CN210198937U - Unicellular adhesion measuring device based on scanning electron microscope - Google Patents
Unicellular adhesion measuring device based on scanning electron microscope Download PDFInfo
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- CN210198937U CN210198937U CN201920704807.1U CN201920704807U CN210198937U CN 210198937 U CN210198937 U CN 210198937U CN 201920704807 U CN201920704807 U CN 201920704807U CN 210198937 U CN210198937 U CN 210198937U
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Abstract
The utility model relates to the field of bioscience, a unicellular adhesion force measuring device based on scanning electron microscope, including the scanning electron microscope chamber, the gas holder, the vapor source, the intake pipe, real empty room, the electron gun, the electron detector, displacement platform I, the actuator, the micromanipulator, displacement platform II, the substrate, the cell sample, a computer, the scanning electron microscope, trachea and cable, displacement platform II is located 16 centimeters below the electron gun, the upper portion of substrate is parallel arrangement's strip arch, the electron detector is located displacement platform II's side top, the micromanipulator has front end and rear end, the front end has parallel arrangement's three control finger, the three-dimensional removal of displacement platform I and displacement platform II can be controlled to the computer, based on the scanning electron microscope, adopt special micromanipulator and substrate, can carry out adhesion force measurement in real time when handling the cell, because the micromanipulator picks up the cell from the downside of cell, the cell deformation is small in the measurement process, the experimental method is simple, and the accuracy of the experimental result is high.
Description
Technical Field
The utility model belongs to the technical field of biological science and specifically relates to an adopt single cell adhesion measuring device based on scanning electron microscope of special micromanipulator and substrate.
Background
The adhesion between cells and materials is an important characteristic of the cells, has important significance on the culture and regeneration of the cells, is generally qualitatively researched by measuring the adhesion between the cells and a substrate made of a certain material, and the adhesion of the cells is generally researched by a method for manipulating the cells by adopting devices such as an atomic force microscope, a micropipette and the like in the prior art.
SUMMERY OF THE UTILITY MODEL
In order to solve the problem, the utility model discloses the device is based on scanning electron microscope to adopt special micromanipulator and substrate, carry out adhesion in real time and measure when can handling the cell, the deformation of cell is little in the measurement process.
The utility model adopts the technical proposal that:
the single cell adhesion force measuring device based on the scanning electron microscope comprises a scanning electron microscope cavity, an air storage tank, a water vapor source, an air inlet pipe, a vacuum chamber, an electron gun, an electron detector, a displacement table I, an actuator, a micromanipulator, a displacement table II, a substrate, a cell sample, a computer, a scanning electron microscope, an air pipe and a cable, wherein xyz is a three-dimensional coordinate system, one end of the air inlet pipe is positioned in the scanning electron microscope cavity, the other end of the air inlet pipe is positioned outside the scanning electron microscope cavity and is respectively connected with the air storage tank and the water vapor source through the air pipe, the vacuum chamber is arranged above the scanning electron microscope cavity, the electron gun is connected with the lower end of the vacuum chamber, the electron gun, the electron detector, the displacement table I, the actuator, the micromanipulator, the displacement table II, the substrate and the cell sample are all positioned in the scanning electron microscope cavity, the electron detector, the displacement table I, the, the substrate is arranged on the displacement table II, the cell sample is adsorbed on the upper surface of the substrate, and the upper part of the substrate consists of parallel strip-shaped bulges and gaps; when the electron gun emits electrons to the substrate and interacts with the surface of the substrate, one part of the electrons forms reflected electrons, the electron detector is positioned above the side of the displacement table II and used for detecting the reflected electrons on the surface of the substrate, the electron detector can input detected electronic information into a computer, and the three-dimensional movement of the displacement table I and the displacement table II can be controlled through the computer; the micromanipulator is formed by processing a metal sheet with the length of 200 micrometers, the width of 40 micrometers and the thickness of 6 micrometers, and has elasticity, the micromanipulator is provided with a front end and a rear end, the front end is provided with three manipulating fingers which are arranged in parallel, the manipulating fingers are provided with a front section and a rear section which form an angle of 20 degrees with each other, the rear end of the micromanipulator is connected to the displacement table I through an actuator, a computer can control the actuator and drive the micromanipulator to rotate in an xz plane by taking the rear end of the micromanipulator as an axis, and the rotation range is plus or minus 15 degrees; the elastic coefficient of the micromanipulator is 0.7N/m, the three manipulation fingers at the front end of the micromanipulator are all 30 micrometers in length, 0.8 micrometer in width and 4 micrometers in height, the interval between the adjacent manipulation fingers is 1.2 micrometers, the length of the front section of the manipulation finger is 8 micrometers, the width is 0.8 micrometer and the height is 1 micrometer; the width of each strip-shaped bulge on the upper part of the substrate is 1 micron, the height of each strip-shaped bulge is 6 microns, the width of a gap between every two adjacent strip-shaped bulges is 1 micron, the substrate is formed by micromachining a polymethyl methacrylate (PMMA) sheet, and the surface of the substrate is plated with a gold film with the thickness of 20 microns; the size of the cells in the cell sample ranges from 2 microns to 4 microns, and the cell sample contains cells of different types;
the method for measuring the cell adhesion by using the single cell adhesion measuring device based on the scanning electron microscope comprises the following steps:
monitoring the relative position of a micromanipulator and a substrate by adopting a scanning electron microscope, and controlling the three-dimensional movement of a displacement table I and a displacement table II in the xyz direction by a computer so as to enable the micromanipulator and the substrate to approach each other;
secondly, obtaining images of the substrate and the cell sample through a scanning electron microscope, selecting a cell on the upper surface of the substrate as a cell to be detected to perform a subsequent measurement experiment, and controlling the movement of the displacement table II through a computer to enable the operating finger of the micromanipulator to move to a position with a distance of 100 microns above the side of the cell to be detected;
controlling an actuator by a computer and driving a micromanipulator to enable the front section of a manipulation finger of the micromanipulator to be parallel to a gap between the strip-shaped bulges on the upper part of the substrate where the cells to be detected are located;
controlling the displacement table II to move upwards through the computer, so that the operating finger of the micro-manipulator is embedded into a gap between the strip-shaped bulges where the cell to be detected is located;
controlling the displacement table II to move horizontally through the computer, so that the operating finger of the micro-manipulator moves along the gap between the strip-shaped bulges until the operating finger is positioned below the cell to be detected;
controlling the displacement table II to move downwards through the computer, enabling the front section of the manipulation finger of the micromanipulator to support the whole cell to be detected and separate the cell from the substrate, and monitoring the deformation of the micromanipulator through a scanning electron microscope to obtain the deformation distance delta of the micromanipulator in the vertical direction relative to the micromanipulator in the state of not being subjected to external force;
and seventhly, estimating the force F (k) delta applied to the micromanipulator, wherein k is the elastic coefficient of the micromanipulator, and finally obtaining the adhesive force of the cell to be detected when the cell is separated from the upper surface of the substrate.
The utility model has the advantages that:
the utility model discloses the adhesion can be carried out in real time when the device is handled the cell and the cell is measured, because the cell is picked up from the downside of cell to the micromanipulator, consequently the deformation of cell is little in the measurement process, and experimental method is simple, and measuring result is accurate.
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 an enlarged schematic view of the micromanipulator;
FIG. 3 is a top view of FIG. 2;
FIG. 4 is an enlarged schematic view of a substrate;
FIG. 5 is an enlarged view of one of the manipulation processes of the test cell by the micromanipulator;
FIG. 6 is an enlarged view of the second operation of the micromanipulator on the test cells;
FIG. 7 is an enlarged view of the third operation of the micromanipulator on the test cells;
FIG. 8 is an enlarged view of the fourth operation of the micromanipulator for manipulating the test cells.
In the figure, 1, a scanning electron microscope cavity, 2, an air storage tank, 3, a water vapor source, 4, an air inlet pipe, 5, a vacuum chamber, 6, an electron gun, 7, an electronic detector, 8, a displacement table I, 9, an actuator, 10, a micromanipulator, 11, a displacement table II, 12, a substrate, 13, a cell sample, 14 and a computer.
Detailed Description
FIG. 1 is a schematic diagram of the present invention, which comprises a scanning electron microscope chamber (1), a gas storage tank (2), a water vapor source (3), a gas inlet pipe (4), a vacuum chamber (5), an electron gun (6), an electron detector (7), a displacement table I (8), an actuator (9), a micromanipulator (10), a displacement table II (11), a substrate (12), a cell sample (13), a computer (14), a scanning electron microscope, a gas pipe and a cable, where xyz is a three-dimensional coordinate system, one end of the gas inlet pipe (4) is located in the scanning electron microscope chamber (1), the other end is located outside the scanning electron microscope chamber (1) and is connected with the gas storage tank (2) and the water vapor source (3) through the gas pipe, the vacuum chamber (5) is installed on the scanning electron microscope chamber (1), the electron gun (6) is connected to the lower end of the vacuum chamber (5), the electron gun (6), the electron detector (7), the displacement, The device comprises an actuator (9), a micromanipulator (10), a displacement table II (11), a substrate (12) and a cell sample (13), wherein the actuator (9), the micromanipulator (10), the displacement table II (11), the substrate (12) and the cell sample (13) are all located in a scanning electron mirror cavity (1), an electronic detector (7), a displacement table I (8), the actuator (9) and the displacement table II (11) are respectively connected with a computer (14) through cables, the displacement table II (11) is located at a 16-centimeter position below an electron gun (6), the substrate (12) is located on the displacement table II (11), the cell sample (13) is adsorbed on the upper surface of the substrate (12), and the upper part of the substrate (12) consists of; when the electron gun (6) emits electrons to the substrate (12) and interacts with the surface of the substrate (12), a part of the electrons forms reflected electrons, the electron detector (7) is positioned above the side of the displacement table II (11) and is used for detecting the reflected electrons on the surface of the substrate (12), the electronic detector (7) can input detected electronic information into the computer (14), and the three-dimensional movement of the displacement table I (8) and the displacement table II (11) can be controlled through the computer (14); the micromanipulator (10) is processed by a metal sheet with the length of 200 micrometers, the width of 40 micrometers and the thickness of 6 micrometers and has elasticity, the micromanipulator (10) is provided with a front end and a rear end, the front end is provided with three manipulating fingers which are arranged in parallel, the manipulating fingers are provided with a front section and a rear section which form an angle of 20 degrees with each other, the rear end of the micromanipulator (10) is connected to a displacement table I (8) through an actuator (9), a computer (14) can control the actuator (9) and drive the micromanipulator (10) to rotate in an xz plane by taking the rear end of the micromanipulator (10) as an axis, and the rotating range is plus or minus 15 degrees; the cells in the cell sample (13) range in size from 2 microns to 4 microns, with different types of cells being contained in the cell sample (13).
Fig. 2 is an enlarged schematic view of a micromanipulator, fig. 3 is a top view of fig. 2, the micromanipulator (10) is processed by a metal sheet with 200 micron length, 40 micron width and 6 micron thickness and has elasticity, the modulus of elasticity of the micromanipulator (10) is 0.7 n/m, the micromanipulator (10) has a front end and a rear end, the front end has three manipulating fingers arranged in parallel, the manipulating fingers have a front section and a rear section which form an angle of 20 degrees with each other, all three manipulating fingers at the front end of the micromanipulator (10) are 30 micron length, 0.8 micron width and 4 micron height, the interval between adjacent manipulating fingers is 1.2 micron, the front section of the manipulating fingers is 8 micron length, 0.8 micron width and 1 micron height.
As shown in fig. 4, which is an enlarged schematic view of a substrate, the upper part of the substrate (12) is composed of parallel strip-shaped protrusions and gaps, the width of each strip-shaped protrusion on the upper part of the substrate (12) is 1 micrometer, the height of each strip-shaped protrusion is 6 micrometers, the width of the gap between adjacent strip-shaped protrusions is 1 micrometer, the substrate (12) is formed by micromachining a polymethyl methacrylate (PMMA) sheet, and the surface of the substrate (12) is plated with a gold film with the thickness of 20 micrometers.
Referring to fig. 5, which is an enlarged schematic view of one of the cell manipulating processes of the micromanipulator, the computer (14) controls the actuator (9) and drives the micromanipulator (10) so that the front section of the manipulating finger of the micromanipulator (10) is parallel to the gap between the strip-shaped protrusions where the cell to be measured is located on the upper portion of the substrate (12).
Referring to fig. 6, which is an enlarged schematic view of the second process of manipulating the cells to be tested by the micromanipulator, the computer (14) controls the displacement table II (11) to move upward, so that the manipulation fingers of the micromanipulator (10) are embedded into the gaps between the strip-shaped protrusions where the cells to be tested are located.
Referring to fig. 7, which is an enlarged schematic view of the third process of manipulating the cell to be tested by the micro-manipulator, the computer (14) controls the displacement table II (11) to move horizontally, so that the manipulation finger of the micro-manipulator (10) moves along the gap between the strip-shaped protrusions until the manipulation finger is located under the cell to be tested.
Referring to fig. 8, which is an enlarged schematic view of the fourth operation process of the micromanipulator for the cell to be measured, the computer (14) controls the displacement table II (11) to move downward, so that the front section of the operation finger of the micromanipulator (10) holds up the whole cell to be measured and separates from the substrate (12).
Principle of scanning electron microscope: the scanning electron microscope is a common device in the prior art, an electron gun (6) and an electron detector (7) both belong to the scanning electron microscope, the working principle of the scanning electron microscope is the interaction between electrons and substances, namely, a focused high-energy electron beam is used for scanning the surface of a sample, secondary electrons emitted from the surface of the sample can reflect certain physical information on the sample, the secondary electrons are collected by the electron detector and converted into electric signals to be input into a computer, and the computer can obtain images related to the spatial characteristics of the sample after processing the electric signals.
Principle of adhesion measurement: the adhesion force of the cell to be tested on the substrate (12) is approximately equal to the force F applied to the micromanipulator (10), and the force applied to the micromanipulator (10) can be estimated by the formula F ═ k · δ, wherein k is the elastic coefficient of the micromanipulator (10), and δ is the deformation distance of the micromanipulator (10) in the vertical direction relative to the state that the micromanipulator is not applied with external force.
The single cell adhesion force measuring device based on the scanning electron microscope comprises a scanning electron microscope cavity (1), a gas storage tank (2), a water vapor source (3), a gas inlet pipe (4), a vacuum chamber (5), an electron gun (6), an electron detector (7), a displacement table I (8), an actuator (9), a micromanipulator (10), a displacement table II (11), a substrate (12), a cell sample (13), a computer (14), the scanning electron microscope, a gas pipe and a cable, wherein xyz is a three-dimensional coordinate system, one end of the gas inlet pipe (4) is positioned in the scanning electron microscope cavity (1), the other end of the gas inlet pipe is positioned outside the scanning electron microscope cavity (1) and is respectively connected with the gas storage tank (2) and the water vapor source (3) through the gas pipe, the vacuum chamber (5) is arranged on the scanning electron microscope cavity (1), the electron gun (6) is connected to the lower end of the vacuum chamber (5), the electron gun (6, The displacement table I (8), the actuator (9), the micromanipulator (10), the displacement table II (11), the substrate (12) and the cell sample (13) are all located in the scanning electron microscope cavity (1), the electronic detector (7), the displacement table I (8), the actuator (9) and the displacement table II (11) are respectively connected with the computer (14) through cables, the displacement table II (11) is located at a 16 cm position below the electron gun (6), the substrate (12) is located on the displacement table II (11), the cell sample (13) is adsorbed on the upper surface of the substrate (12), and the upper part of the substrate (12) is composed of parallel strip-shaped protrusions and gaps; when the electron gun (6) emits electrons to the substrate (12) and interacts with the surface of the substrate (12), a part of the electrons forms reflected electrons, the electron detector (7) is positioned above the side of the displacement table II (11) and is used for detecting the reflected electrons on the surface of the substrate (12), the electronic detector (7) can input detected electronic information into the computer (14), and the three-dimensional movement of the displacement table I (8) and the displacement table II (11) can be controlled through the computer (14); the micromanipulator (10) is processed by a metal sheet with the length of 200 micrometers, the width of 40 micrometers and the thickness of 6 micrometers and has elasticity, the micromanipulator (10) is provided with a front end and a rear end, the front end is provided with three manipulating fingers which are arranged in parallel, the manipulating fingers are provided with a front section and a rear section which form an angle of 20 degrees with each other, the rear end of the micromanipulator (10) is connected to a displacement table I (8) through an actuator (9), a computer (14) can control the actuator (9) and drive the micromanipulator (10) to rotate in an xz plane by taking the rear end of the micromanipulator (10) as an axis, and the rotating range is plus or minus 15 degrees; the elastic coefficient of the micromanipulator (10) is 0.7N/m, the three manipulation fingers at the front end of the micromanipulator (10) are all 30 micrometers in length, 0.8 micrometer in width and 4 micrometers in height, the interval between the adjacent manipulation fingers is 1.2 micrometers, the length of the front section of the manipulation finger is 8 micrometers, 0.8 micrometer in width and 1 micrometer in height; the width of each strip-shaped bulge on the upper part of the substrate (12) is 1 micron, the height of each strip-shaped bulge is 6 microns, the width of a gap between every two adjacent strip-shaped bulges is 1 micron, the substrate (12) is formed by micromachining a polymethyl methacrylate (PMMA) sheet, and the surface of the substrate (12) is plated with a gold film with the thickness of 20 microns; the cells in the cell sample (13) range in size from 2 microns to 4 microns, with different types of cells being contained in the cell sample (13).
The utility model discloses the device combines special micromanipulator and substrate on scanning electron microscope's basis, measures cell adhesion when handling the cell, and the deformation of cell is little in the measurement process, and the experimental result degree of accuracy is high.
Claims (4)
1. A unicellular adhesion force measuring device based on a scanning electron microscope comprises a scanning electron microscope cavity (1), a gas storage tank (2), a water vapor source (3), a gas inlet pipe (4), a vacuum chamber (5), an electron gun (6), an electron detector (7), a displacement table I (8), an actuator (9), a micromanipulator (10), a displacement table II (11), a substrate (12), a cell sample (13), a computer (14), the scanning electron microscope, a gas pipe and a cable, wherein xyz is a three-dimensional coordinate system, one end of the gas inlet pipe (4) is positioned in the scanning electron microscope cavity (1), the other end of the gas inlet pipe is positioned outside the scanning electron microscope cavity (1) and is respectively connected with the gas storage tank (2) and the water vapor source (3) through the gas pipe, the vacuum chamber (5) is arranged on the scanning electron microscope cavity (1), the electron gun (6) is connected to the lower end of the vacuum chamber (5), the electron gun (, The displacement table I (8), the actuator (9), the micromanipulator (10), the displacement table II (11), the substrate (12) and the cell sample (13) are all positioned in the scanning electron microscope cavity (1), the electronic detector (7), the displacement table I (8), the actuator (9) and the displacement table II (11) are respectively connected with a computer (14) through cables,
the method is characterized in that: the displacement platform II (11) is positioned at a 16 cm position below the electron gun (6), the substrate (12) is arranged on the displacement platform II (11), the cell sample (13) is adsorbed on the upper surface of the substrate (12), and the upper part of the substrate (12) consists of parallel strip-shaped bulges and gaps; when the electron gun (6) emits electrons to the substrate (12) and interacts with the surface of the substrate (12), a part of the electrons forms reflected electrons, the electron detector (7) is positioned above the side of the displacement table II (11) and is used for detecting the reflected electrons on the surface of the substrate (12), the electronic detector (7) can input detected electronic information into the computer (14), and the three-dimensional movement of the displacement table I (8) and the displacement table II (11) can be controlled through the computer (14); the micromanipulator (10) is formed by processing a metal sheet with the length of 200 micrometers, the width of 40 micrometers and the thickness of 6 micrometers, and is elastic, the micromanipulator (10) is provided with a front end and a rear end, the front end is provided with three manipulating fingers which are arranged in parallel, the manipulating fingers are provided with a front section and a rear section which form an angle of 20 degrees with each other, the rear end of the micromanipulator (10) is connected to a displacement table I (8) through an actuator (9), a computer (14) can control the actuator (9) and drive the micromanipulator (10) to rotate in an xz plane by taking the rear end of the micromanipulator (10) as an axis, and the rotating range is plus or minus 15 degrees.
2. The single-cell adhesion force measuring device based on the scanning electron microscope as claimed in claim 1, wherein: the elastic coefficient of the micromanipulator (10) is 0.7N/m, the three manipulating fingers at the front end of the micromanipulator (10) are all 30 micrometers in length, 0.8 micrometer in width and 4 micrometers in height, the interval between the adjacent manipulating fingers is 1.2 micrometers, and the front section of the manipulating fingers is 8 micrometers in length, 0.8 micrometer in width and 1 micrometer in height.
3. The single-cell adhesion force measuring device based on the scanning electron microscope as claimed in claim 1, wherein: the width of each strip-shaped bulge on the upper part of the substrate (12) is 1 micrometer, the height of each strip-shaped bulge is 6 micrometers, the width of a gap between every two adjacent strip-shaped bulges is 1 micrometer, the substrate (12) is formed by micromachining a polymethyl methacrylate (PMMA) sheet, and a gold film with the thickness of 20 micrometers is plated on the surface of the substrate (12).
4. The single-cell adhesion force measuring device based on the scanning electron microscope as claimed in claim 1, wherein: the cells in the cell sample (13) range in size from 2 microns to 4 microns, with different types of cells being contained in the cell sample (13).
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| CN201920704807.1U CN210198937U (en) | 2019-05-11 | 2019-05-11 | Unicellular adhesion measuring device based on scanning electron microscope |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110108634A (en) * | 2019-05-11 | 2019-08-09 | 金华职业技术学院 | A kind of unicellular adherency force measuring device based on scanning electron microscope |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110108634A (en) * | 2019-05-11 | 2019-08-09 | 金华职业技术学院 | A kind of unicellular adherency force measuring device based on scanning electron microscope |
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