CN114107025B - Fluid driving method based on algae cells - Google Patents
Fluid driving method based on algae cells Download PDFInfo
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- CN114107025B CN114107025B CN202111439893.6A CN202111439893A CN114107025B CN 114107025 B CN114107025 B CN 114107025B CN 202111439893 A CN202111439893 A CN 202111439893A CN 114107025 B CN114107025 B CN 114107025B
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- 239000012530 fluid Substances 0.000 title claims abstract description 159
- 241000195493 Cryptophyta Species 0.000 title claims abstract description 140
- 238000000034 method Methods 0.000 title claims abstract description 20
- 238000006073 displacement reaction Methods 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 146
- 239000011521 glass Substances 0.000 claims description 75
- 229910021417 amorphous silicon Inorganic materials 0.000 claims description 37
- 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
- 230000005684 electric field Effects 0.000 claims description 9
- 230000000149 penetrating effect Effects 0.000 claims description 9
- 238000005452 bending Methods 0.000 claims description 6
- 230000009182 swimming Effects 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
- 239000004020 conductor Substances 0.000 claims description 3
- 210000004688 microtubule Anatomy 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000004088 simulation Methods 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 4
- 239000013060 biological fluid Substances 0.000 description 1
- 238000010586 diagram 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
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000003752 polymerase chain reaction 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
A method of algae cell-based fluid driving, comprising the steps of: firstly, designing a viscous micropump, wherein the viscous micropump comprises a fluid driving chip, an L-shaped adapter plate, a three-dimensional nano moving platform, a microscope, a CCD, a high-precision Z-direction displacement platform, a projector, a focusing objective lens, a signal generator and a computer; slowly injecting the treated algae cell solution into the micro-pipeline in the fluid driving chip from the fluid inlet; (III) projecting a light spot onto the lower surface of the fluid drive chip; fourth, adjust the focal plane of the focusing objective lens and the 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 algae cells rotating in the micro-pipeline by utilizing the light spots and moving the algae cells; and (seventh), removing redundant algae cells from the micro-pipeline. The invention utilizes the rotating algae cells to complete the driving of the fluid without the need of external additional energy.
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 been widely used in chemistry, life sciences, and microelectronics since being proposed. The micro total analysis system can be used as a micro filter for filtering and collecting micro particles, can be used as a micro mixer for mixing samples and analytes in the fields of chemistry, biology and the like, and can even be used as a micro reactor for polymerase chain reaction. In order to achieve the above mentioned applications, in micro total analysis systems, it is often also necessary to integrate some functional components, including: flow sensors, micro-valves, micro-pumps, among the several functional components, are the most critical elements with which the controllable transport of fluids in a micro-total analysis system can be driven.
Conventional micro-electromechanical system (MEMS) based micro-pumps can be divided into two main categories, mechanical micro-pumps and non-mechanical micro-pumps. The viscous micropump belongs to an improved mechanical micropump, utilizes a rotary cylinder at an asymmetric position of a microchannel to drive fluid, has the high efficiency of the traditional mechanical micropump, and has the stability of a non-mechanical micropump. However, in viscous micropumps, it is a very challenging task to make micro-actuators capable of 360 degrees of rotation.
Disclosure of Invention
The invention aims to provide a fluid driving method based on algae cells, which utilizes the rotating algae cells to complete the driving of the fluid without providing energy externally.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a method of algae cell-based fluid driving, comprising the steps of: the method comprises the steps of (a) designing a viscous micropump, wherein the viscous micropump comprises a fluid driving chip, an L-shaped adapter plate, a three-dimensional nano movable 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;
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 by a focusing objective lens and then projected onto the lower surface of the fluid driving chip through a gap of a stepped rectangular groove upwards;
fourthly, adjusting the three-dimensional nano moving platform and the high-precision Z-direction moving platform to ensure that the focal plane of the focusing objective lens and the focal plane of the microscope are focused at the micro-channel inside the fluid driving chip;
fifthly, adjusting the size of the light spot drawn in the step (III), adjusting the size of the light spot finally projected onto the fluid driving chip to be 10-15 mu m, starting a signal generator, generating a sinusoidal signal with the frequency of 60KHz and the peak-to-peak value of 6V by the signal generator, and acting on the fluid driving chip through a lead;
capturing algae cells rotating in the micro-pipeline by utilizing the projected light spots and moving the algae cells to be arranged;
and (seventh), removing redundant algae cells from the micro-pipeline, and turning off the signal generator and the projector, so that the algae cells rotating in the micro-pipeline drive fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline.
The fluid driving chip comprises an upper ITO glass layer and a lower ITO glass layer, a micro-structure 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 all arranged along the left-right direction, the upper ITO glass layer and the lower ITO glass layer are rectangular sheet structures 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 penetrating up and down are arranged on the upper ITO glass layer at intervals left and right, the left first through hole is a fluid inlet, the right first through hole is a fluid outlet, the micro-structure layer is a square sheet structure with the side length of 20mm, the thickness of the micro-structure layer is 0.02-0.1mm, a rectangular long hole with the length of 10mm and the width of 0.1mm and penetrating up and down is arranged in the left-right direction in the middle of the micro-structure layer, two second through holes with the diameter of 0.5mm and penetrating up and down are arranged on the micro-structure 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 left second through hole, the right end of the rectangular long hole is communicated with the right second through hole, 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 wires, the conductive surface of the lower ITO glass layer is spin-coated with a hydrogenated amorphous silicon 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 flush 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 silicon layer is flush with the lower surface of the microstructure layer, the microstructure layer and the hydrogenated amorphous silicon layer are vertically aligned, the two first through holes are respectively aligned with the two second through holes vertically, and the upper ITO glass layer, the micro-structure layer and the lower ITO glass layer are bonded to enclose the rectangular long holes into micro-pipes with the length of 10mm, the width of 0.1mm and the height of 0.02-0.1 mm.
The L-shaped adapter plate is an aluminum plate, the width of the L-shaped adapter plate is 50mm, the thickness of the L-shaped adapter plate is 2mm, the height of a vertical plate of the L-shaped adapter plate is 50mm, the length of a horizontal plate of the L-shaped adapter plate is 200mm, the upper groove size of the stepped rectangular groove is 25mm long, 20mm wide and 0.5mm thick, and the lower groove size of the stepped rectangular groove is 24mm long, 18mm wide and 1.5mm thick.
The second step is specifically as follows: taking 1ml of algae cell solution, centrifuging at 3000rpm in a centrifuge for 6min, removing supernatant, adding deionized water again, and adjusting the concentration of the centrifuged algae cell solution to 1×10 5 cells/ml, then the treated algae cell solution is injected from the fluid inlet into the micro-tubing within the fluid-driven chip.
The reason why the spot size finally projected onto the fluid driving chip is adjusted to 10-15 μm in the step (five) is that: the shape of the algae cells is elliptical or oval, the size is about 10 mu m, and the light spot size is adjusted to be equivalent to the algae cell size, so that the capturing operation of single algae cells is more convenient to realize;
and (V) the sinusoidal signals generated by the signal generator in the step (V) are respectively acted on the conductive surface of the upper ITO glass layer and the conductive surface of the lower ITO glass layer through wires.
The step (six) is specifically as follows: the image in the fluid driving chip observed by the microscope is fed back to the computer through the CCD, each light spot drawn by flash drawing software on the computer is moved in combination with the fed back image, then the light spot projected onto the fluid driving chip is moved, each light spot respectively captures algae cells rotating in the micro-pipeline, each clockwise rotated algae cell is arranged at a position close to the pipeline wall at the front side in the micro-pipeline at a left-right interval, and each anticlockwise rotated algae cell is arranged at a position close to the pipeline wall at the rear side in the micro-pipeline at a left-right interval.
The algae cell rotation mechanism 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 bending swing to a certain extent, the bending swing of the two flagella of the algae cell is generated by virtue of relative sliding caused by the conformational change of the dynamic protein between adjacent microtubules, when the two flagella are in a free state, the algae cell shows a random swimming movement form, and when only one flagellum is in the free swing, the algae cell shows a counterclockwise rotation or clockwise rotation movement 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, at this time, 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 at the light spot area is sharply increased due to electron hole pairs generated by the light spot, the hydrogenated amorphous silicon layer at the light spot area is equivalent to a conductor, most of the voltage falls on the microstructure layer, thus a non-uniform electric field is generated in the microstructure layer with and without the light spot area, alga cells are polarized in the non-uniform electric field area and then receive photoinduction dielectrophoresis force, and the expression of the photoinduction force received by the alga cells is as follows:
in the middle ofIs the dielectric constant of the solution medium, +.>Is the diameter of algae cells, < > and->Is C-M factorReal part of->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 wires, the algae cells are subjected to light-induced dielectrophoresis force with the spot-free area pointing to the spot area, and the light-induced dielectrophoresis force is larger than the swimming force of the algae cells, so that the light spots can capture and move the algae cells.
The step (seven) is specifically as follows: locking arranged algae cells by utilizing 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 drag force of the fluid applied to the algae cells locked by the light spots is required to be smaller than the photoinduced dielectrophoresis force applied to the algae cells locked by the light spots, so that the deionized water washes and clears the rest algae cells which are not locked by the light spots from the micro-pipeline, and then the signal generator and the projector are turned off, so that the algae cells rotating in the micro-pipeline drive the fluid to flow from the fluid inlet to the fluid outlet.
The mechanism by which algae cells rotating within the micro-pipe in step (seven) drive fluid flow from the fluid inlet to the fluid outlet in the micro-pipe is: because the gap between the autorotation algae cells and the pipeline wall at the rear side in the micro-pipeline is different from the gap between the autorotation algae cells and the pipeline wall at the front side in the micro-pipeline, the viscous resistances of the fluid received by the algae cells are different in the rotation process of the algae cells, so that the algae cells generate a net flow to the fluid when rotating, and the fluid is driven to flow from the left end to the right end of the micro-pipeline: a row of counter-clockwise rotating algae cells near the inner rear pipeline wall of the micro-pipeline drives fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, a row of clockwise rotating algae cells near the inner front pipeline wall of the micro-pipeline also drives fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, and the driving effect produced by the algae cells rotating in opposite directions on two sides is consistent.
Compared with the prior art, the invention has outstanding substantive characteristics and remarkable progress, in particular, the invention takes the rotary algae cells as functional components in the traditional micropump, thus not only avoiding the difficulty of manufacturing a precise micro-actuator, but also solving the problem of energy supply, utilizing the energy of biological cells to complete the driving of fluid, without providing energy externally, and utilizing a plurality of algae cells to work cooperatively, thereby further improving the performance of the whole viscous micropump.
Drawings
Fig. 1 is a schematic structural view of the viscous micropump of the present invention.
Fig. 2 is a schematic structural view of the L-shaped interposer of the present invention.
Fig. 3 is a schematic structural view of a fluid driving chip of the present invention.
Fig. 4 is a schematic structural view of the microstructured layer of the present invention.
FIG. 5 is a schematic view of the structure of the lower ITO glass layer of the present invention.
FIG. 6 is a graph showing the distribution of algal cells of the present invention in a microchannel of a fluid driven chip.
FIG. 7 is a schematic diagram of a fluid driven chip of the present inventionFinite element simulation results.
FIG. 8 is a finite element simulation of algae cell driving fluid rotating in opposite directions on both sides of a microchannel in accordance with the present invention.
Detailed Description
Embodiments of the present invention are further described below with reference to the accompanying drawings.
As shown in fig. 1 to 6, a viscous micro pump is designed, the viscous micro pump comprises 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-stage 6, a projector 7, a focusing objective 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 left and right, a micro pipeline 11 between the fluid inlet and the fluid outlet is arranged in the fluid driving chip 1 along the left and right directions, the fluid inlet is connected with the left end of the micro pipeline 11, the fluid outlet is connected with the right end of the micro pipeline 11, the L-shaped adapter plate 2 comprises a horizontal plate and a vertical plate, the left side of the horizontal plate of the L-shaped adapter plate 2 is integrally and fixedly connected with the lower side of the vertical plate, the left side surface of the vertical plate of the L-shaped adapter plate 2 is fixedly connected to the three-dimensional nano moving platform 3, the middle part of the right side of the 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 being open and with a large upper part and a small lower part, the size of the fluid driving chip 1 is the same as that of the upper groove of the stepped rectangular groove 12, the fluid driving chip 1 is embedded in the upper groove of the stepped rectangular groove 12 in a matching way, the microscope 4 is arranged right above the fluid driving chip 1, the CCD5 is fixedly arranged on the upper part of the microscope 4, the lens barrel of the microscope 4 is fixedly connected with the high-precision Z-direction displacement platform 6, the projector 7 is arranged right below the fluid driving chip 1, the lens of the projector 7 is arranged at the top of the projector 7 and irradiates upwards, the focusing objective 8 is arranged at the upper part of the lens of the projector 7, the light path of the focusing objective 8 coincides with the light path of the microscope 4, the signal generator 9 is electrically connected with the fluid driving chip 1, and the computer 10 is respectively connected with the CCD5 and the projector 7 in a signal manner;
secondly, slowly injecting the treated algae cell solution into the micro-pipeline 11 in the fluid driving chip 1 from the fluid inlet;
drawing a plurality of spot images by using flash drawing software on a computer 10, wherein each spot image is projected upwards through a projector 7, converged by a focusing objective lens 8 and then projected onto the lower surface of the fluid driving chip 1 through the gap of a stepped rectangular groove 12;
fourthly, adjusting the three-dimensional nano-moving platform 3 and the high-precision Z-direction moving platform 6 to ensure that the focal plane of the focusing objective lens 8 and the focal plane of the microscope 4 are focused at the micro-pipeline 11 inside the fluid driving chip 1;
fifthly, adjusting the size of the light spot drawn in the step (III), adjusting the size of the light spot finally projected onto the fluid driving chip 1 to be 10-15 mu m, starting the signal generator 9, generating a sinusoidal signal with the frequency of 60KHz and the peak-to-peak value of 6V by the signal generator 9, and acting on the fluid driving chip 1 through a lead;
capturing the autorotation algae cells in the micro-pipeline 11 by utilizing the projected light spots and moving the algae cells to be arranged;
(seventh), removing the redundant algae cells from the micro-pipeline 11, and turning off the signal generator 9 and the projector 7, so that the algae cells rotating in the micro-pipeline 11 drive the fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline 11.
The fluid driving chip 1 comprises an upper ITO glass layer 13 and a lower ITO glass layer 14, a micro-structure 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 all along the left-right direction, the upper ITO glass layer 13 and the lower ITO glass layer 14 are rectangular sheet structures with the length of 25mm and the width of 20mm, the thicknesses of the upper ITO glass layer 13 and the lower ITO glass layer 14 are 1mm, two first through holes 16 with the diameter of 0.5mm and penetrating up and down are arranged on the upper ITO glass layer 13 at intervals left and right, the first through holes 16 on the left side are fluid inlets, the first through holes 16 on the right side are fluid outlets, the micro-structure layer 15 is of square sheet structure with the side length of 20mm, the thickness of the micro-structure layer 15 is 0.02-0.1mm, rectangular long holes 17 with the length of 10mm, the width of 0.1mm and penetrating up and down are arranged in the middle of the micro-structure layer 15 along the left-right direction, two second through holes 18 with the diameter of 0.5mm and penetrating up and down are arranged on the micro-structural layer 15 at intervals left and right, a 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 respectively and electrically connected with the two conductive surfaces through wires, the conductive surface of the lower ITO glass layer 14 is coated with a hydrogenated amorphous silicon layer 19 in a spin 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 micro-structural 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 micro-structural 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 aligned up and down respectively with the two second through holes 18, and the rectangular long holes 17 are enclosed into micro-channels 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 L-shaped adapter plate 2 is an aluminum plate, the width of the L-shaped adapter plate 2 is 50mm, the thickness of the L-shaped adapter plate 2 is 2mm, the height of a vertical plate of the L-shaped adapter plate 2 is 50mm, the length of a horizontal plate of the L-shaped adapter plate 2 is 200mm, the upper groove size of the stepped rectangular groove 12 is 25mm long, 20mm wide and 0.5mm thick, and the lower groove size of the stepped rectangular groove 12 is 24mm long, 18mm wide and 1.5mm thick.
The second step is specifically as follows: taking 1ml of algae cell solution, centrifuging at 3000rpm in a centrifuge for 6min, removing supernatant, adding deionized water again, and adjusting the concentration of the centrifuged algae cell solution to 1×10 5 cells/ml, then the treated algal cell solution is injected from the fluid inlet into the micro-channels 11 within the fluid driving chip 1.
The reason why the spot size finally projected onto the fluid driving chip 1 is adjusted to 10-15 μm in the step (five) is that: the shape of the algae cells is elliptical or oval, the size is about 10 mu m, and the light spot size is adjusted to be equivalent to the algae cell size, so that the capturing operation of single algae cells is more convenient to realize;
in the step (five), the sinusoidal signals generated by the signal generator 9 are respectively applied to the conductive surface of the upper ITO glass layer 13 and the conductive surface of the lower ITO glass layer 14 through wires.
The step (six) is specifically as follows: the CCD5 feeds back the image in the fluid driving chip 1 observed by the microscope 4 to the computer 10, and the flash drawing software on the computer 10 draws each light spot in combination with the fed back image, so as to move the light spot projected on the fluid driving chip 1, each light spot captures the algae cells rotating in the micro-pipeline 11 respectively, each clockwise rotated algae cell is arranged at a position close to the pipeline wall at the front side in the micro-pipeline 11 at a left-right interval, and each anticlockwise rotated algae cell is arranged at a position close to the pipeline wall at the rear side in the micro-pipeline 11 at a left-right interval.
The algae cell rotation mechanism 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 bending swing to a certain extent, the bending swing of the two flagella of the algae cell is generated by virtue of relative sliding caused by the conformational change of the dynamic protein between adjacent microtubules, when the two flagella are in a free state, the algae cell shows a random swimming movement form, and when only one flagellum is in the free swing, the algae cell shows a counterclockwise rotation or clockwise rotation movement 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 14, the conductivity of the hydrogenated amorphous silicon layer 19 is very low, at this time, 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 irradiates the hydrogenated amorphous silicon layer 19, the conductivity of the hydrogenated amorphous silicon layer 19 at the light spot area is sharply increased due to the electron hole pair generated by the light spot, the hydrogenated amorphous silicon layer 19 at the light spot area corresponds to a conductor, most of the voltage falls on the microstructure layer 15, so that an uneven electric field is generated in the microstructure layer 15 with the light spot area and the no light spot area, the algae cells are polarized in the uneven electric field area, and then are subjected to light induced dielectrophoresis as shown in the following expression:
in the middle ofIs the dielectric constant of the solution medium, +.>Is the diameter of algae cells, < > and->Is C-M factorReal part of->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 wires, the algal cells are subjected to a light-induced dielectrophoresis force directed to the spot area by the spot-free area, and the light-induced dielectrophoresis force is greater than the swimming force of the algal cells themselves, so that the spots can capture and move the algal cells.
The step (seven) is specifically as follows: the arranged algae cells are locked by utilizing each light spot, deionized water is injected into the micro-pipeline 11 from the fluid inlet at a certain flow rate, and the flow rate of the deionized water meets the following conditions: the drag force of the fluid applied to the algae cells locked by the light spots is required to be smaller than the photoinduced dielectrophoresis force applied to the algae cells locked by the light spots, so that the deionized water washes and clears the rest algae cells not locked by the light spots from the micro-pipeline 11, then the signal generator 9 and the projector 7 are turned off, and the algae cells rotating in the micro-pipeline 11 drive the fluid to flow from the fluid inlet to the fluid outlet.
The mechanism by which algae cells rotating within the micro-pipe 11 in step (seven) drive fluid flow from the fluid inlet to the fluid outlet in the micro-pipe 11 is: because the gap between the autorotation algae cells and the inner rear side pipeline wall of the micro pipeline 11 and the gap between the autorotation algae cells and the inner front side pipeline wall of the micro pipeline 11 are different, the viscous resistance of fluid borne by the algae cells in the rotation process of the algae cells is different, so that when the algae cells rotate, a net flow is generated for the fluid, and the fluid is driven to flow from the left end of the micro pipeline 11 to the right end: a row of counter-clockwise rotating algae cells near the inner rear wall of the microchannel 11 will drive fluid to flow from the fluid inlet to the fluid outlet in the microchannel 11, a row of clockwise rotating algae cells near the inner front wall of the microchannel 11 will also drive fluid to flow from the fluid inlet to the fluid outlet in the microchannel 11, and the driving effect of the counter-rotating algae cells on both sides will be consistent.
FIG. 7 shows the fluid driving chip 1 when a sinusoidal signal of 6V peak-to-peak value of 60KHz 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. Mu.mFinite element simulation results, the amplitude and direction of which represent the magnitude and direction of the photoinduced dielectrophoresis force, black arrows in the simulation results indicate +.>From the simulation results, it can be seen that most of the directions of the arrows are directed from the speckle-free region to the speckle-free region.
FIG. 8 shows the results of finite element simulation of the fluid driven by algae cells rotating counterclockwise at 6.28rad/s near the inner rear side wall of the microchannel 11 and algae cells rotating clockwise at 6.28rad/s near the inner front side wall of the microchannel 11 in the fluid driving chip 1, and it can be seen that the driving effect produced by algae cells rotating in opposite directions on both sides is consistent.
The above embodiments are merely for illustrating the technical aspects of the present invention, and it should be understood by those skilled in the art that the present invention is described in detail with reference to the above embodiments; modifications and equivalents may be made thereto without departing from the spirit and scope of the invention, which is intended to be encompassed by the claims.
Claims (8)
1. A method of algae cell-based fluid driving, characterized by: the method specifically comprises the following steps:
the method comprises the steps of (a) designing a viscous micropump, wherein the viscous micropump comprises a fluid driving chip, an L-shaped adapter plate, a three-dimensional nano movable 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;
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 by a focusing objective lens and then projected onto the lower surface of the fluid driving chip through a gap of a stepped rectangular groove upwards;
fourthly, adjusting the three-dimensional nano moving platform and the high-precision Z-direction moving platform to ensure that the focal plane of the focusing objective lens and the focal plane of the microscope are focused at the micro-channel inside the fluid driving chip;
fifthly, adjusting the size of the light spot drawn in the step (III), adjusting the size of the light spot finally projected onto the fluid driving chip to be 10-15 mu m, starting a signal generator, generating a sinusoidal signal with the frequency of 60KHz and the peak-to-peak value of 6V by the signal generator, and acting on the fluid driving chip through a lead;
capturing algae cells rotating in the micro-pipeline by utilizing the projected light spots and moving the algae cells to be arranged;
(seventh), removing redundant algae cells from the micro-pipeline, and turning off the signal generator and the projector, so that the algae cells rotating in the micro-pipeline drive fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline;
the fluid driving chip comprises an upper ITO glass layer and a lower ITO glass layer, a micro-structure 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 all arranged along the left-right direction, the upper ITO glass layer and the lower ITO glass layer are rectangular sheet structures 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 penetrating up and down are arranged on the upper ITO glass layer at intervals left and right, the left first through hole is a fluid inlet, the right first through hole is a fluid outlet, the micro-structure layer is a square sheet structure with the side length of 20mm, the thickness of the micro-structure layer is 0.02-0.1mm, a rectangular long hole with the length of 10mm and the width of 0.1mm and penetrating up and down is arranged in the left-right direction in the middle of the micro-structure layer, two second through holes with the diameter of 0.5mm and penetrating up and down are arranged on the micro-structure 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 left second through hole, the right end of the rectangular long hole is communicated with the right second through hole, 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 wires, the conductive surface of the lower ITO glass layer is spin-coated with a hydrogenated amorphous silicon 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 flush 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 silicon layer is flush with the lower surface of the microstructure layer, the microstructure layer and the hydrogenated amorphous silicon layer are vertically aligned, the two first through holes are respectively aligned with the two second through holes vertically, and the upper ITO glass layer, the micro-structure layer and the lower ITO glass layer are bonded to form a rectangular long hole which is a micro-pipe with the length of 10mm, the width of 0.1mm and the height of 0.02-0.1 mm;
the L-shaped adapter plate is an aluminum plate, the width of the L-shaped adapter plate is 50mm, the thickness of the L-shaped adapter plate is 2mm, the height of a vertical plate of the L-shaped adapter plate is 50mm, the length of a horizontal plate of the L-shaped adapter plate is 200mm, the upper groove size of the stepped rectangular groove is 25mm long, 20mm wide and 0.5mm thick, and the lower groove size of the stepped rectangular groove is 24mm long, 18mm wide and 1.5mm thick.
2. The algae cell-based fluid driving method of claim 1, wherein: the second step is specifically as follows: taking 1ml of algae cell solution, centrifuging at 3000rpm in a centrifuge for 6min, removing supernatant, adding deionized water again, and adjusting the concentration of the centrifuged algae cell solution to 1×10 5 cells/ml, then the treated algae cell solution is injected from the fluid inlet into the micro-tubing within the fluid-driven chip.
3. The algae cell-based fluid driving method of claim 2, wherein: the reason why the spot size finally projected onto the fluid driving chip is adjusted to 10-15 μm in the step (five) is that: the shape of the algae cells is elliptical or oval, the size is 10 mu m, and the light spot size is adjusted to be equivalent to the size of the algae cells, so that the capturing operation of single algae cells is more convenient to realize;
and (V) the sinusoidal signals generated by the signal generator in the step (V) are respectively acted on the conductive surface of the upper ITO glass layer and the conductive surface of the lower ITO glass layer through wires.
4. The algae cell-based fluid driving method of claim 3, wherein: the step (six) is specifically as follows: the image in the fluid driving chip observed by the microscope is fed back to the computer through the CCD, each light spot drawn by flash drawing software on the computer is moved in combination with the fed back image, then the light spot projected onto the fluid driving chip is moved, each light spot respectively captures algae cells rotating in the micro-pipeline, each clockwise rotated algae cell is arranged at a position close to the pipeline wall at the front side in the micro-pipeline at a left-right interval, and each anticlockwise rotated algae cell is arranged at a position close to the pipeline wall at the rear side in the micro-pipeline at a left-right interval.
5. The algae cell based fluid driving method of claim 4, wherein: the algae cell rotation mechanism 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 bending swing to a certain extent, the bending swing of the two flagella of the algae cell is generated by virtue of relative sliding caused by the conformational change of the dynamic protein between adjacent microtubules, when the two flagella are in a free state, the algae cell shows a random swimming movement form, and when only one flagellum is in the free swing, the algae cell shows a counterclockwise rotation or clockwise rotation movement form.
6. The algae cell based fluid driving method of claim 5, 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, at this time, 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 at the light spot area is sharply increased due to electron hole pairs generated by the light spot, the hydrogenated amorphous silicon layer at the light spot area is equivalent to a conductor, most of the voltage falls on the microstructure layer, thus a non-uniform electric field is generated in the microstructure layer with and without the light spot area, alga cells are polarized in the non-uniform electric field area and then receive photoinduction dielectrophoresis force, and the expression of the photoinduction force received by the alga cells is as follows:
in the middle ofIs the dielectric constant of the solution medium, +.>Is the diameter of algae cells, < > and->Is C-M factor->Real part of->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 wires, the algae cells are subjected to light-induced dielectrophoresis force with the spot-free area pointing to the spot area, and the light-induced dielectrophoresis force is larger than the swimming force of the algae cells, so that the light spots can capture and move the algae cells.
7. The algae cell based fluid driving method of claim 6, wherein: the step (seven) is specifically as follows: locking arranged algae cells by utilizing 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 drag force of the fluid applied to the algae cells locked by the light spots is required to be smaller than the photoinduced dielectrophoresis force applied to the algae cells locked by the light spots, so that the deionized water washes and clears the rest algae cells which are not locked by the light spots from the micro-pipeline, and then the signal generator and the projector are turned off, so that the algae cells rotating in the micro-pipeline drive the fluid to flow from the fluid inlet to the fluid outlet.
8. The algae cell-based fluid driving method of claim 7, wherein: the mechanism by which algae cells rotating within the micro-pipe in step (seven) drive fluid flow from the fluid inlet to the fluid outlet in the micro-pipe is: because the gap between the autorotation algae cells and the pipeline wall at the rear side in the micro-pipeline is different from the gap between the autorotation algae cells and the pipeline wall at the front side in the micro-pipeline, the viscous resistances of the fluid received by the algae cells are different in the rotation process of the algae cells, so that the algae cells generate a net flow to the fluid when rotating, and the fluid is driven to flow from the left end to the right end of the micro-pipeline: a row of counter-clockwise rotating algae cells near the inner rear pipeline wall of the micro-pipeline drives fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, a row of clockwise rotating algae cells near the inner front pipeline wall of the micro-pipeline also drives fluid to flow from the fluid inlet to the fluid outlet in the micro-pipeline, and the driving effect produced by the algae cells rotating in opposite directions on two sides is consistent.
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