CN210533870U - Micro-nano compression device - Google Patents
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- CN210533870U CN210533870U CN201921039689.3U CN201921039689U CN210533870U CN 210533870 U CN210533870 U CN 210533870U CN 201921039689 U CN201921039689 U CN 201921039689U CN 210533870 U CN210533870 U CN 210533870U
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Abstract
The utility model relates to the technical field of bioscience, a micro-nano compression device, which comprises an optical microscope, a glass substrate, a metal foil, a filling layer, a micro-compressor, a liquid inlet pipe, a liquid outlet, a protective layer, an electromagnet I, an electromagnet II, a voltage source and a cable, wherein a compression experimental material comprises a macromolecule ball, a macromolecule sample, a magnetic ball and liquid, the micro-compressor comprises a metal probe I, a metal probe II, a micro-channel I, a micro-channel II, a compression channel, a port I, a port II, a port III and a port IV, the optical microscope is positioned at a position of 10 cm below the glass substrate, a micro-fluid structure is combined with magnetic force to apply compression force to a biological target in a finite space, the sample position is relatively local in the compression process, the compression effect is good, and the environment applied by the pressure is similar to the original living environment of, and the compression process can be monitored by adopting the conventional commercial optical microscope, so that the sample cannot deteriorate.
Description
Technical Field
The utility model belongs to the technical field of biological science technique and specifically relates to a receive compressor arrangement a little for exerting compressive force and observing its reaction to the biological target in the finite space.
Background
In recent years, it becomes more and more important to study the response of biological cells to external pressure stimulation on a molecular scale, generally, the prior art adopts a contact probe technology such as a micro-pressing plate, a micro-nano indentation technology, an atomic force microscope and the like, and applies unidirectional force to a biological sample on a substrate, which has the disadvantage of causing the sample to move in an unpressurized direction and affecting a compression effect, and the other prior art adopts an optical trap to manipulate a small ball positioned in a microfluidic channel to apply a compression force to the sample, and uses laser to generate the force.
SUMMERY OF THE UTILITY MODEL
In order to solve the above problems, the present invention combines microfluidic control and magnetic force for applying a compressive force to a biological sample in a confined space, and can monitor the compression process using a commercial optical microscope.
The utility model adopts the technical proposal that:
the micro-nano compression device comprises an optical microscope, a glass substrate, a metal foil, a filling layer, a micro-compressor, a liquid inlet pipe, a liquid inlet, a liquid outlet pipe, a liquid outlet, a protective layer, an electromagnet I, an electromagnet II, a voltage source and a cable, wherein xyz is a three-dimensional coordinate system, a compression experiment material comprises a macromolecular pellet, a macromolecular sample, a magnetic pellet and liquid, and the micro-compressor comprises a metal probe I, a metal probe II, a microchannel I, a microchannel II, a compression channel, a port I, a port II, a port III and a port IV; the middle position of the upper surface of the glass substrate is connected with a micro-compressor, metal foils with the thickness of 500 micrometers are deposited at the two side positions of the upper surface of the glass substrate, the rest space with the height of 500 micrometers on the upper surface of the glass substrate is a filling layer, the filling layer completely covers the micro-compressor, a liquid inlet is connected with a port II of the micro-compressor through a liquid inlet pipe, a liquid outlet is connected with a port III of the micro-compressor through a liquid outlet pipe, an electromagnet I and an electromagnet II are respectively fixed on the upper surfaces of the two metal foils, a protective layer covers the filling layer, the liquid inlet, the liquid outlet and the protective layer covers the upper parts of the liquid inlet, the liquid; the micro-compressor comprises a silicon chip substrate and a micro-nano structure on the silicon chip substrate, wherein a micro-channel I, a micro-channel II and a compression channel are all micro-fluid channels, two ends of the micro-channel I are respectively provided with a port I and a port III, two ends of the micro-channel II are respectively provided with a port II and a port IV, the port I and the port IV are sealed, a plurality of compression channels which are parallel to each other are arranged between the micro-channel I and the micro-channel II, the interval between every two adjacent compression channels is 4 micrometers, two ends of each compression channel are respectively communicated with the micro-channel I and the micro-channel II, the depth of each compression channel is 4 micrometers, the width of each compression channel is suddenly changed from 4 micrometers to 2 micrometers according to the liquid flow z negative direction, the section with the width of 4 micrometers is 80 micrometers, and the; the metal probe I and the metal probe II are all three metal electrodes in a group, the tail end of each metal electrode is in a needle point shape, the distance between the metal probe I and the micro-channel I is 30 micrometers, and the distance between the metal probe II and the micro-channel II is 30 micrometers; the voltage source can respectively apply voltage to the metal probe I and the metal probe II through cables, the voltage source can respectively apply voltage to the electromagnet I and the electromagnet II through the cables to generate a magnetic field, the position of the magnetic field in a compression channel is a uniform magnetic field of magnetic force lines along the z direction, the magnetic field intensity is 4000 gauss, and the magnetic field covers a micro-compressor area; the macromolecule globule, the macromolecule sample and the magnetic globule can be respectively injected into the micro-channel II and the compression channel through the liquid inlet; the filling layer is made of siloxane material; the micro-channel I and the micro-channel II are both 1mm in length, 120 microns in width and 60 microns in depth and are both formed by micromachining polymethyl methacrylate materials; the compression channels are processed on the silicon chip substrate by a photoetching method, and the length of each compression channel is 100 micrometers; the thicknesses of the metal probe I and the metal probe II are both 120 micrometers, and the tip-shaped curvature radius of the tail end of each metal electrode is 100 micrometers; the diameter of the polymer small ball is 3 microns and is made of polystyrene material; the magnetic beads had a diameter of 3.5 μm and were made of a ferromagnetic material and a magnetic permeability of 0.01H/m.
The method for compressing the biomacromolecule by adopting the micro-nano compression device comprises the following steps:
step one, observing the condition in a compression channel in a micro compressor by adopting an optical microscope;
injecting liquid containing polymer pellets into the microchannel II from the liquid inlet, wherein each microliter of the liquid contains 10000 polymer pellets, and the flow rate of the liquid is 0.3 microliter/hour until most of the compression channels contain one polymer pellet;
injecting liquid containing the macromolecular sample from a liquid inlet, wherein the concentration of the macromolecular sample in the liquid is 0.01mM, the flow rate of the liquid is 0.1 microliter/hour, and the macromolecular sample is contained in 80 percent of the compression channels;
injecting liquid containing the magnetic small balls from a liquid inlet, wherein each microliter of the liquid contains 6000 magnetic small balls, the flow rate of the liquid is 0.1 microliter/hour, and the magnetic small balls are arranged in 80 percent of compression channels;
step five, closing the liquid inlet and the liquid outlet;
step six, a voltage source respectively applies voltage to the electromagnet I and the electromagnet II so as to enable the electromagnet I and the electromagnet II to generate magnetic fields, and the voltage source respectively applies voltage to the metal probe I and the metal probe II so as to finely adjust the magnetic field intensity of the region between the metal probe I and the metal probe II;
step seven, the magnetic small balls in the compression channel move to one side of the macromolecular small balls in the compression channel under the action of magnetic force, and meanwhile, the macromolecular samples in the compression channel are compressed;
and step eight, recording the compressed image characteristics of the macromolecular sample observed by the optical microscope, and analyzing.
The utility model has the advantages that:
the utility model discloses the device can exert compressive force to the biological sample in finite space, and the in-process sample position that receives the compression is local relatively, and the compression is effectual, and the environment that pressure was applyed is similar with biological sample's original living environment, can not cause the sample rotten.
Drawings
The following is further illustrated in connection with the figures of the present invention:
FIG. 1 is a schematic view of the present invention;
FIG. 2 is a schematic diagram of a top view of a micro-compressor at an enlarged scale;
FIG. 3 is an enlarged schematic view of a compression channel for compressing a bio-macromolecule.
In the figure, 1, an optical microscope, 2, a glass substrate, 3, a metal foil, 4, a filling layer, 5, a micro compressor, 5-1, a metal probe I, 5-2, a metal probe II, 5-3, a micro channel I, 5-4, a micro channel II, 5-5, a compression channel, 5-6, a port I, 5-7, a port II, 5-8, a port III, 5-9, a port IV, 6, a liquid inlet pipe, 7, a liquid inlet, 8, a liquid outlet pipe, 9, a liquid outlet, 10, a protective layer, 11, an electromagnet I, 12, an electromagnet II, 13, a polymer pellet, 14, a macromolecule sample and 15, a magnetic pellet are arranged.
Detailed Description
FIG. 1 is a schematic view of the present invention, which comprises an optical microscope (1), a glass substrate (2), a metal foil (3), a filling layer (4), a micro-compressor (5), a liquid inlet pipe (6), a liquid inlet (7), and a liquid outlet pipe (8), the device comprises a liquid outlet (9), a protective layer (10), an electromagnet I (11), an electromagnet II (12), a voltage source and a cable, wherein xyz is a three-dimensional coordinate system, compression experimental materials comprise a polymer bead (13), a macromolecule sample (14), a magnetic bead (15) and liquid, a micro compressor (5) is connected to the middle position of the upper surface of a glass substrate (2), metal foils (3) with the thickness of 500 micrometers are deposited on the two side positions of the upper surface of the glass substrate (2), a filling layer (4) is arranged in the rest space with the height of 500 micrometers on the upper surface of the glass substrate (2), the filling layer (4) completely covers the micro compressor (5), and the filling layer (4) is made of siloxane materials; liquid inlet (7) is connected with port II (5-7) of micro-compressor (5) through liquid inlet pipe (6), liquid outlet (9) is connected with port III (5-8) of micro-compressor (5) through liquid outlet pipe (8), electromagnet I (11) and electromagnet II (12) are fixed on metal foil (3) of both sides respectively, filling layer (4), liquid inlet pipe (6), liquid inlet (7), protective layer (10) covers above liquid outlet (9) and liquid outlet (8), optical microscope (1) is located 10 cm position below glass substrate (2) for observe micro-compressor (5).
FIG. 2 is a schematic top view enlarged view of a micro-compressor, wherein the micro-compressor (5) comprises a metal probe I (5-1), a metal probe II (5-2), a micro-channel I (5-3), a micro-channel II (5-4), a compression channel (5-5), a port I (5-6), a port II (5-7), a port III (5-8) and a port IV (5-9), the micro-compressor (5) comprises a silicon wafer substrate and a micro-nano structure thereon, the micro-channel I (5-3), the micro-channel II (5-4) and the compression channel (5-5) are all micro-fluid channels, the micro-channel I (5-3) and the micro-channel II (5-4) are all micro-machined from a polymethyl methacrylate material, and have a length of 1mm, a width of 120 micrometers and a depth of 60 micrometers, the two ends of the micro-channel I (5-3) are respectively provided with a port I (5-6) and a port III (5-8), the two ends of the micro-channel II (5-4) are respectively provided with a port II (5-7) and a port IV (5-9), the port I (5-6) and the port IV (5-9) are sealed, a plurality of compression channels (5-5) which are parallel to each other are arranged between the micro-channel I (5-3) and the micro-channel II (5-4), the interval between the adjacent compression channels (5-5) is 4 micrometers, the compression channels (5-5) are processed on a silicon wafer substrate by a photoetching method, the length of each compression channel (5-5) is 100 micrometers, the two ends of each compression channel (5-5) are respectively communicated with the micro-channel I (5-3) and the micro-channel II (5-4), the depth of each compression channel (5-5) is 4 micrometers, the width of each compression channel is changed from 4 micrometers to 2 micrometers according to the liquid flow z negative direction, the length of the section with the width of 4 micrometers is 80 micrometers, the length of the section with the width of 2 micrometers is 20 micrometers, the section with the width of 4 micrometers is communicated with the microchannel II (5-4), and the section with the width of 2 micrometers is communicated with the microchannel I (5-3); the metal probe I (5-1) and the metal probe II (5-2) are a group of three metal electrodes, the tail end of each metal electrode is in a needle point shape, the thicknesses of the metal probe I (5-1) and the metal probe II (5-2) are 120 micrometers, the curvature radius of the needle point shape at the tail end of each metal electrode is 100 micrometers, the distance between the metal probe I (5-1) and the microchannel I (5-3) is 30 micrometers, and the distance between the metal probe II (5-2) and the microchannel II (5-4) is 30 micrometers; the voltage source can respectively apply voltage to the metal probe I (5-1) and the metal probe II (5-2) through cables, the voltage source can respectively apply voltage to the electromagnet I (11) and the electromagnet II (12) through the cables to generate a magnetic field, the magnetic field is a uniform magnetic field with magnetic lines of force along the z direction at the position of the compression channel (5-5), the magnetic field strength is 4000 gauss, and the magnetic field covers the area of the micro compressor (5); the macromolecule beads (13), the macromolecule sample (14) and the magnetic beads (15) can be respectively injected into the micro-channel II (5-4) and the compression channel (5-5) through the liquid inlet (7).
As shown in FIG. 3, which is an enlarged schematic view of a compression channel in the process of compressing biological macromolecules, an optical microscope (1) can be used to observe the situation in the compression channel (5-5) in the micro-compressor (5), the polymer beads (13) have a diameter of 3 microns and are made of polystyrene material, the magnetic beads (15) have a diameter of 3.5 microns and are made of ferromagnetic material and have a magnetic permeability of 0.01H/m, liquid containing the polymer beads (13) is injected into the micro-channel II (5-4) from a liquid inlet (7) at a liquid flow rate of 0.3 microliter/hr, and because the ports I (5-6) and IV (5-9) are sealed, the liquid containing the polymer beads (13) flows through the micro-channel II (5-4), the compression channel (5-5), the micro-channel I (5-3), After the polymer small balls (13) enter one compression channel (5-5) along with the flowing of liquid, the polymer small balls are blocked by a section of the compression channel (5-5) connected with the micro channel I (5-3) so as to be trapped in the compression channel (5-5), the flow rate of the liquid in the compression channel (5-5) is reduced, and therefore other polymer small balls (13) in the liquid are difficult to enter the compression channel (5-5); injecting a liquid containing the macromolecular sample (14) from the liquid inlet (7) at a flow rate of 0.1 microliter/hour after the macromolecular beads (13) are arranged in most of the compression channels (5-5), so that most of the compression channels (5-5) are provided with the macromolecular sample (14); injecting a liquid containing the magnetic beads (15) from the liquid inlet (7) at a flow rate of 0.1 microliter/hour until most of the compression passages (5-5) have the magnetic beads (15), closing the liquid inlet (7) and the liquid outlet (9); a voltage source respectively applies voltage to an electromagnet I (11) and an electromagnet II (12) to enable the electromagnet I (11) and the electromagnet II (12) to generate magnetic fields, the magnetic fields cover the area of the micro compressor (5), the needle point-shaped structures at the tail ends of the metal electrodes in the metal probe I (5-1) and the metal probe II can enable the nearby area to generate higher magnetic field gradient, the voltage source respectively applies voltage to the metal probe I (5-1) and the metal probe II (5-2) to enable the magnetic field intensity of the area between the metal probe I (5-1) and the metal probe II to be finely adjusted, so that the magnetic small balls (15) in the compression channel (5-5) are affected by magnetic force to move to one side of the polymer small balls (13) in the compression channel (5-5), and meanwhile, the macromolecule samples (14) in the compression channel (5-5) are compressed, there may be two or more magnetic beads (15) in some of the compression channels (5-5), and therefore the macromolecular samples (14) in these compression channels (5-5) are subjected to greater compressive forces under the same magnetic field conditions.
The micro-nano compression device comprises an optical microscope (1), a glass substrate (2), a metal foil (3), a filling layer (4), a micro-compressor (5), a liquid inlet pipe (6), a liquid inlet (7), a liquid outlet pipe (8), a liquid outlet (9), a protective layer (10), an electromagnet I (11), an electromagnet II (12), a voltage source and a cable, xyz is a three-dimensional coordinate system, the compression experimental material comprises a macromolecule ball (13), a macromolecule sample (14), a magnetic ball (15) and liquid, the micro compressor (5) comprises a metal probe I (5-1), a metal probe II (5-2), a micro channel I (5-3), a micro channel II (5-4), a compression channel (5-5), a port I (5-6), a port II (5-7), a port III (5-8) and a port IV (5-9); a micro-compressor (5) is connected to the middle position on the glass substrate (2), metal foils (3) with the thickness of 500 micrometers are deposited on the two side positions, a filling layer (4) is arranged in the rest space with the height of 500 micrometers on the glass substrate (2), the filling layer (4) completely covers the micro-compressor (5), a liquid inlet (7) is connected with ports II (5-7) of the micro-compressor (5) through a liquid inlet pipe (6), a liquid outlet (9) is connected with ports III (5-8) of the micro-compressor (5) through a liquid outlet pipe (8), electromagnets I (11) and II (12) are respectively fixed on the two metal foils (3), a protective layer (10) covers the filling layer (4), the liquid inlet (6), the liquid inlet (7), the liquid outlet pipe (8) and the liquid outlet (9), an optical microscope (1) is located at the 10 cm position below the glass substrate (2), for observing the micro-compressor (5); the micro compressor (5) consists of a silicon chip substrate and a micro-nano structure on the silicon chip substrate, the micro channel I (5-3), the micro channel II (5-4) and the compression channel (5-5) are all micro fluid channels, two ends of the micro channel I (5-3) are respectively provided with a port I (5-6) and a port III (5-8), two ends of the micro channel II (5-4) are respectively provided with a port II (5-7) and a port IV (5-9), the port I (5-6) and the port IV (5-9) are sealed, a plurality of compression channels (5-5) which are parallel to each other are arranged between the micro channel I (5-3) and the micro channel II (5-4), the interval between the adjacent compression channels (5-5) is 4 microns, two ends of each compression channel (5-5) are respectively communicated with the micro channel I (5-3) and the micro channel II (5-4), the depth of each compression channel (5-5) is 4 micrometers, the width of each compression channel is changed from 4 micrometers to 2 micrometers according to the negative direction of the liquid flow z, the length of the section with the width of 4 micrometers is 80 micrometers, and the length of the section with the width of 2 micrometers is 20 micrometers; the metal probe I (5-1) and the metal probe II (5-2) are all a group of three metal electrodes, the tail end of each metal electrode is in a needle point shape, the distance between the metal probe I (5-1) and the micro-channel I (5-3) is 30 micrometers, and the distance between the metal probe II (5-2) and the micro-channel II (5-4) is 30 micrometers; the voltage source can respectively apply voltage to the metal probe I (5-1) and the metal probe II (5-2) through cables, the voltage source can respectively apply voltage to the electromagnet I (11) and the electromagnet II (12) through the cables to generate a magnetic field, the magnetic field is a uniform magnetic field with magnetic lines of force along the z direction at the position of the compression channel (5-5), the magnetic field strength is 4000 gauss, and the magnetic field covers the area of the micro compressor (5); the macromolecule globule (13), the macromolecule sample (14) and the magnetic globule (15) can be respectively injected into the micro-channel II (5-4) and the compression channel (5-5) through the liquid inlet (7); the filling layer (4) is a siloxane material; the micro-channel I (5-3) and the micro-channel II (5-4) are both 1mm in length, 120 microns in width and 60 microns in depth and are formed by micromachining polymethyl methacrylate materials; the compression channels (5-5) are processed on the silicon chip substrate by a photoetching method, and the length of each compression channel (5-5) is 100 micrometers; the thicknesses of the metal probe I (5-1) and the metal probe II (5-2) are both 120 micrometers, and the radius of curvature of the tip shape of the tail end of each metal electrode is 100 micrometers; the diameter of the polymer small ball (13) is 3 microns and is made of polystyrene material; the magnetic beads (15) have a diameter of 3.5 μm and are made of a ferromagnetic material and have a magnetic permeability of 0.01H/m.
The utility model discloses the device combines together microfluid structure and magnetic force to adopt current commercial optical microscope to monitor compression process, can exert compressive force to the biological sample in finite space, and the original living environment that the environment that pressure was exerted can simulate biological sample.
Claims (7)
1. A micro-nano compression device comprises an optical microscope (1), a glass substrate (2), a metal foil (3), a filling layer (4), a micro-compressor (5), a liquid inlet pipe (6), a liquid inlet (7), a liquid outlet pipe (8), a liquid outlet (9), a protective layer (10), an electromagnet I (11), an electromagnet II (12), a voltage source and a cable, wherein xyz is a three-dimensional coordinate system, compression experiment materials comprise a polymer bead (13), a polymer sample (14), a magnetic bead (15) and liquid, the micro-compressor (5) comprises a metal probe I (5-1), a metal probe II (5-2), a micro-channel I (5-3), a micro-channel II (5-4), a compression channel (5-5), a port I (5-6), a port II (5-7), a port III (5-8) and a port IV (5-9),
the method is characterized in that: a micro-compressor (5) is connected to the middle position on the glass substrate (2), metal foils (3) with the thickness of 500 micrometers are deposited on the two side positions, a filling layer (4) is arranged in the rest space with the height of 500 micrometers on the glass substrate (2), the filling layer (4) completely covers the micro-compressor (5), a liquid inlet (7) is connected with ports II (5-7) of the micro-compressor (5) through a liquid inlet pipe (6), a liquid outlet (9) is connected with ports III (5-8) of the micro-compressor (5) through a liquid outlet pipe (8), electromagnets I (11) and II (12) are respectively fixed on the two metal foils (3), a protective layer (10) covers the filling layer (4), the liquid inlet (6), the liquid inlet (7), the liquid outlet pipe (8) and the liquid outlet (9), an optical microscope (1) is located at the 10 cm position below the glass substrate (2), for observing the micro-compressor (5);
the micro compressor (5) consists of a silicon chip substrate and a micro-nano structure on the silicon chip substrate, the micro channel I (5-3), the micro channel II (5-4) and the compression channel (5-5) are all micro fluid channels, two ends of the micro channel I (5-3) are respectively provided with a port I (5-6) and a port III (5-8), two ends of the micro channel II (5-4) are respectively provided with a port II (5-7) and a port IV (5-9), the port I (5-6) and the port IV (5-9) are sealed, a plurality of compression channels (5-5) which are parallel to each other are arranged between the micro channel I (5-3) and the micro channel II (5-4), the interval between the adjacent compression channels (5-5) is 4 microns, two ends of each compression channel (5-5) are respectively communicated with the micro channel I (5-3) and the micro channel II (5-4), the depth of each compression channel (5-5) is 4 micrometers, the width of each compression channel is changed from 4 micrometers to 2 micrometers, the length of the section with the width of 4 micrometers is 80 micrometers, and the length of the section with the width of 2 micrometers is 20 micrometers; the metal probe I (5-1) and the metal probe II (5-2) are all a group of three metal electrodes, the tail end of each metal electrode is in a needle point shape, the distance between the metal probe I (5-1) and the micro-channel I (5-3) is 30 micrometers, and the distance between the metal probe II (5-2) and the micro-channel II (5-4) is 30 micrometers; the voltage source can respectively apply voltage to the metal probe I (5-1) and the metal probe II (5-2) through cables, the voltage source can respectively apply voltage to the electromagnet I (11) and the electromagnet II (12) through the cables to generate a magnetic field, the magnetic field is a uniform magnetic field with magnetic lines of force along the z direction at the position of the compression channel (5-5), the magnetic field strength is 4000 gauss, and the magnetic field covers the area of the micro compressor (5); the macromolecule beads (13), the macromolecule sample (14) and the magnetic beads (15) can be respectively injected into the micro-channel II (5-4) and the compression channel (5-5) through the liquid inlet (7).
2. A micro-nano compression device according to claim 1, which is characterized in that: the filling layer (4) is a siloxane material.
3. A micro-nano compression device according to claim 1, which is characterized in that: the micro-channel I (5-3) and the micro-channel II (5-4) are both 1mm in length, 120 microns in width and 60 microns in depth, and are both formed by micromachining polymethyl methacrylate materials.
4. A micro-nano compression device according to claim 1, which is characterized in that: the compression channels (5-5) are processed on the silicon chip substrate by a photoetching method, and the length of each compression channel (5-5) is 100 micrometers.
5. A micro-nano compression device according to claim 1, which is characterized in that: the thickness of each of the metal probe I (5-1) and the metal probe II (5-2) is 120 micrometers, and the radius of curvature of the tip shape of each metal electrode is 100 micrometers.
6. A micro-nano compression device according to claim 1, which is characterized in that: the polymer beads (13) have a diameter of 3 μm and are made of polystyrene material.
7. A micro-nano compression device according to claim 1, which is characterized in that: the magnetic beads (15) have a diameter of 3.5 μm and are made of a ferromagnetic material and have a magnetic permeability of 0.01H/m.
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| CN110308041A (en) * | 2019-06-28 | 2019-10-08 | 金华职业技术学院 | A kind of micro-nano compression set |
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| CN110308041A (en) * | 2019-06-28 | 2019-10-08 | 金华职业技术学院 | A kind of micro-nano compression set |
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