Live cell analysis apparatus, method for analyzing live cells, system for analyzing live cells and non-transitory data storage medium

Patent metadata

Field	Value
Publication number	US-10168314-B2
Application number	US-201515119684-A
Country	US
Kind code	B2
Filing date	Feb 13, 2015
Priority date	Mar 3, 2014
Publication date	Jan 1, 2019
Grant date	Jan 1, 2019

Abstract

Described are methods and structures associated with live cell analysis apparatus. A live cell analysis apparatus may include at least one data processor that is configured to process a sequence of images of live cells to determine motion of one or more live cells. The motion may be in response to applied electrical stimulations. The apparatus and methods may be used to determine a refractory period for cardiomyocytes from the determined motion.

First claim

The invention claimed is:

1. A live cell analysis apparatus, comprising:
an electro stimulator configured to apply first electrical stimulations to at least one live cell;
a microscope configured to capture a sequence of images of the at least one live cell; and
at least one data processor configured to:
process the sequence of images to determine motion of the at least one live cell that is responsive to the applied first electrical stimulations; and
determine a refractory period for the at least one live cell based on the determined motion.

2. The live cell analysis apparatus of claim 1 , wherein the at least one live cell corresponds to at least one cardiomyocyte.

3. The live cell analysis apparatus of claim 1 , wherein:
the electro stimulator is further configured to apply the first electrical stimulations for pacing a plurality of cardiomyocytes, and
the at least one data processor is further configured to process the sequence of images of the plurality of cardiomyocytes to determine a first phase delay between an initial contraction of the plurality of cardiomyocytes and a relaxation of the plurality of cardiomyocytes.

4. The live cell analysis apparatus of claim 3 , wherein the electro stimulator is further configured to apply second electrical stimulations to the at least one live cell, delayed from the first electrical stimulations by a selectively varied second phase delay.

5. The live cell analysis apparatus of claim 4 , wherein the selectively varied second phase delay is initially set equivalent to the first phase delay.

6. The live cell analysis apparatus of claim 4 , wherein the at least one data processor is further configured to determine a refractory period for the plurality of cardiomyocytes based on at least the determined motion and a value of the selectively varied second phase delay, and wherein the plurality of cardiomyocytes comprise the at least one live cell.

7. A method for analyzing live cells, the method comprising:
applying first electrical stimulations to at least one live cell;
capturing a sequence of images, with a microscope, of the at least one live cell;
determining by at least one data processor:
motion of the at least one live cell based on the sequence of images, wherein the motion is based on the applied first electrical stimulations; and
a refractory period for at least one live cell based on the determined motion.

8. The method of claim 7 , wherein the at least one live cell correspond to at least one cardiomyocyte.

9. The method of claim 8 , further comprising:
determining a first refractory period for a plurality of first cardiomyocytes of the plurality of cardiomyocytes based on first determined motion data, wherein the plurality of first cardiomyocytes are not treated with a pharmaceutical drug;
determining a second refractory period for a plurality of second cardiomyocytes of the plurality of cardiomyocytes based on second determined motion data, wherein the plurality of second cardiomyocytes are treated with the pharmaceutical drug; and
comparing the first refractory period and the second refractory period.

10. The method of claim 7 , further comprising:
applying the first electrical stimulations for pacing a plurality of cardiomyocytes; and
determining, by the at least one data processor, a first phase delay between an initial contraction of the plurality of cardiomyocytes and a relaxation of the plurality of cardiomyocytes.

11. The method of claim 10 , further comprising applying second electrical stimulations to the at least one live cell, delayed from the first electrical stimulations by a selectively varied second phase delay.

12. The method of claim 11 , further comprising initially setting the selectively varied second phase delay equivalent to the first phase delay.

13. The method of claim 11 , further comprising calculating a refractory period for the plurality of cardiomyocytes based on at least the determined motion and a value of the selectively varied second phase delay, wherein the plurality of cardiomyocytes comprise the at least one live cell.

14. The method of claim 13 , wherein calculating the refractory period comprises:
producing at least one waveform representative of the determined motion; and
analyzing the at least one waveform to calculate the refractory period.

15. The method of claim 13 , wherein calculating the refractory period for the plurality of cardiomyocytes comprises changing the selectively varied second phase delay until one of:
the plurality of cardiomyocytes no longer respond to the second electrical stimulations, or
the plurality of cardiomyocytes respond to the second electrical stimulations.

16. The method of claim 13 , further comprising determining one of a propagation speed or a propagation direction based on the determined motion.

17. The method of claim 16 , wherein one of the propagation speed or the propagation direction, and the refractory period are determined from the sequence of images.

18. The method of claim 13 , further comprising:
changing a first position of at least the first electrical stimulations to at least one second position;
determining a third refractory period corresponding to the at least one second position; and
determining a homogeneity in response of the plurality of cardiomyocytes from the determined third refractory period associated with the at least one second position.

19. A system to analyze live cells, the system comprising:
an electro stimulator configured to apply first electrical stimulations to at least one live cardiomyocyte;
a microscope configured to capture a sequence of images of the at least one live cardiomyocyte; and
at least one data processor connected to the microscope and the at least one data processor configured to:
process the sequence of images to determine motion of the at least one live cardiomyocyte, wherein the motion is based on the applied first electrical stimulations; and
determine a refractory period of the at least one live cardiomyocyte.

20. A non-transitory computer-readable medium having stored thereon computer-executable instructions that, when executed by at least one data processor of a live cell analysis apparatus, cause the live cell analysis apparatus to:
apply electrical stimulations to at least one live cardiomyocyte;
capture a sequence of images, with a microscope, of the at least one live cardiomyocyte;
determine from the sequence of images, by the at least one data processor, motion of the at least one live cardiomyocyte, wherein the motion is based on the applied electrical stimulations; and
calculate a refractory period for the at least one live cardiomyocyte based on the determined motion.

21. An apparatus for live cell analysis, comprising:
a controller configured to control electrical stimulations applied to at least one live cell;
an image processor configured to receive a sequence of images of the at least one live cell from a microscope; and
at least one data processor configured to:
process the sequence of images to determine motion of the at least one live cell, wherein the motion is based on the applied electrical stimulations, and
determine a refractory period for a plurality of cardiomyocytes based on the determined motion, wherein the at least one live cell is at least one cardiomyocyte of the plurality of cardiomyocytes.

Assignees

• Sony Corp

Inventors

• Takahashi Kazuya
• Hayakawa Tomohiro
• Tatsuta Hirokazu
• Matsui Eriko

Classifications

• G01N33/48728

  Investigating individual cells, e.g. by patch clamp, voltage clamp (investigating individual particles in general G01N15/10) · CPC title

• G01N33/48792 Primary

  Data management, e.g. communication with processing unit (for in vivo diagnostics A61B5/0002; transmission systems for measured values G08C) · CPC title

• G01N33/48707 Primary

  by electrical means (G01N33/49, G01N33/493 take precedence) · CPC title