CN112834573A - A rapid test method for biological effects of medium frequency micro-electric field - Google Patents
A rapid test method for biological effects of medium frequency micro-electric field Download PDFInfo
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- CN112834573A CN112834573A CN202011626679.7A CN202011626679A CN112834573A CN 112834573 A CN112834573 A CN 112834573A CN 202011626679 A CN202011626679 A CN 202011626679A CN 112834573 A CN112834573 A CN 112834573A
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- 230000004071 biological effect Effects 0.000 title claims abstract description 32
- 238000010998 test method Methods 0.000 title claims abstract description 14
- 230000005684 electric field Effects 0.000 claims abstract description 94
- 238000012360 testing method Methods 0.000 claims abstract description 55
- 238000004364 calculation method Methods 0.000 claims abstract description 21
- 238000000034 method Methods 0.000 claims abstract description 14
- 238000004043 dyeing Methods 0.000 claims abstract description 6
- 238000004088 simulation Methods 0.000 claims abstract description 6
- 230000032823 cell division Effects 0.000 claims description 8
- 230000000694 effects Effects 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 claims description 2
- 238000011161 development Methods 0.000 abstract description 6
- 238000005192 partition Methods 0.000 abstract description 2
- 238000012545 processing Methods 0.000 abstract description 2
- 238000007447 staining method Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 19
- 230000005764 inhibitory process Effects 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 4
- 230000002900 effect on cell Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000011369 optimal treatment Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
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Abstract
The invention discloses a quick test method for biological effect of a medium-frequency micro-electric field, which arranges electrodes of a waveform generator at proper positions around a culture dish to generate a calculable uneven electric field in the culture dish; calculating the electric field distribution in the culture dish by using a finite element method; selecting test parameters according to the calculation result, and performing intermediate frequency micro-electric field biological effect test on the cells in the culture dish; processing the cells by adopting a staining method, and dividing an effective area and an ineffective area according to a color development result; and comparing the color development result with the electric field calculation result to obtain the effective electric field parameter range. The rapid test method has the advantages that the required device is simple, the principle is reliable, and the purpose of simultaneously carrying out the test of multiple groups of electric field parameters is realized by applying an uneven electric field in the test area; the test speed is greatly accelerated by a mode of dyeing partition and comparing with a simulation calculation result.
Description
Technical Field
The invention belongs to the technical field of biological effect tests, and particularly relates to a method for rapidly testing biological effect of a medium-frequency micro-electric field.
Background
It was found that the application of a medium frequency micro-electric field to tumor tissue destroys rapidly proliferating tumor cells without significant effect on normal cells. At present, related researches are still in a starting stage, and a large number of experiments are needed to obtain the biological effect of the medium-frequency micro-electric field acting on various tumor cells; in addition, in order to achieve a personalized treatment for the patient, it is necessary to quickly obtain the optimal treatment electric field parameters for a specific cell. Wherein the medium-frequency micro electric field is an electric field with the frequency of 10kHz-1MHz and the electric field intensity of 1V/cm-3V/cm.
The existing intermediate frequency micro-electric field biological effect test usually adopts a method of applying a uniform electric field to tumor cells, each test can only obtain the effect of a group of electric field parameters on the cells, and related test researchers need to perform a large amount of repeated operations to obtain the biological effect of different electric field parameters on a certain tumor cell or a certain tumor cell, so that the test efficiency is low, and the research period is long.
Disclosure of Invention
The invention provides a method for rapidly testing biological effects of an intermediate frequency micro-electric field, which can simultaneously test different electric field parameters of certain or a plurality of cells and rapidly obtain the biological effects of the intermediate frequency micro-electric field with different parameters on tumor cells.
In order to achieve the aim, the invention provides a rapid test method for biological effect of an intermediate frequency micro-electric field, which utilizes an electric field waveform generator to generate a calculable uneven electric field in a culture dish; calculating the electric field distribution in the culture dish; selecting test parameters according to the electric field distribution calculation result, and performing intermediate frequency micro electric field biological effect test on the cells in the culture dish; dividing an effective area and an ineffective area according to a test result; and comparing the test result with the electric field distribution calculation result to obtain the electric field parameter range with the inhibition effect.
Further, the method comprises the following steps:
and 5, comparing the area division result of the step 4 with the electric field calculation result of the second step to obtain the electric field parameter range of the effective area.
Further, in step 1, a plurality of culture dishes which are the same in size and are coaxially arranged are arranged.
Further, in step 4, the cells in the culture dish after the experiment are treated by staining to divide the effective area and the ineffective area.
Further, in step 2, finite element software is used for solving the electric field intensity range in the culture dish corresponding to the applied different voltages.
Compared with the prior art, the invention has at least the following beneficial technical effects:
the device required by the rapid test method is simple in structure, convenient to use and capable of being flexibly adjusted according to test requirements, the principle of calculating the electric field intensity by adopting a finite element method is reliable, the result is accurate, the universality is high, test environments with different electric field parameters are obtained by applying an uneven electric field to a test area, the purpose of simultaneously carrying out multiple groups of electric field parameter tests is achieved, the cell division condition in a culture dish after the test is analyzed by adopting a dyeing method, an area which has an inhibiting effect on cell division in the test culture dish is obtained, and compared with the calculation result, the effective electric field intensity range can be intuitively and accurately obtained. The method saves the cost of manpower, material resources and time, so that the test can be rapidly carried out, and the application value of the test result and the efficiency of related medical research can be greatly improved.
Further, in step 1, a plurality of culture dishes with the same size and coaxial arrangement are arranged, and different types of cells are cultured in different layers, so that the function of simultaneously performing different types of cell tests is realized.
Furthermore, the biological effect of the electric field with different intensities acting on the cells is obtained by comparing the dyeing subareas with the simulation calculation result, and the test speed is greatly accelerated on the premise of ensuring good test effect and reliable result.
Drawings
FIG. 1 is a schematic three-dimensional perspective view of an electrode placement location;
FIG. 2 is a spatial electric field distribution diagram inside the culture dish;
FIG. 3 is a schematic view of example 3.
In the drawings: 1. a first electrode, 2, a second electrode, 3, an electric field waveform generator, 4, a culture dish.
Detailed Description
In order to make the objects and technical solutions of the present invention clearer and easier to understand. The present invention will be described in further detail with reference to the following drawings and examples, wherein the specific examples are provided for illustrative purposes only and are not intended to limit the present invention.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention. Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified. In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Example 1
Referring to fig. 1, a method for rapidly testing biological effect of a medium-frequency micro-electric field comprises the following steps:
in the first step, as shown in fig. 1, the electrodes at the two output ends of the electric field waveform generator 3 are a first electrode 1 and a second electrode 2, the first electrode 1 is a cylindrical rod-shaped electrode, the second electrode 2 is a cylindrical surface, the first electrode 1 is arranged on the axis of the culture dish 4, the second electrode 2 is arranged outside the culture dish 4, and the first electrode 1, the second electrode 2 and the culture dish 4 are arranged coaxially.
And secondly, establishing an electric field simulation calculation model of the device shown in the figure 1 according to the sizes and the arrangement modes of the culture dish and the electrodes used in the first step, and solving by using finite element software to obtain the electric field intensity ranges in the culture dish 4 corresponding to the applied different voltages.
Thirdly, selecting the output waveform of the electric field waveform generator 3 according to the electric field frequency and the electric field intensity range concerned by the micro electric field biological effect test and based on the calculation result of the second step, and carrying out the intermediate frequency micro electric field biological effect test on the cells in the culture dish; for example, the experiment focused on the biological effect of the electric field strength in the interval of 1V/cm-4V/cm on the cells, and according to the calculation result of the second step, as shown in FIG. 2, when the applied voltage is 10V, the spatial electric field distribution range of the main area in the culture dish is 1V/cm-4V/cm, the electric field frequency is 100kHz, so that the output waveform of the electric field waveform generator 3 is adjusted to 10V, and the medium frequency micro electric field biological effect test is performed on the cells in the culture dish.
And fourthly, treating the cells in the culture dish after the test in the third step by adopting a dyeing method, and dividing an effective area and an ineffective area according to a color development result. The region with obvious inhibition effect on cell division is the effective region, and the specific characteristics need to be determined according to the material selected by the test and the related biological test method.
And fifthly, comparing the color development result of the fourth step with the electric field calculation result of the second step or the spatial electric field distribution range of the third step, and judging which electric field range is effective for cell division in the electric field distribution range of 1V/cm-4V/cm in the culture dish to obtain an effective electric field range, wherein the electric field parameters of the region with obvious inhibition effect on cell division can be used as electric field parameters for treating tumors. If the effective electric field strength needs to be further refined, the above experiment can be performed again within the effective electric field range obtained in the fifth step.
If there is no inhibition effect on cell division in the spatial electric field distribution range selected in step 3, the voltage and frequency applied in the second step are adjusted to test the inhibition effect of other electric field frequencies and/or electric field intensity ranges on cell division.
Example 2
This example is the same as example 1 except that: in this embodiment, a plurality of culture dishes 4 having the same size and being coaxially arranged are arranged, and different kinds of cells are placed in different culture dishes 4.
Example 3
Referring to fig. 3, the intermediate frequency micro electric field biological effect test device comprises a multi-layer culture dish structure, a central rod electrode 1, a cylindrical surface electrode 2 and an electric field waveform generation module 3, wherein a circular mounting hole with the same size as the cross section of the rod electrode 1 is formed in the center of the multi-layer culture dish structure, the central rod electrode 1 is arranged in the mounting hole in the center of the multi-layer culture dish structure, the cylindrical surface electrode 2 is coaxial with the center line of the culture dish structure, the cylindrical surface electrode 2 surrounds the multi-layer culture dish structure, and two output ends of the electric field waveform generation module 3 are respectively connected to the central rod electrode 1 and the cylindrical surface electrode 2 through leads.
The multi-layer culture dish structure comprises a plurality of layers of culture dishes 4 which are sequentially arranged along the vertical direction and have the same specification, and the number of layers of the culture dishes 4 can be selected according to the requirement. The culture dish 4 is used for placing culture solution and cells for biological tests.
The diameter of the culture dish is 3cm-10cm, and the cylindrical surface electrode is preferably tightly attached to the outer wall of the culture dish.
Preferably, the length of the central rod electrode 1 and the cylindrical surface electrode 2 is more than 10cm greater than the height of the multi-layer culture dish structure, and the upper end and the lower end of the central rod electrode and the cylindrical surface electrode respectively exceed the top surface and the bottom surface of the culture dish by more than 5 cm.
The invention discloses a quick test method for biological effect of a medium-frequency micro-electric field, which comprises a culture dish, a waveform generator and an arrangement mode of the waveform generator, and the test method comprises the following steps: disposing a waveform generator electrode at a suitable location about the culture dish to generate a calculable non-uniform electric field in the culture dish; calculating the electric field distribution in the culture dish by using a finite element method; selecting test parameters according to the calculation result, and performing intermediate frequency micro-electric field biological effect test on the cells in the culture dish; processing the cells by adopting a staining method, and dividing an effective area and an ineffective area according to a color development result; and comparing the color development result with the electric field calculation result to obtain the effective electric field parameter range. The rapid test method has the advantages that the required device is simple, the principle is reliable, and the purpose of simultaneously carrying out the test of multiple groups of electric field parameters is realized by applying an uneven electric field in the test area; the test speed is greatly accelerated by a mode of dyeing partition and comparing with a simulation calculation result.
What has been described above are merely some embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made without departing from the inventive concept thereof, and these changes and modifications can be made without departing from the spirit and scope of the invention.
Claims (5)
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115125143A (en) * | 2022-08-15 | 2022-09-30 | 南京大学 | A cell culture system with real-time adjustable field strength and frequency |
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| CN1234069A (en) * | 1997-06-10 | 1999-11-03 | 塞托·帕尔斯科技公司 | Method and apparatus for handling substances with variable direction electric fields |
| US20030113832A1 (en) * | 2001-12-14 | 2003-06-19 | Lauf Robert J. | Apparatus and method for assaying electrophysiological effects |
| CN1566951A (en) * | 2003-06-16 | 2005-01-19 | 中国科学院大连化学物理研究所 | Cell sorting method based on micro flow control chips and special-purpose chips therefor |
| US20080124778A1 (en) * | 2006-11-29 | 2008-05-29 | Bio-Rad Laboratories, Inc. | Electroporation Cuvette with High Field Intensity and Field-Induced Agitation |
| CN102488967A (en) * | 2004-12-27 | 2012-06-13 | 斯坦顿有限公司 | Treating a tumor or the like with electric fields at different orientations |
| US20160346536A1 (en) * | 2000-02-17 | 2016-12-01 | Novocure Limited | Treating Bacteria with Electric Fields |
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2020
- 2020-12-30 CN CN202011626679.7A patent/CN112834573A/en active Pending
Patent Citations (7)
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|---|---|---|---|---|
| CN1234069A (en) * | 1997-06-10 | 1999-11-03 | 塞托·帕尔斯科技公司 | Method and apparatus for handling substances with variable direction electric fields |
| US20160346536A1 (en) * | 2000-02-17 | 2016-12-01 | Novocure Limited | Treating Bacteria with Electric Fields |
| US20030113832A1 (en) * | 2001-12-14 | 2003-06-19 | Lauf Robert J. | Apparatus and method for assaying electrophysiological effects |
| CN1566951A (en) * | 2003-06-16 | 2005-01-19 | 中国科学院大连化学物理研究所 | Cell sorting method based on micro flow control chips and special-purpose chips therefor |
| CN102488967A (en) * | 2004-12-27 | 2012-06-13 | 斯坦顿有限公司 | Treating a tumor or the like with electric fields at different orientations |
| US20080124778A1 (en) * | 2006-11-29 | 2008-05-29 | Bio-Rad Laboratories, Inc. | Electroporation Cuvette with High Field Intensity and Field-Induced Agitation |
| CN110669664A (en) * | 2019-10-22 | 2020-01-10 | 南方电网科学研究院有限责任公司 | Electric field treatment test method and device for algae on surface of silicone rubber |
Non-Patent Citations (1)
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115125143A (en) * | 2022-08-15 | 2022-09-30 | 南京大学 | A cell culture system with real-time adjustable field strength and frequency |
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Application publication date: 20210525 |