure layer 15, the microstructure layer 15 and the hydrogenated amorphous silicon layer 19 are aligned up and down, the two first through holes 16 are respectively aligned up and down with the two second through holes 18 correspondingly, and the rectangular long hole 17 is surrounded into the micro-pipeline 11 with the length of 10mm, the width of 0.1mm and the height of 0.02-0.1mm after the upper ITO glass layer 13, the microstructure layer 15 and the lower ITO glass layer 14 are bonded.
The step (II) is specifically as follows: centrifuging 1ml of algae cell solution at 3000rpm for 6min, removing supernatant, adding deionized water, and adjusting the concentration of the centrifuged algae cell solution to 1 × 105cells/ml, and then the treated algal cell solution is injected from the fluid inlet into the microchannel 11 within the fluid driven chip 1.
The reason why the spot size finally projected onto the fluid driving chip 1 is adjusted to 10 to 15 μm in the step (five) is as follows: the algae cells are elliptical or oval in shape and about 10 μm in size, and the light spot size is adjusted to be equivalent to the size of the algae cells, so that the capture operation of single algae cells is more convenient to realize;
in the step (five), the sinusoidal signal generated by the signal generator 9 acts on the conductive surface of the upper ITO glass layer 13 and the conductive surface of the lower ITO glass layer 14 through the conducting wires respectively.
The step (six) is specifically as follows: feeding back the image in the fluid driving chip 1 observed by the microscope 4 to the computer 10 through the CCD5, moving each light spot drawn by flash drawing software on the computer 10 by combining the fed-back image, further moving the light spot projected on the fluid driving chip 1, capturing the autorotation algae cells in the micro-pipeline 11 by each light spot, arranging each clockwise rotation algae cell at the position close to the front side pipeline wall in the micro-pipeline 11 at left and right intervals, and arranging each counterclockwise rotation algae cell at the position close to the rear side pipeline wall in the micro-pipeline 11 at left and right intervals.
The rotating mechanism of the algae cells in the step (six) is as follows: the front end of the body of the algae cell is provided with two flagella, the two flagella of the algae cell belong to a linear motor and can only generate a certain degree of bending swing, the bending swing of the two flagella of the algae cell is generated by relative sliding caused by the conformational change of dynein between adjacent microtubules, when both the two flagella are in a free state, the algae cell shows a random swimming motion form, and when only a single flagella is in the free swing state, the algae cell shows a counterclockwise rotation or clockwise rotation motion form.
The mechanism of capturing and moving algae cells by the light spots in the step (six) is as follows: when the ITO glass layer 14 is not irradiated by the light spot, the conductivity of the hydrogenated amorphous silicon layer 19 is very low, and the hydrogenated amorphous silicon layer 19 can be regarded as an insulating layer, the voltage loaded on the fluid driving chip 1 is basically applied to the hydrogenated amorphous silicon layer 19, when the light spot is irradiated on the hydrogenated amorphous silicon layer 19, the conductivity of the hydrogenated amorphous silicon layer 19 in the spot area is increased sharply due to electron-hole pairs generated by the light spot, the hydrogenated amorphous silicon layer 19 in the spot area is equivalent to a conductor, most of the voltage falls on the microstructure layer 15, so that a non-uniform electric field is generated in the microstructure layer 15 in the spot area and the spot-free area, algae cells are polarized in the non-uniform electric field area and are further subjected to light-induced electrophoresis dielectric force, and the expression of the light-induced dielectric force on the algae cells is as follows:
in the formula
Is the dielectric constant of the solution medium,
is the diameter of the algal cells and is,
is a C-M factor
The real part of (a) is,
a gradient that is the square of the root mean square value of the electric field;
when the signal generator 9 generates a sinusoidal signal with a frequency of 60KHz and a peak-to-peak value of 6V and acts on the conductive surface of the upper ITO glass layer 13 and the conductive surface of the lower ITO glass layer 14 of the fluid driving chip 1 through the wires, the algae cells are subjected to a light-induced dielectrophoresis force of the spot-free region directed to the spot region, and the light-induced dielectrophoresis force is greater than the motility force of the algae cells themselves, so that the spots can capture and move the algae cells.
The step (VII) is specifically as follows: locking the arranged algae cells by using each light spot, and injecting deionized water into the micro-pipeline 11 from the fluid inlet at a certain flow rate, wherein the flow rate of the deionized water meets the following conditions: the fluid drag force on the algae cells locked by the light spot is smaller than the light-induced dielectrophoresis force on the algae cells, so that the deionized water flushes the rest of the algae cells which are not locked by the light spot away from the microchannel 11, and then the signal generator 9 and the projector 7 are closed, so that the algae cells rotating in the microchannel 11 drive the fluid to flow from the fluid inlet to the fluid outlet.
The mechanism that the algae cells rotating in the micro-pipe 11 in the step (seven) drive the fluid to flow in the micro-pipe 11 from the fluid inlet to the fluid outlet is as follows: because the gap between the autorotation algae cell and the inner rear side pipeline wall of the micro pipeline 11 is different from the gap between the autorotation algae cell and the inner front side pipeline wall of the micro pipeline 11, the viscous resistance of the fluid on two sides of the algae cell is different in the rotating process of the algae cell, so that the algae cell can generate a net flow to the fluid when rotating, and the fluid can flow to the right end from the left end of the micro pipeline 11: the counter-clockwise rotating row of algae cells near the rear wall of the microchannel 11 drives the fluid to flow from the fluid inlet to the fluid outlet in the microchannel 11, the counter-clockwise rotating row of algae cells near the front wall of the microchannel 11 also drives the fluid to flow from the fluid inlet to the fluid outlet in the microchannel 11, and the driving effects of the algae cells rotating in opposite directions on the two sides are the same.
FIG. 7 shows that when a sinusoidal signal of 60KHz and 6V peak value is applied to the conductive surface of the upper ITO glass layer 13 and the conductive surface of the lower ITO glass layer 14 of the fluid driving chip 1, and the spot size projected onto the surface of the hydrogenated amorphous silicon layer 19 is 10 μm, the fluid driving chip 1 has a light spot size
Finite element simulation results, the magnitude and direction of which represent the magnitude and direction of light-induced dielectrophoretic force, black arrows in the simulation results
And the direction of most arrows is pointed to the facula area from the facula-free area from the simulation result.
FIG. 8 shows the result of finite element simulation of the fluid driving chip 1 driving the fluid by the algae cells rotating counterclockwise at 6.28rad/s near the inner rear side of the micro duct 11 and the algae cells rotating clockwise at 6.28rad/s near the inner front side of the micro duct 11, and it can be seen that the driving effect generated by the algae cells rotating in opposite directions at both sides is the same.
The above embodiments are merely to illustrate rather than to limit the technical solutions of the present invention, and although the present invention has been described in detail with reference to the above embodiments, those of ordinary skill in the art should understand that; modifications and equivalents may be made thereto without departing from the spirit and scope of the invention and it is intended to cover in the claims the invention as defined in the appended claims.
Claims (10)
1. A method of fluid driving based on algal cells, characterized by: the method specifically comprises the following steps:
the viscous micropump comprises a fluid driving chip, an L-shaped adapter plate, a three-dimensional nanometer moving platform, a microscope, a CCD (charge coupled device), a high-precision Z-direction displacement platform, a projector, a focusing objective lens, a signal generator and a computer, wherein the fluid driving chip is horizontally arranged, a fluid inlet and a fluid outlet are arranged on the upper surface of the fluid driving chip at intervals from left to right, a micro pipeline positioned between the fluid inlet and the fluid outlet is arranged in the fluid driving chip along the left-right direction, the fluid inlet is connected with the left end of the micro pipeline, the fluid outlet is connected with the right end of the micro pipeline, the L-shaped adapter plate comprises a horizontal plate and a vertical plate, the left side of the horizontal plate of the L-shaped adapter plate is fixedly connected with the lower side of the vertical plate in an integrated forming manner, the left side of the vertical plate of the L-shaped adapter plate is fixedly connected to the three-dimensional nanometer moving platform, and the middle part of the right side of the horizontal plate of the L-shaped adapter plate is provided with an upper side, The fluid driving chip is matched and embedded in the upper groove of the stepped rectangular groove, the microscope is arranged right above the fluid driving chip, the CCD is fixedly arranged on the upper part of the microscope, a lens barrel of the microscope is fixedly connected with the high-precision Z-direction displacement table, the projector is arranged right below the fluid driving chip, a lens of the projector is arranged at the top of the projector and irradiates upwards, the focusing objective lens is arranged on the upper part of the lens of the projector, the light path of the focusing objective lens is superposed with the light path of the microscope, the signal generator is electrically connected with the fluid driving chip, and the computer is respectively in signal connection with the CCD and the projector;
(II) slowly injecting the treated algae cell solution into the micro-pipeline in the fluid driving chip from the fluid inlet;
drawing a plurality of light spot images on a computer by using flash drawing software, wherein each light spot image is projected upwards through a projector, converged through a focusing objective lens and then projected upwards through a gap of a stepped rectangular groove to the lower surface of the fluid driving chip;
adjusting the three-dimensional nano moving platform and the high-precision Z-direction displacement platform to enable the focal plane of the focusing objective lens and the focal plane of the microscope to be focused at the micro-pipeline inside the fluid driving chip;
fifthly, adjusting the size of the light spot drawn in the step (three), adjusting the size of the light spot finally projected on the fluid driving chip to 10-15 μm, and then starting a signal generator, wherein the signal generator generates a sinusoidal signal with the frequency of 60KHz and the peak value of 6V and acts on the fluid driving chip through a lead;
capturing autorotation algae cells in the micro-pipeline by utilizing the projected light spots and moving the algae cells for arrangement;
and (seventh), removing the redundant algae cells from the micro-pipe, and turning off the signal generator and the projector, so that the algae cells rotating in the micro-pipe drive the fluid to flow in the micro-pipe from the fluid inlet to the fluid outlet.
2. The algae cell-based fluid driven method of claim 1, wherein: the fluid driving chip comprises an upper ITO glass layer and a lower ITO glass layer, a microstructure layer is arranged between the upper ITO glass layer and the lower ITO glass layer, the length directions of the upper ITO glass layer and the lower ITO glass layer are both arranged along the left-right direction, the upper ITO glass layer and the lower ITO glass layer are both in a rectangular sheet structure with the length of 25mm and the width of 20mm, the thickness of the upper ITO glass layer and the thickness of the lower ITO glass layer are both 1mm, two first through holes with the diameter of 0.5mm and through up and down are arranged on the upper ITO glass layer at intervals left and right, the first through hole on the left side is a fluid inlet, the first through hole on the right side is a fluid outlet, the microstructure layer is in a square sheet structure with the side length of 20mm, the thickness of the microstructure layer is 0.02-0.1mm, a rectangular long hole with the length of 10mm and the width of 0.1mm and through up and down is arranged in the middle of the microstructure layer along the left-right direction, two second through holes with the diameter of 0.5mm and through up and down are arranged on the microstructure layer at intervals left and right, the rectangular long hole is positioned between the two second through holes, the left end of the rectangular long hole is communicated with the second through hole on the left side, the right end of the rectangular long hole is communicated with the second through hole on the right side, the lower surface of the upper ITO glass layer and the upper surface of the lower ITO glass layer are conductive surfaces, the signal generator is respectively electrically connected with the two conductive surfaces through a wire, a hydrogenated amorphous silicon layer is spirally coated on the conductive surface of the lower ITO glass layer, the area of the hydrogenated amorphous silicon layer is a square area with the side length of 20mm, the thickness of the hydrogenated amorphous silicon layer is 1 mu m, the right side edge of the hydrogenated amorphous silicon layer is flush with the right side edge of the upper surface of the lower ITO glass layer, the conductive surface of the upper ITO glass layer is bonded with the upper surface of the microstructure layer, the left side edge of the lower surface of the upper ITO glass layer is flush with the left side edge of the upper surface of the microstructure layer, the upper surface of the hydrogenated amorphous layer is bonded with the lower surface of the microstructure layer, and the microstructure layer is aligned with the hydrogenated amorphous layer up and down, the two first through holes are respectively aligned with the two second through holes up and down correspondingly, and the rectangular long hole is surrounded into a micro pipeline with the length of 10mm, the width of 0.1mm and the height of 0.02-0.1mm after the upper ITO glass layer, the micro-structural layer and the lower ITO glass layer are bonded.
3. The algae cell-based fluid driven method of claim 2, wherein: the L type keysets is aluminum plate, and the width of L type keysets is 50mm, and the thickness of L type keysets is 2mm, and the height of the vertical board of L type keysets is 50mm, and the length of the horizontal plate of L type keysets is 200mm, and the upper portion groove size of ladder rectangular channel is long 25mm, wide 20mm, thick 0.5mm, and the lower part groove size of ladder rectangular channel is long 24mm, wide 18mm, thick 1.5 mm.
4. The algae cell-based fluid driven method of claim 3, wherein: the step (II) is specifically as follows: centrifuging 1ml of algae cell solution at 3000rpm for 6min, removing supernatant, adding deionized water, and adjusting the concentration of the centrifuged algae cell solution to 1 × 105cells/ml, and then injecting the treated algal cell solution from the fluid inlet into the microchannel within the fluid driven chip.
5. The algae cell-based fluid driven method of claim 4, wherein: the reason why the size of the light spot finally projected onto the fluid driving chip is adjusted to 10-15 μm in the step (five) is as follows: the algae cells are elliptical or oval in shape and about 10 μm in size, and the light spot size is adjusted to be equivalent to the size of the algae cells, so that the capture operation of single algae cells is more convenient to realize;
and (V) the sinusoidal signal generated by the signal generator acts on the conductive surface of the upper ITO glass layer and the conductive surface of the lower ITO glass layer through the conducting wires respectively.
6. The algae cell-based fluid driven method of claim 5, wherein: the step (six) is specifically as follows: feeding back images in the fluid driving chip observed by a microscope to a computer through a CCD, moving each light spot drawn by flash drawing software on the computer by combining the fed-back images, further moving the light spots projected onto the fluid driving chip, respectively capturing autorotation algae cells in the micro-pipeline by each light spot, arranging each clockwise rotation algae cell at a position close to the wall of the front side pipeline in the micro-pipeline at a left-right interval, and arranging each anticlockwise rotation algae cell at a position close to the wall of the rear side pipeline in the micro-pipeline at a left-right interval.
7. The algae cell-based fluid driven method of claim 6, wherein: the rotating mechanism of the algae cells in the step (six) is as follows: the front end of the body of the algae cell is provided with two flagella, the two flagella of the algae cell belong to a linear motor and can only generate a certain degree of bending swing, the bending swing of the two flagella of the algae cell is generated by relative sliding caused by the conformational change of dynein between adjacent microtubules, when both the two flagella are in a free state, the algae cell shows a random swimming motion form, and when only a single flagella is in the free swing state, the algae cell shows a counterclockwise rotation or clockwise rotation motion form.
8. The algae cell-based fluid driven method of claim 7, wherein: the mechanism of capturing and moving algae cells by the light spots in the step (six) is as follows: when no light spot irradiates the ITO glass layer, the conductivity of the hydrogenated amorphous silicon layer is very low, the hydrogenated amorphous silicon layer can be regarded as an insulating layer, the voltage loaded on the fluid driving chip is basically applied to the hydrogenated amorphous silicon layer, when the light spot irradiates the hydrogenated amorphous silicon layer, the conductivity of the hydrogenated amorphous silicon layer in a spot area is increased rapidly due to electron hole pairs generated by the light spot, the hydrogenated amorphous silicon layer in the spot area is equivalent to a conductor, most of the voltage falls on the microstructure layer, a non-uniform electric field is generated in the microstructure layer in the spot area and the spot-free area, algae cells are polarized in the non-uniform electric field area and further subjected to light-induced dielectrophoresis force, and the expression of the light-induced dielectrophoresis force borne by the algae cells is as follows:
in the formula
Is the dielectric constant of the solution medium,
is the diameter of the algal cells and is,
is a C-M factor
The real part of (a) is,
a gradient that is the square of the root mean square value of the electric field;
when the signal generator generates a sinusoidal signal with the frequency of 60KHz and the peak-to-peak value of 6V and acts on the conductive surface of the upper ITO glass layer and the conductive surface of the lower ITO glass layer of the fluid driving chip through the conducting wire, the algae cells are subjected to light-induced dielectrophoresis force of the facula-free area pointing to the facula area, and the light-induced dielectrophoresis force is larger than the own motility force of the algae cells, so that the facula can capture and move the algae cells.
9. The algae cell-based fluid driven method of claim 8, wherein: the step (VII) is specifically as follows: locking the arranged algae cells by using each light spot, and injecting deionized water into the micro-pipeline from the fluid inlet at a certain flow rate, wherein the flow rate of the deionized water meets the following conditions: the fluid drag force on the algae cells locked by the light spots is required to be smaller than the light-induced dielectrophoresis force on the algae cells, so that the rest algae cells which are not locked by the light spots are washed away from the micro-pipe by the deionized water, and then the signal generator and the projector are closed, so that the algae cells rotating in the micro-pipe drive the fluid to flow from the fluid inlet to the fluid outlet.
10. The algae cell-based fluid driven method of claim 9, wherein: the mechanism that the algae cells rotating in the micro-pipe drive the fluid to flow from the fluid inlet to the fluid outlet in the micro-pipe in the step (seven) is as follows: because the clearance of the algae cell of rotation and the pipeline wall of rear side in the microchannel is different with its clearance with the pipeline wall of front side in the microchannel, then can cause the algae cell to rotate the in-process, the fluid viscous resistance that the algae cell both sides received is different, will produce a net flow to the fluid when the algae cell is rotatory like this, drives the fluid and can follow the left end of microchannel and flow right end: the row of the algae cells rotating anticlockwise close to the wall of the rear side pipeline in the micro-pipeline can drive the fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, the row of the algae cells rotating clockwise close to the wall of the front side pipeline in the micro-pipeline can also drive the fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, and the driving effects generated by the algae cells rotating in opposite directions on the two sides are consistent.
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