JP2021179319A - Analysis method and analyzer - Google Patents

Abstract

To provide a technology that can effectively analyze and evaluate time-series data of measurement signals of a biological specimen using a small number of parameters.SOLUTION: An analysis method includes: a step for analyzing a power spectrum of a measurement signal and for acquiring a low-frequency spectrum that is a power spectrum distribution in a low frequency region; a step for acquiring parameters from the low-frequency spectrum; and a step for evaluating the measurement signal based on the parameters. The measurement signal is a continuous signal for at least one minute or more. The low frequency region is included in the range of 2 Hz or less. The parameters include: a peak frequency that is a frequency at which a power spectral density is maximum; and a peak power that is the power spectral density at the peak frequency. In this way, a long-term change can be simply and easily analyzed and evaluated by focusing on a few parameters.SELECTED DRAWING: Figure 5

Description

The present invention relates to an analysis method and an analysis device for a measurement signal in a compound response to a sample of biological origin.

Conventionally, in order to evaluate a compound response to a biological sample, measurement such as electrical measurement or image measurement is performed, and time-series data of the obtained measurement signal is analyzed.

As an example, in order to measure the induced electricity of cells or tissues such as nerve cells, a MEA electrode, which is a cell holding container having a plurality of electrodes provided on the bottom surface, called a micro-electrode array (MEA), is used. A system for measuring extracellular potential using it is known. The MEA electrode is described in, for example, Patent Document 1. In such a MEA electrode, a cell layer or a section of tissue is arranged on the electrode, and the extracellular potential of the cells contained therein is measured.

As another example, measurement of time-series signals such as electroencephalograms and electrocardiograms is also well known. In vitro and in vivo, using various bioimaging methods such as fluorescence imaging including calcium imaging and nuclear magnetic resonance imaging (MRI) and nuclear medicine imaging (PET, etc.) used as clinical methods. Measurement is performed using time-series image data.

Japanese Patent Publication No. 2002-523726

In the analysis of such various time-series data, if the evaluation index is not established, a number of analysis parameters are calculated, which parameter is effective as the evaluation index, or how to combine multiple parameters. If so, it is necessary to find out whether it can be an evaluation index.

For example, when analyzing the extracellular potential obtained by the MEA measurement method for comparison of compound responses, more than 50 parameters are calculated from the time-series signal of the extracellular potential, and a heat map is created for each parameter for each compound. Then, a method of making a comparison is known. In such a method, it is necessary to calculate a large number of parameters, and there is a problem that it takes a lot of time and effort for analysis.

In particular, the short-term response in the compound response is easily visualized in graphs and images and is easily noticed as a parameter, whereas the long-term response is difficult to be focused on.

The present invention has been made in view of such circumstances, and an object of the present invention is to provide a technique for effectively analyzing and evaluating time-series data of measurement signals of biological samples using a small number of parameters.

In order to solve the above problems, the first invention of the present application is a method for analyzing a measurement signal in a compound response to a sample of biological origin. A) Power spectrum analysis of the measurement signal is performed, and power spectrum distribution in a low frequency region is performed. It has a step of acquiring a low frequency spectrum, b) a step of acquiring a parameter from the low frequency spectrum, and c) a step of evaluating the measurement signal based on the parameter. The signal is a continuous signal for at least 1 minute or more, the low frequency region is included in the range of 2 Hz or less, and the parameters are the peak frequency, which is the frequency at which the power spectral density is maximum, and the power at the peak frequency. Includes peak power, which is the spectral density.

The second invention of the present application is the analysis method of the first invention, and the parameter further includes a peak potential which is a square root of the peak power.

The third invention of the present application is a method for analyzing a measurement signal in a compound response to a biological sample. A) The power spectrum of the measurement signal is analyzed to obtain a low frequency spectrum which is a power spectrum distribution in a low frequency region. The step of acquiring the parameter from the low frequency spectrum, and c) the step of evaluating the measurement signal based on the parameter, and the measurement signal is at least 1 minute or longer. The low frequency region is included in the range of 2 Hz or less, and the parameter includes area power which is the total power spectral density in the low frequency region.

The fourth invention of the present application is the analysis method according to any one of the first invention to the third invention, and the low frequency region is a region of 1 Hz or less.

The fifth invention of the present application is the analysis method according to any one of the first invention to the fourth invention, and the measurement signal is a continuous signal for at least 10 minutes or more.

The sixth invention of the present application is the analysis method of any one of the first invention to the fifth invention, and the step c) is c1) a step of performing clustering using a plurality of the parameters, and c2) for each of the clusters. Including the process of performing the evaluation of.

The seventh invention of the present application is the analysis method according to any one of the first invention to the sixth invention, and the measurement signal is an electric signal of a cell group in which nerve cells and glial cells are co-cultured.

The eighth invention of the present application is the analysis method of the seventh invention, and in the step c), the peak power is used as the parameter, and the neuroconvulsant toxicity is evaluated.

The ninth invention of the present application is the analysis method according to any one of the first invention to the sixth invention, and the measurement signal is a signal relating to fluorescence intensity in calcium imaging of cells using a fluorescent dye.

The tenth invention of the present application is a device for analyzing a measurement signal in a compound response to a biological sample, and analyzes the power spectrum of the measurement signal to obtain a low frequency spectrum which is a power spectrum distribution in a low frequency region. The measurement signal has a frequency spectrum acquisition unit, a parameter acquisition unit that acquires parameters from the low frequency spectrum, and an analysis unit that analyzes and / or evaluates the measurement signal based on the parameters. , The low frequency region is included in the range of 2 Hz or less, and the parameters are the peak frequency, which is the frequency at which the power spectral density is maximum, and the power spectral density at the peak frequency. Including peak power, which is.

The eleventh invention of the present application is the analysis apparatus of the tenth invention, and the parameter further includes a peak potential which is a square root of the peak power.

The twelfth invention of the present application is a device for analyzing a measurement signal in a compound response to a biological sample, and analyzes the power spectrum of the measurement signal to obtain a low frequency spectrum which is a power spectrum distribution in a low frequency region. The measurement signal has a frequency spectrum acquisition unit, a parameter acquisition unit that acquires parameters from the low frequency spectrum, and an analysis unit that analyzes and / or evaluates the measurement signal based on the parameters. , Which is a continuous signal of at least 1 minute or longer, the low frequency region is included in the range of 2 Hz or less, and the parameter includes area power which is the total power spectral density in the low frequency region.

According to the first to twelfth inventions of the present application, it is possible to easily analyze and evaluate long-term changes by paying attention to a small number of parameters regarding the low frequency spectrum of the measurement signal.

It is a perspective view of the container with MEA electrode which concerns on 1st Embodiment. It is a schematic diagram of the analysis system which concerns on 1st Embodiment. It is a figure which showed an example of the original waveform of the field potential of a co-cultured nerve cell. It is a figure which showed an example of the correction waveform of the field potential of a co-cultured nerve cell. It is a flowchart which shows the flow of the analysis process of the field potential of the co-cultured nerve cell which concerns on 1st Embodiment. It is a figure which showed an example of a low frequency spectrum. It is a figure which showed the peak potential for each added compound of the co-cultured nerve cell obtained by an experiment. It is a figure which showed the peak potential for each added compound of the co-cultured nerve cell obtained by an experiment. It is a figure which showed the peak potential for each added compound of the co-cultured nerve cell obtained by an experiment. It is a flowchart which shows the flow of the analysis processing of the measurement signal in the compound response to the sample of a biological origin which concerns on one modification. It is a flowchart which shows the flow of the analysis processing of the measurement signal in the compound response to the sample of a biological origin which concerns on other modification.

Hereinafter, embodiments of the present invention will be described with reference to the drawings. It should be noted that the drawings are shown schematically except for experimental data, and for convenience of explanation, some configurations in the drawings may be exaggerated. That is, the interrelationship between the sizes and positions of the configurations shown in the drawings is not always accurately described and can be changed as appropriate.

<1. First Embodiment>
<1-1. Overview of extracellular potential measurement>
The method for measuring the extracellular potential according to the first embodiment of the present invention will be described with reference to FIG. FIG. 1 is a perspective view of a container 1 with a MEA electrode used for measuring extracellular potential. The container 1 is a container for internally accommodating and retaining a cultured cell layer or sliced tissue and a liquid such as a medium or a storage solution, and measuring the electrical characteristics of the retained cell layer.

As shown in FIG. 1, the container 1 has a cup-shaped recess 10 in which cells and the like are housed. The container 1 has a plurality of electrodes on the bottom surface 11 of the recess 10 for measuring the electrical characteristics of the cells or tissues contained in the container 1. That is, the bottom surface 11 is the measurement surface.

The container 1 of the present embodiment is a bottomed cylindrical container having only one recess 10, but the present invention is not limited to this. MEA measurement may be performed using a so-called multi-well plate having a plurality of recesses.

A plurality of working electrodes 21 and a plurality of reference electrodes 22 are arranged on the bottom surface 11.

The plurality of working electrodes 21 are arranged near the center of the bottom surface 11. When measuring the extracellular potential, the cell layer or tissue to be measured is arranged on the working electrode 21. The plurality of working electrodes 21 are two-dimensionally arranged at intervals from each other in a plan view. In the present embodiment, 16 working electrodes 21 are arranged in a matrix of 4 rows and 4 columns, but the number and arrangement of the working electrodes 21 are not limited to this. Further, although the working electrode 21 of the present embodiment is square in the top view, the shape of the working electrode 200 may be circular, rectangular, or any other shape.

Each of the reference electrodes 22 is arranged on the edge side of the bottom surface 11 with respect to the working electrode 21. When measuring the extracellular potential, the reference electrode 22 is arranged so that the cell layer or tissue to be measured does not come into contact with the reference electrode 22. Although the number of reference electrodes 22 in this embodiment is two, it may be one or three or more. Further, although the shape of the reference electrode 22 of the present embodiment is an arc shape, the shape of the reference electrode 22 may be a circle, a square, a rectangle, or any other shape in the top view.

When such a container 1 is used, for example, an electrical stimulus is applied between the working electrode 21 and the reference electrode 22 to the cell layer or tissue held in the container 1, and the response to the stimulus is observed. Can be done. Further, the compound response can be observed by adding various compounds to the cell layer or tissue held in the container 1 and observing the change in the extracellular potential (field potential) thereof.

<1-2. Measurement signal analysis system>
Next, the measurement signal analysis system 100 according to the present embodiment will be described with reference to FIG. As shown in FIG. 2, the analysis system 100 includes a container 1, a measuring device 3, an analysis device 4, and an operation unit 5.

The measuring device 3 is a device connected to the container 1 and for measuring the extracellular potential of the cell layer or tissue held in the container 1. As the measuring device 3, for example, a dedicated device used for the MEA measuring method is used. The measuring device 3 has a measuring signal acquisition unit 31 connected to the working electrode 21 and the reference electrode 22 of the container 1. The measurement signal acquisition unit 31 acquires the field potential of the cell layer or tissue as a measurement signal from the potential difference between the working electrode 21 and the reference electrode 22.

The analysis device 4 is a device for analyzing the measurement signal measured by the measurement device 3. The analysis device 4 is realized by, for example, a personal computer. The analysis device 4 is composed of an arithmetic processing unit such as a CPU, a memory such as a RAM, and a storage unit such as a hard disk drive.

The analysis device 4 reads the computer program or data stored in the storage unit into the memory, and the arithmetic processing unit performs arithmetic processing based on the computer program or data, whereby the analysis processing described later can be performed.

Further, the analysis device 4 is connected to an operation unit 5 having a display unit 51 composed of a display device or the like and an input unit 52 composed of a keyboard, a mouse or the like. The analysis device 4 may be integrated with the measurement device 3.

The analysis device 4 has a low frequency spectrum acquisition unit 41, a parameter acquisition unit 42, and an analysis unit 43 as processing units realized on software.

The low frequency spectrum acquisition unit 41 analyzes the power spectrum of the measurement signal measured by the measuring device 3 and acquires the low frequency spectrum which is the power spectrum distribution in the low frequency region.

In the present embodiment, the low frequency spectrum acquisition unit 41 acquires the low frequency spectrum by extracting the low frequency region from the obtained power spectrum distribution after analyzing the power spectrum of the measurement signal. Not limited to. For example, the low frequency spectrum acquisition unit 41 may process the measurement signal with a low-pass filter and then perform power spectrum analysis of the processed measurement signal.

The parameter acquisition unit 42 acquires one or a plurality of parameters from the low frequency spectrum acquired by the low frequency spectrum acquisition unit 41. The parameter acquisition unit 42 of the present embodiment acquires at least three parameters of peak frequency, peak power, and peak potential.

Here, the peak frequency is the frequency at which the power spectral density is maximum in the low frequency spectrum. Peak power is the power spectral density at the peak frequency. The peak potential is the square root of the peak power.

The analysis unit 43 analyzes and / or evaluates the measurement signal based on at least one of the parameters acquired by the parameter acquisition unit 42. The analysis unit 43 of the present embodiment displays parameters on the display unit 51 of the operation unit 5, and the user operates from the input unit 52 to assist the analysis and evaluation by the user. However, the analysis unit 43 may perform various analyzes using a plurality of parameters, and the analysis unit 43 may automatically perform the analysis without any user operation.

<1-3. Analysis method for compound response of cultured cells>
Subsequently, a method for analyzing the compound response of the cultured cells will be described with reference to FIGS. 3 to 6. In the present embodiment, as the cultured cells, those obtained by co-culturing nerve cells and glial cells (hereinafter referred to as "co-cultured nerve cells") are used. Further, as the compound, a plurality of compounds related to convulsive toxicity are used.

FIG. 3 is a diagram showing an example of the original waveform of the field potential of co-cultured nerve cells. FIG. 4 is a diagram showing an example of a corrected waveform of the field potential of co-cultured nerve cells. 3 and 4 show field potential waveforms for 5 seconds in each stage for four periods, respectively.

As shown in Fig. 3, the field potential of co-cultured neurons is a region where low-frequency swells and fluctuations in electrical signals called spike signals are large (in FIGS. 3 and 4) in the original waveform without manipulation. , With the reference numeral "S").

Conventionally, when analyzing the field potential of co-cultured nerve cells, in order to analyze the action potential of nerve cells, we focus on the region where the electrical signal becomes large, which is called the spike signal. Therefore, in a normal measuring device, as shown in FIG. 4, a corrected waveform corrected for low-frequency swell is displayed and output. Then, by analyzing the corrected waveform, the compound response and the like are analyzed and evaluated.

In the analysis method of the present embodiment, the field potential is analyzed by focusing on the low-frequency swell component that has been truncated by the conventional correction. That is, in the measuring device 3 of the present embodiment, the uncorrected field potential is acquired as a measurement signal without the correction for removing the low-frequency swell component. As a result, as described below, parameters valid for analysis can be obtained.

FIG. 5 is a flowchart showing the flow of the field potential analysis process of the co-cultured nerve cells in the present embodiment. The analysis process of the present embodiment is performed to add a plurality of types of agents to co-cultured nerve cells cultured under the same conditions at a plurality of concentrations and evaluate their nerve spasm toxicity.

When analyzing the field potential of co-cultured nerve cells, first, the measuring device 3 measures the field potential of the co-cultured nerve cells and acquires a measurement signal which is a continuous signal for at least 1 minute (step S110). The measurement signal of this embodiment is a continuous signal for at least 10 minutes or more. The sampling frequency of the measurement signal of this embodiment is 20 kHz.

Next, the measurement signal obtained by the measuring device 3 is input to the analysis device 4. Then, the low frequency spectrum acquisition unit 41 of the analysis device 4 analyzes the power spectrum of the measurement signal and acquires the power spectrum distribution (step S121).

Subsequently, the low frequency spectrum acquisition unit 41 acquires a low frequency spectrum obtained by extracting the power spectrum distribution in the low frequency region from the obtained power spectrum distribution (step S122).

Here, in the present application, the frequency band in which the maximum frequency is 2 Hz or less is referred to as a “low frequency region”. In particular, in the present embodiment, the low frequency region indicates a frequency band in which the maximum frequency is 1 Hz.

As described above, the low frequency spectrum acquisition step S120 for acquiring the low frequency spectrum, which is the power spectrum distribution of the measurement signal in the low frequency region, is configured by the steps S121 and S122 performed by the low frequency spectrum acquisition unit 41. As described above, the step S120 for acquiring the low frequency spectrum may be configured by a step different from the steps S121 and S122.

Subsequently, the parameter acquisition unit 42 detects a parameter (hereinafter referred to as “first parameter”) that can be directly derived from the low frequency spectrum (step S131). Specifically, in the present embodiment, as the first parameter in step S131, the peak frequency, which is the frequency at which the power spectral density in the low frequency region is maximized, and the peak power, which is the power spectral density in the peak frequency, are detected. .. In step S131, as the first parameter, a specific frequency which is a frequency having a power spectral density exceeding a predetermined threshold value (for example, 10% of the peak power) and an area power which is a total value of the power spectral densities in a low frequency region. However, they may be detected at the same time.

FIG. 6 is a diagram showing an example of a low frequency spectrum. The upper part of FIG. 6 is an example of a low frequency spectrum in the case where no drug is added to the co-cultured nerve cells. The lower part of FIG. 6 is an example of a low frequency spectrum when 30 μM of Gabazine, which is a kind of convulsive substance, is added to co-cultured nerve cells. FIG. 6 shows a peak frequency P and a peak power Q for each of the two low frequency spectra.

Then, the parameter acquisition unit 42 calculates a new parameter (hereinafter referred to as “second parameter”) based on the first parameter obtained in step S131 (step S132). Specifically, in the present embodiment, in step S132, the peak potential, which is the square root of the peak power, is calculated as the second parameter. In step S132, the dispersion degree and periodicity of a specific frequency may be calculated as the second parameter, or the area potential which is the square root of the area power or the value obtained by dividing the peak potential by the area potential may be calculated. good.

As described above, the parameter acquisition step S130 for acquiring the parameters for analyzing and evaluating the low frequency spectrum is configured by the steps S131 and S132 performed by the parameter acquisition unit 42. When the analysis and evaluation are performed using only the first parameter, the parameter acquisition step S130 may not include the step of acquiring the second parameter as in step S132.

Finally, the analysis unit 43 displays each parameter on the display unit 51, and analyzes and evaluates the measurement signal according to the command from the user input from the input unit 52 (step S140). In step S140, the measurement signal is analyzed and evaluated based on at least one of the parameters acquired in step S130.

<1-4. Effectiveness of the analysis method of this embodiment>
Using the above analysis method, in the present embodiment, the presence or absence of nerve spasm toxicity and the strength of the compound response of the co-cultured nerve cell in which the nerve cell and the glial cell are co-cultured are evaluated based on the peak potential. Will be done.

7 to 9 are diagrams showing the peak potentials of the added compounds of the co-cultured nerve cells obtained by the experiment, respectively. 7 to 9 show a bar graph for each administered drug. In each graph, the horizontal axis is an item, and the additive-free (control, Ctr) and the added compound concentration (for example, 0.1 μM, etc.) are described. The vertical axis shows the peak potential value.

FIG. 7 shows the case where picrotoxin, Gabazine, pentylenetetrazole (PTZ, Pentylenetetrazole), and acetaminophen were added to the dispersed culture of cerebral nerve cells extracted from rat fetuses. It shows the peak potential.

FIG. 8 shows picrotoxin, Gabazine, and 4-aminopyridine for a dispersed culture of thawed cells obtained by cryopreserving cerebral nerve cells extracted from commercially available rat fetuses. ), And the peak potential when acetaminophen is added.

FIG. 9 shows picrotoxin (Picrotoxin), 4-aminopyridine (4-aminopyridine), pentylenetetrazol (PTZ, Pentylenetetrazole), and a dispersion culture of commercially available human iPS-derived neurons (XCell Science). The peak potential when acetaminophen is added is shown.

According to the experimental results shown in FIG. 7, in picrotoxin, Gabazine and Pentylenetetrazole, which have neuroconvulsant toxicity, the peak potential tends to increase for each 10-fold increase in the concentration of the added compound. It was seen. On the other hand, in acetaminophen, which does not have neuroconvulsant toxicity, the peak potential does not tend to increase even if the concentration of the added compound increases.

In the experimental results shown in FIG. 8, in picrotoxin, Gabazine, and 4-aminopyridine, which have neuroconvulsant toxicity, the peak potential increases with each 10-fold increase in the concentration of the added compound. There was a tendency to do. On the other hand, in acetaminophen, which does not have neuroconvulsant toxicity, the peak potential tends to increase slightly as the concentration of the added compound increases, but no significant increase tendency is observed.

In the experimental results of FIG. 9, in picrotoxin having neuroconvulsant toxicity, the peak potential increased with the increase of the added compound concentration up to 1 μM, and then the change of the peak potential due to the increase of the added compound concentration was observed. I couldn't. In 4-aminopyridine, which has neuroconvulsant toxicity, the peak potential tended to increase as the concentration of the added compound increased. In Pentylenetetrazole, which has neuroconvulsant toxicity, the peak potential increased with the increase in the concentration of the added compound up to 0.3 mM, and then the peak potential changed with the increase in the concentration of the added compound. There wasn't. On the other hand, in acetaminophen, which does not have neuroconvulsant toxicity, the peak potential tends to increase slightly as the concentration of the added compound increases, but no significant increase tendency is observed.

From the above experimental results, regarding the compound response of co-cultured nerve cells in which nerve cells and glial cells are co-cultured, there is a certain correlation between the presence or absence and strength of nerve spasm toxicity and the magnitude relationship of the peak potential value. Is considered to have.

Therefore, in the field potential analysis process of the co-cultured nerve cells shown in FIG. 5 above, it is considered possible to evaluate the measurement signal in step S140 based on one parameter called peak potential. That is, by performing such an analysis process, time-series data of measurement signals of biological samples can be effectively analyzed and evaluated with a small number of parameters.

In particular, recent studies suggest that astrocytes, a type of glial cell, are significantly involved in neuroconvulsive seizures such as epilepsy. Waveform changes in field potential due to astrocyte-derived calcium dynamics are likely to affect the low-frequency region in particular. Therefore, it is considered that the parameters obtained from the low frequency spectrum of the field potential as shown in the above experimental results have a correlation with the presence / absence and strength of nerve spasm toxicity.

Not only the toxicity of nerve spasm, but also the measurement signals of samples derived from other organisms have been analyzed by focusing on visually characteristic signals such as spike signals. However, in the conventional method, the change of the measurement signal appearing in a long-term cycle has not been analyzed. Therefore, as in the present invention, by paying attention to the low frequency spectrum of the measurement signal and performing the evaluation using the parameters obtained from the low frequency spectrum, it is possible to easily analyze and evaluate the long-term change. In this way, it leads to effective analysis and evaluation with a small number of parameters from a viewpoint different from the conventional one.

<2. Modification example>
Although one embodiment of the present invention has been described above, the present invention is not limited to the above embodiment.

FIG. 10 is a flowchart showing the flow of analysis processing of the measurement signal in the compound response to the biological sample according to the modified example. In the example of FIG. 10, a low frequency spectrum acquisition including a step of acquiring a measurement signal (step S210), a step of performing power spectrum analysis (step S221), and a step of extracting a low frequency region of the power spectrum distribution (step S222). The step S220 is performed in the same manner as the step S110 of the first embodiment and the step S120 including the steps S121 and S122.

Subsequently, the parameter acquisition unit 42 detects the area power, which is a parameter that can be directly derived from the low frequency spectrum (step S230). Area power is the total value of the power spectral density in the low frequency region.

Then, the analysis unit 43 displays the area power, which is a parameter for analysis, on the display unit 51, and analyzes and evaluates the measurement signal according to the command from the user input from the input unit 52 (step S240). As in the example of FIG. 10, the parameter used for the analysis may be only the area power.

FIG. 11 is a flowchart showing the flow of analysis processing of the measurement signal in the compound response to the biological sample according to the other modification. In the example of FIG. 11, a low frequency spectrum acquisition including a step of acquiring a measurement signal (step S310), a step of performing power spectrum analysis (step S321), and a step of extracting a low frequency region of the power spectrum distribution (step S322). The step S320 is performed in the same manner as the step S110 of the first embodiment and the step S120 including the steps S121 and S122.

Subsequently, the parameter acquisition unit 42 detects a first parameter that can be directly derived from the low frequency spectrum (step S331). In the example of FIG. 11, as the first parameter, the sum of the peak frequency, which is the frequency at which the power spectral density is maximum in the low frequency region, the peak power, which is the power spectral density in the peak frequency, and the power spectral density in the low frequency region. Detects the area power, which is a value.

Then, the parameter acquisition unit 42 calculates the second parameter based on the first parameter obtained in step S331 (step S332). In the example of FIG. 11, as the second parameter, the peak potential which is the square root of the peak power and the area potential which is the square root of the area power are calculated.

After that, the analysis unit 43 performs clustering for a plurality of measurement signals using three types of parameters, peak frequency, peak potential, and area potential (step S341). At this time, the plurality of measurement signals include, for example, those having different experimental conditions such as the type of biological sample to be measured, the type and concentration of the added compound, and the like. By performing clustering, the relationship between the experimental conditions and the parameters appears.

Finally, the analysis unit 43 displays each parameter and the result of clustering on the display unit 51, and evaluates the measurement signal according to the command from the user input from the input unit 52 (step S342). As described above, the step of performing analysis and evaluation (step S340) may be composed of step S341 in which the analysis unit 43 automatically performs analysis and step S342 in which the user manually evaluates.

As in the example of FIG. 11, when the field potential of a co-cultured nerve cell in which a nerve cell and a glial cell are co-cultured is analyzed by performing clustering using a plurality of parameters, for example, the compound response Compounds with similar tendencies can be found. That is, it becomes possible to discover compounds having a similar mechanism of action that causes convulsions by the compounds by clustering. Using this method, it is possible to predict the mechanism of action of a compound whose mechanism of action is unknown.

As described above, the method for analyzing the measurement signal in the compound response to the biological sample of the present invention is a simple and easy-to-understand evaluation method using a single parameter as in the first embodiment and the example of FIG. It can also be used, and as in the example of FIG. 11, it can also be used as an evaluation method for the mechanism of action of the compound response.

Further, in the above embodiment, the analysis target is the field potential of co-cultured nerve cells in which nerve cells and glial cells are co-cultured, but the present invention is not limited to this. The analysis target of the present invention may be other types of measurement signals as long as they are measurement signals in the compound response to a sample of biological origin. The measurement signal to be analyzed may be, for example, an electric signal such as an electroencephalogram or an electrocardiogram.

Further, the measurement signal to be analyzed may be time-series data obtained from time-series image data obtained by using various bioimaging methods. An example of a measurement signal using the bioimaging method is a signal relating to the fluorescence intensity in calcium imaging of cells using a fluorescent dye. Specifically, for example, in the time-series image data of calcium imaging, the total value of the luminance in a specific region is analyzed as a time-series measurement signal.

Further, the elements appearing in the above-described embodiments and modifications may be appropriately combined as long as there is no contradiction.

4 Analytical unit 41 Low frequency spectrum acquisition unit 42 Parameter acquisition unit 43 Analysis unit

Claims (12)

It is a method of analyzing the measurement signal in the compound response to the sample of biological origin.
a) A step of performing power spectrum analysis of the measurement signal and acquiring a low frequency spectrum which is a power spectrum distribution in a low frequency region.
b) The process of acquiring parameters from the low frequency spectrum and
c) A step of evaluating the measurement signal based on the parameters and
Have,
The measurement signal is a continuous signal for at least 1 minute or longer.
The low frequency region is included in the range of 2 Hz or less, and is included in the range of 2 Hz or less.
The above parameters are
The peak frequency, which is the frequency at which the power spectral density is maximum, and
Peak power, which is the power spectral density at the peak frequency,
Analysis method including. The analysis method according to claim 1.
The above parameters are
An analysis method further comprising a peak potential which is the square root of the peak power. It is a method of analyzing the measurement signal in the compound response to the sample of biological origin.
a) A step of performing power spectrum analysis of the measurement signal and acquiring a low frequency spectrum which is a power spectrum distribution in a low frequency region.
b) The process of acquiring parameters from the low frequency spectrum and
c) A step of evaluating the measurement signal based on the parameters and
Have,
The measurement signal is a continuous signal for at least 1 minute or longer.
The low frequency region is included in the range of 2 Hz or less, and is included in the range of 2 Hz or less.
The above parameters are
An analysis method comprising area power, which is the sum of the power spectral densities in the low frequency region. The analysis method according to any one of claims 1 to 3.
The analysis method, wherein the low frequency region is a region of 1 Hz or less. The analysis method according to any one of claims 1 to 4.
The analysis method, wherein the measurement signal is a continuous signal for at least 10 minutes or longer. The analysis method according to any one of claims 1 to 5.
The step c) is
c1) A step of performing clustering using the plurality of parameters, and
c2) The process of evaluating each cluster and
Analysis method including. The analysis method according to any one of claims 1 to 6.
The analysis method, wherein the measurement signal is an electrical signal of a cell group in which nerve cells and glial cells are co-cultured. The analysis method according to claim 7.
An analysis method for evaluating neuroconvulsant toxicity using the peak power as the parameter in the step c). The analysis method according to any one of claims 1 to 6.
The analysis method, wherein the measurement signal is a signal relating to the fluorescence intensity in calcium imaging of cells using a fluorescent dye. An analyzer for measuring signals in compound responses to biological samples.
A low-frequency spectrum acquisition unit that analyzes the power spectrum of the measurement signal and acquires the low-frequency spectrum, which is the power spectrum distribution in the low-frequency region.
A parameter acquisition unit that acquires parameters from the low frequency spectrum,
An analysis unit that analyzes and / or evaluates the measurement signal based on the parameters, and
Have,
The measurement signal is a continuous signal for at least 1 minute or longer.
The low frequency region is included in the range of 2 Hz or less, and is included in the range of 2 Hz or less.
The above parameters are
The peak frequency, which is the frequency at which the power spectral density is maximum, and
Peak power, which is the power spectral density at the peak frequency,
Including analysis equipment. The analysis device according to claim 10.
The above parameters are
An analyzer further comprising a peak potential which is the square root of the peak power. An analyzer for measuring signals in compound responses to biological samples.
A low-frequency spectrum acquisition unit that analyzes the power spectrum of the measurement signal and acquires the low-frequency spectrum, which is the power spectrum distribution in the low-frequency region.
A parameter acquisition unit that acquires parameters from the low frequency spectrum,
An analysis unit that analyzes and / or evaluates the measurement signal based on the parameters, and
Have,
The measurement signal is a continuous signal for at least 1 minute or longer.
The low frequency region is included in the range of 2 Hz or less, and is included in the range of 2 Hz or less.
The above parameters are
An analyzer comprising area power, which is the sum of the power spectral densities in the low frequency region.

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