KR20220107586A - Detection Device for Cell Population using Electric Signal and Detection Method using the same - Google Patents

Abstract

The present invention relates to a cell count detection device using an electric signal and, more specifically, to a cell count detection method capable of identifying the number of cells during culture in real-time using only an electric signal without a microscope and dyeing treatment. The cell count detection device using an electrical signal according to the present invention comprises: a culture medium accommodating part accommodating a culture medium which is a cell count detection target; a cell chip immersed in the culture medium of the culture medium accommodating part and adsorbing cells in the culture medium; a measurement part measuring the capacitance of the cells absorbed by the cell chip and the culture medium; and an analysis part analyzing the cell area and the cell count based on the measured capacitance value.

Description

Device for detecting the number of cells using an electric signal and a detection method using the same

The present invention relates to an apparatus for detecting the number of cells using an electrical signal, and more particularly, to a method for detecting the number of cells in real time by using only an electrical signal without a microscope and staining treatment for the number of cells in culture.

In general, cell-based tests using cells, which are the basic unit of the living body, are used in a variety of ways, such as cell culture regulators and process research, disease diagnosis and treatment such as tumors and viral infections, new drug candidate material development and toxic substances, harmful substances, and environmental pollution tests. being used for the purpose.

Checking the cell population change and proliferation characteristics in the cell culture process is an important task in establishing optimal cell culture conditions. It was complicated and it took a long time to detect, so it was not efficient. Also, it was impossible to check the number of cells in real time.

Domestic Registered Patent No. 10-2189517 Domestic Patent Publication No. 10-2018-0129927

An object of the present invention for solving the above problems is to provide a method for detecting the number of cells in real time that can be detected in real time only with an electrical signal without a microscope and staining treatment for the number of cells in culture.

A cell number detection device using an electrical signal of the present invention for solving the above problems includes: a culture medium receiving unit for accommodating a cell number detection target culture medium; and a cell chip supported in the culture medium of the culture medium receiving unit, to which cells in the culture medium are adsorbed; and It includes; a measuring unit for measuring the capacitance of the cells and the culture medium adsorbed to the cell chip; and an analysis unit for analyzing the area of cells and the number of cells based on the measured capacitance value.

The cell chip includes a transparent metal oxide layer deposited on a substrate and a metal layer laminated on the transparent metal oxide layer.

The method for detecting the number of cells using an electrical signal according to the present invention comprises: a culture medium receiving step of receiving a culture medium to be detected in a cell number in a culture medium receiving unit; and a cell chip supporting step of supporting a cell chip for adsorbing cells in the culture medium receiving unit; It includes a measuring step of measuring the capacitance of the cells and the culture medium adsorbed to the cell chip; and an analysis step of analyzing the area of cells and the number of cells based on the measured capacitance value.

The cell chip of the cell chip loading step includes a transparent metal oxide layer deposited on a substrate and a metal layer laminated on the transparent metal oxide layer.

As described above, according to the device for detecting the number of cells using an electrical signal according to the present invention, the number of cells during culture can be detected in real time only with an electrical signal without a microscope and staining process, so that changes in the cell population can be efficiently monitored. .

1 is a block diagram showing the configuration of a cell number detection device using an electrical signal according to the present invention.
2 is a cell chip configuration of the cell number detection device using an electrical signal according to the present invention.
3 is a flowchart showing a method for detecting the number of cells using an electrical signal according to the present invention.
Figure 4 is an embodiment using the cell number detection device of the present invention, the change in impedance before and after loading the cell chip in the culture medium.
5 is a cell area ratio derivation result of an embodiment using the cell number detection device of the present invention.

Specific features and advantages of the present invention will be described in detail below with reference to the accompanying drawings. Prior to this, if it is determined that the detailed description of the function and its configuration related to the present invention may unnecessarily obscure the gist of the present invention, the detailed description will be omitted.

1 is a block diagram showing the configuration of a cell number detection device using an electrical signal according to the present invention, FIG. 2 is a cell chip configuration of the cell number detection device using an electrical signal according to the present invention, and FIG. It is a flowchart showing a cell number detection method using an electrical signal according to It is the result of deriving the cell area ratio of the Example using the cell number detection device.

The present invention relates to an apparatus for detecting the number of cells using an electrical signal, and more particularly, to a method for detecting the number of cells in real time by using only an electrical signal without a microscope and staining treatment for the number of cells in culture.

1 is a block diagram showing the configuration of an apparatus for detecting the number of cells using an electrical signal according to the present invention.

The cell number detection device using the electrical signal of the present invention is supported in the culture solution receiving part 100 and the culture solution receiving part for accommodating the cell number detection target culture medium, and the cell chip 200 to which cells in the culture medium are adsorbed and the cells It includes a measuring unit 300 for measuring the capacitance of the cells and the culture medium adsorbed to the chip, and an analyzing unit 400 for analyzing the area of cells and the number of cells based on the measured capacitance value.

2 is a cross-sectional view showing a stacked structure of a cell chip of an apparatus for detecting the number of cells using an electrical signal according to the present invention, wherein the cell chip 200 includes a transparent metal oxide layer 220 deposited on a substrate 210 and the and a metal layer 230 laminated on the transparent metal oxide layer.

The substrate 210 may be a resin material of any one of PET, PC, PES, PEN, or a combination thereof, and the transparent metal oxide 120 may be indium tin oxide (ITO), zinc oxide (ZnO), or indium zinc. It may be selected from any one of oxide (IZO), aluminum zinc oxide (AZO), zinc tin oxide (ZTO), or a combination thereof. Indium tin oxide (ITO), which is easy to apply, is used.

The method of depositing the transparent metal oxide 220 on the substrate may use any one of a physical method (Physical Vapor Deposition: hereinafter PVD), a chemical method (Chemical Vapor Deposition: hereinafter CVD), or a combination thereof. And, preferably, any one of the vacuum deposition method, the sputter method, and the ion plating method can be used.

The metal layer 230 is formed by sputtering gold (Au). At this time, when a metal other than gold is used, an ohmic junction with ITO is not made due to a difference in conduction band, so a problem may occur in the sensor.

The metal layer 230 is protected from contact with the cell culture solution by the protective layer 240, and by forming the protective layer, the gold portion is exposed to the cell culture solution, thereby preventing the current from escaping and a short circuit. .

The protective layer 240 may be made of any one of silicon-based and epoxy-based materials, and the protective layer may be formed to cover even a wire connection portion connected to the metal layer of the metal layer.

In addition, the protective layer 240 is configured to cover the surface of the metal layer, it is preferable to increase the accuracy of the sensor by covering the ITO to a minimum.

The measurement unit 300 is connected to the cell chip to measure the capacitance of the cells and the culture medium adsorbed to the cell chip, and the analysis unit 400 analyzes the area of cells and the number of cells based on the measured capacitance value. do.

Hydrogen atoms are adsorbed to the surface of the transparent metal oxide layer of the cell chip supported in the culture medium, and as carriers are diffused from the metal layer, the impedance decreases.

On the other hand, the action magnitude of the capacitance is proportional to the dielectric constant, which is an intrinsic property of a pure material, and is derived from the dielectric constant and area of the material adsorbed to the cell chip.

At this time, the measuring unit measures the capacitance per surface area of the culture medium and the capacitance per surface area of the cells, and through this, the cell area can be derived.

Equation 1 below is a calculation formula for calculating the total capacitance.

[Equation 1]

Total capacitance = (capacitance per area of cells * cell area) + {capacitance per area of solution * (total area of transparent metal oxide - area of cells)}

Accordingly, it is possible to monitor the death of cells and the increase in the number of cells in real time through the detection device and the detection method.

3 is a flowchart showing a method for detecting the number of cells using an electrical signal according to the present invention.

The method for detecting the number of cells using an electrical signal according to the present invention uses the cell number detection device using the above-described electrical signal. The cell chip loading step (S200) of loading the cell chip for the cell chip into the culture medium receiving unit, the measuring step (S300) of measuring the capacitance of the cells and the culture medium adsorbed to the cell chip, and the area and cells of the cells based on the measured capacitance value It includes an analysis step (S400) of analyzing the number of individuals.

A detailed description thereof has been described above, and a description thereof will be omitted.

Hereinafter, the present invention will be described in detail below with reference to a preferred embodiment. However, the following examples are intended to specifically illustrate the present invention, and are not limited thereto.

1. Preparation of cell chips

PET as a substrate, ITO as a transparent metal oxide, and gold (Au) as a metal layer were sequentially laminated. ITO was deposited on the substrate PET using a stuttering method, and ITO and gold were bonded using PDMS (polydimethylsiloxane). .

2. Preparation of the culture medium

The culture medium was prepared by adding an electrolyte liquid, a medium, etc. together to create an environment for culturing cells.

3. Change in Impedance

4 shows the change in impedance before and after loading the cell chip in the culture medium, blue is the applied signal, red is the impedance change before loading the cell chip, and yellow is the impedance change after loading the cell chip.

As a result, it was shown that the impedance decreased rapidly after the cell chip was loaded in the culture medium, which was determined to be due to the rapid decrease in resistance as hydrogen atoms were adsorbed on the ITO surface and diffusion of carriers from gold occurred.

4. Reliability of cell area measurement

Thereafter, the cells to be cultured were added to the culture medium and cultured, and the area of the cells was checked after 2 to 3 hours after the cells were added.

In order to check whether the detection device detects or not, the cell area ratio was calculated after the actual cells were put into the culture medium in a weight ratio of 1:2 under the same conditions.

At this time, the capacitance was determined by signal detection through an oscilloscope and analysis through MATLAB. The graph was analyzed after observing the charge/discharge time with an oscilloscope by giving a 500 mv, 0.5 pulse width signal to both ends with a function generator. 5 shows the results of deriving the cell area ratio, and the ratio of area 1 to area 2 was measured to be about 1: 2, which was determined to have reliability.

As described above, the present invention has been mainly described with reference to the accompanying drawings, but those of ordinary skill in the art to which the present invention pertains within the scope not departing from the technical spirit and scope described in the claims of the present invention. Various modifications or variations of the present invention can be practiced. Accordingly, the scope of the present invention should be construed by the appended claims including examples of many such modifications.

100: culture medium receiving part
200: cell chip
210: description
220: transparent metal oxide layer
230: metal layer
240: protective layer
300: measurement unit
400: analysis unit

Claims (4)

A culture solution receiving unit for accommodating the cell number detection target culture solution; and
A cell chip supported in the culture solution of the culture solution receiving part, to which cells in the culture solution are adsorbed; and
A measuring unit for measuring the capacitance of the cells and the culture medium adsorbed to the cell chip; and
An analysis unit that analyzes the area of cells and the number of cells based on the measured capacitance value; characterized in that it comprises a
A device for detecting the number of cells using an electrical signal.
The method of claim 1,
The cell chip is
a transparent metal oxide layer deposited on the substrate; and
and a metal layer laminated on the transparent metal oxide layer.
A device for detecting the number of cells using an electrical signal.
A culture medium receiving step of receiving the culture medium to be detected in the cell number in the culture medium receiving unit; and
A cell chip loading step of loading a cell chip for adsorbing cells to the culture medium receiving unit; and
A measuring step of measuring the capacitance of the cells and the culture medium adsorbed to the cell chip; and
An analysis step of analyzing the area of cells and the number of cells based on the measured capacitance value; characterized in that it comprises
A method for detecting the number of cells using electrical signals.
The method of claim 1,
The cell chip of the cell chip loading step is
a transparent metal oxide layer deposited on the substrate; and
and a metal layer laminated on the transparent metal oxide layer.
A method for detecting the number of cells using electrical signals.

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