Cell-sorting method and associated device

Patent metadata

Field	Value
Publication number	US-10400208-B2
Application number	US-201414907260-A
Country	US
Kind code	B2
Filing date	Jul 21, 2014
Priority date	Jul 23, 2013
Publication date	Sep 3, 2019
Grant date	Sep 3, 2019

Abstract

A method of discriminating a living cell from a dead cell, and a device for implementing the method, the method including: using a lens-free imaging device to acquire a diffraction figure corresponding to a cell; and determining a light intensity on a central area of an elementary diffraction figure associated with the cell. It can thus be determined if the studied cell is a living cell or a dead cell.

First claim

The invention claimed is:

1. A method of discriminating a living cell from a dead cell implemented using a lens-free imaging device including a light source and a matrix photodetector, the method comprising:
using the light source to illuminate a sample of a liquid solution containing cells;
arranging the matrix photodetector facing the light source, and acquiring a global diffraction figure of the sample, the global diffraction figure comprising a plurality of elementary diffraction figures each associated with a cell, each elementary diffraction figure having a succession of closed concentric curves; and
classifying a cell in one of a living cells category or a dead cells category, depending on a light intensity on a central area of an elementary diffraction figure associated with the cell,
wherein the central area is inscribed in an area delimited by a smallest diffraction ring of the elementary diffraction figure.

2. A method according to claim 1 , further comprising:
determining a value of a numeric indicator representative of the light intensity in the central area;
comparing the value of the numeric indicator with a predetermined threshold value; and
depending on a result of this comparison, classifying the cell in one of the living cells category or the dead cells category.

3. A method according to claim 2 , wherein the numeric indicator is a maximum intensity or a mean intensity or a peak-to-peak intensity value.

4. A method according to claim 3 , wherein the predetermined threshold value is a reference intensity, and:
a numeric indicator value lower than the reference intensity corresponds to a living cell; and
a numeric indicator value higher than the reference intensity corresponds to a dead cell.

5. A method according to claim 1 , used to perform in situ monitoring of a cell treatment in a bioreactor.

6. A method of discriminating a living cell from a dead cell implemented using a lens-free imaging device including a light source and a matrix photodetector, the method comprising:
using the light source to illuminate a sample of a liquid solution containing cells,
arranging the matrix photodetector facing the light source, and acquiring a global diffraction figure of the sample, the global diffraction figure comprising a plurality of elementary diffraction figures each associated with a cell, each elementary diffraction figure having a succession of closed concentric curves; and
classifying a cell in one of a living cells category or a dead cells category, depending on a light intensity on a central area of an elementary diffraction figure associated with the cell,
wherein the global diffraction figure of the sample is acquired using a matrix photodetector placed facing the light source, no magnification device being placed between the sample and the matrix photodetector.

7. A device for discriminating a living cell from a dead cell comprising:
a light source, arranged to illuminate a sample of a liquid solution comprising cells; and
a matrix photodetector placed facing the light source, forming a lens-free imaging device with the light source, and arranged to acquire a global diffraction figure of the sample, the global diffraction figure comprising plural elementary diffraction figures each of which is associated with a cell, each elementary diffraction figure having a succession of closed concentric curves;
a calculation means configured to:
receive the global diffraction figure as an input;
determine the light intensity on a central area of an elementary diffraction figure associated with a cell;
provide as an output a classification of the cell as a living cell or a dead cell;
wherein no magnification device is placed between the sample and the matrix photodetector;
a first sealed housing, holding the light source and including a first transparent window between the light source and the matrix photodetector; and
a second sealed housing, holding the matrix photodetector, and including a second transparent window between the first transparent window and the matrix photodetector;
wherein a distance between the first transparent window and the second transparent window is variable;
the first sealed housing, the light source, the second sealed housing, and the matrix photodetector together forming an immersible probe.

8. A device according to claim 7 , wherein the distance between the first transparent window and the second transparent window is variable in time, the first transparent window and the second transparent window defining two planes parallel to each other, and the device further comprising means for displacing the second transparent window relative to the first transparent window.

9. A device according to claim 8 , wherein the means for displacing the second transparent window relative to the first transparent window includes a rack driven in translation by a gear, and fixed relative to the second sealed housing.

10. A device according to claim 7 , wherein the distance between the first transparent window and the second transparent window is variable in space.

11. A device according to claim 10 , wherein the first transparent window and the second transparent window define two planes inclined relative to each other, such that the distance between the first transparent window and the second transparent window depends on a location on the second transparent window.

12. A device for discriminating a living cell from a dead cell comprising:
a light source, arranged to illuminate a sample of a liquid solution comprising cells; and
a matrix photodetector placed facing the light source, forming a lens-free imaging device with the light source, and arranged to acquire a global diffraction figure of the sample, the global diffraction figure comprising plural elementary diffraction figures each of which is associated with a cell, each elementary diffraction figure having a succession of closed concentric curves;
a calculation means configured to:
receive the global diffraction figure as an input;
determine the light intensity on a central area of an elementary diffraction figure associated with a cell;
provide as an output a classification of the cell as a living cell or a dead cell; and
wherein no magnification device is placed between the sample and the matrix photodetector, and
wherein the calculation means determines the light intensity on a central area of an elementary diffraction figure associated with a cell, the central area being inscribed in an area delimited by a smallest diffraction ring in the elementary diffraction figure.

Assignees

• Commissariat Energie Atomique

Inventors

• Allier Cedric

Classifications

• G03H2222/34

  Multiple light sources · CPC title

• G03H2001/0447

  In-line recording arrangement · CPC title

• G03H2001/0033

  in hologrammetry for measuring or analysing · CPC title

• G03H1/0465

  Particular recording light; Beam shape or geometry (G03H1/06 takes precedence) · CPC title

• G03H1/0443

  Digital holography, i.e. recording holograms with digital recording means (holobject computation G03H1/0866) · CPC title