# US 12,595,456 B2 · Cell observation device, electrostimulation device, and cell observation method

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- Number: US 12,595,456 B2
- Document ID: US12595456B2
- Status: Granted
- Kind: B2
- Application number: 17/565,929
- Filed: 2021-12-30
- Published: 2022-04-21
- Granted: 2026-04-07
- Earliest claimed: 2014-01-16
- Assignees: Hamamatsu Photonics K.K.
- Applicants: Hamamatsu Photonics K.K.
- Inventors: Natsumi Saito; Taira Ito; Takuji Kataoka
- Web page: https://patentgrep.com/patent/US12595456B2

## CPC

- C12M 35/02: {Electrical or electromagnetic means, e.g. for electroporation or for cell fusion}
- C12M 41/36: {of biomass, e.g. colony counters or by turbidity measurements (electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404)}
- C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
- G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates}
- G01N 27/416: Systems (G01N27/27 takes precedence)
- G01N 33/4836: {using multielectrode arrays}
- G01N 33/48707: {by electrical means (G01N33/49, G01N33/493 take precedence)}
- G01N 33/48728: {Investigating individual cells, e.g. by patch clamp, voltage clamp (investigating individual particles in general G01N15/10)}
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A cell observation device is cell observation device for observing a cell held by a microplate having a well holding a sample including the cell and includes a microplate holder for holding the microplate thereon, an electrical stimulation unit including an electrode pair including a first electrode and a second electrode, and a position controller for controlling a position of the electrical stimulation unit in a state in which the first electrode is disposed closer to the center of the well than the second electrode when the electrode pair is disposed in the well of the microplate. The tip of the first electrode extends more than the tip of the second electrode.

## Claims

### Claim 1

A system for observing a cell comprising:
a sample case including at least one holder;
a dark box configured to accommodate the sample case including the at least one holder holding a sample including the cell, wherein the sample case having a bottom portion that is formed of a material which can transmit light from the sample;
a light-guiding optical system including a lens, the light-guiding optical system disposed under the bottom portion of the sample case and configured to guide a two-dimensional image;
a camera configured to capture the two-dimensional image that is guided by the light-guiding optical system and output movie data of the two-dimensional image;
an electrical stimulator including an electrode pair having a first electrode and a second electrode, wherein the electrode pair is disposed in the holder; and
a computer configured to set an analysis area on the two-dimensional image and analyze optical intensity in the analysis area,
wherein the electrode pair has a structure in which a tip of the first electrode on a bottom side of the holder extends more than a tip of the second electrode on the bottom side of the holder, and
wherein the electrode pair is positioned by bringing the tip of the second electrode into contact with a bottom face of the holder and by allowing a space between the tip of the first electrode and the bottom face of the holder, when the electrode pair is inserted into the holder.

### Claim 2 (depends on 1)

The system according to claim 1,
wherein the electrical stimulator is configured to repeatedly stimulate the sample with a cycle between 1 Hz to and 10 Hz.

### Claim 3 (depends on 1)

The system according to claim 1,
wherein the first electrode and the second electrode are plate-shaped electrodes.

### Claim 4 (depends on 1)

The system according to claim 1,
wherein the first electrode is disposed in a state in which the tip of the first electrode is separated by a predetermined distance from the bottom face of the holder.

### Claim 5 (depends on 1)

The system according to claim 1, further comprising
a light source configured to apply excitation light to the sample.

### Claim 6 (depends on 5)

The system according to claim 5,
wherein the light-guiding optical system is configured to guide the excitation light from the light source to the sample.