Cell detection device and cell detection method

Patent metadata

Field	Value
Publication number	US-10801007-B2
Application number	US-201916591670-A
Country	US
Kind code	B2
Filing date	Oct 3, 2019
Priority date	May 20, 2015
Publication date	Oct 13, 2020
Grant date	Oct 13, 2020

Abstract

A cell detection device and a cell detection method is provided with which it is possible to efficiently acquire images of cells to be measured. Cell detection device comprises flow cell through which a measurement specimen that contains particles is caused to flow, particle detector for detecting the particles in the measurement specimen supplied to flow cell, particle sorter for sorting particles that satisfy a detection condition and other particles on the basis of the result of detection performed by particle detector, specimen supply part for supplying, to flow cell, an image-capture specimen that includes detection-condition-satisfying particles that have been sorted by particle sorter, and particle-image-capture part for capturing images of the particles in the sorted image-capture specimen supplied to flow cell.

First claim

The invention claimed is:

1. A sample analysis method, comprising:
introducing a measurement specimen to a flow cell through an inlet;
sensing cells in the measurement specimen at a first position in the flow cell;
directing target cells among the cells sensed at the first position, to a recirculation flow path at a second position that is downstream of the first position, wherein the recirculation flow path connects the second position and the inlet;
supplying the target cells to the flow cell via the recirculation flow path; and
capturing images of the target cells in the flow cell, wherein
the recirculation flow path comprises an intermediate reservoir adapted to increase a concentration of the target cells.

2. The method according to claim 1 , wherein the intermediate reservoir comprises an elutriator rotor, a centrifuge or a filter.

3. The method according to claim 1 , wherein the sensing comprises reading an optical characteristics of individual cells.

4. The method according to claim 3 , wherein the reading comprises:
irradiating light onto the cells in the measurement specimen passing the first position; and
detecting a scattered light and a fluorescence emitted from the individual cells.

5. The method according to claim 1 , wherein
a width of a flow path in the flow cell in a first direction perpendicular to a direction in which the measurement specimen flows is greater than a width of the flow path in a second direction perpendicular to the first direction; and
the target cells are imaged, facing to the second direction.

6. The method according to claim 1 , wherein
an imaging device is coupled to the flow cell and controlled to capture images of individual cells at the first position for sensing cells, and
the imaging device is controlled to capture images of the target cells supplied from the recirculation flow path.

7. The method according to claim 1 , wherein
the measurement specimen is introduced to the flow cell with a first flow speed, and
the target cells are supplied from the recirculation flow path to the flow cell with a second flow speed that is reduced than the first flow speed.

8. The method according to claim 1 , wherein
the flow cell is connected to a reservoir storing a sheath liquid, and
a flow speed of the target cells in the flow cell is reduced by increasing a volume of the sheath liquid supplied to the flow cell.

9. The method according to claim 1 , further comprising applying an ultra sound to the flow cell so that the target cells flowing in the flow cell are aligned.

10. The method according to claim 1 , further comprising collecting the target cells at a downstream of the flow cell.

11. The method according to claim 1 , wherein the target cells comprise circulating tumor cell (CTC), Circulating Endothelial Cell (CEC), Endothelial Progenitor Cell (EPC), Mesenchymal Stem Cell (MSC), Hematopoietic Stem Cell (HSC) or antibody-specific T cell.

12. The method according to claim 1 , further comprising mixing a biological sample collected from a patient and a reagent to prepare a measurement specimen.

13. The method according to claim 12 , wherein the reagent contains a labeled antibody which binds to a specific site of the target cell.

14. The method according to claim 12 , wherein the reagent contains a nucleic acid probe which hybridizes to a specific site of a nucleic acid in the target cell.

15. The method according to claim 12 , wherein the biological sample comprises a peripheral blood and the reagent contains a hemolytic agent to hemolyze red blood cells.

16. A sample analysis method, comprising:
introducing a measurement specimen to a flow cell through an inlet;
irradiating light onto the flow cell and reading an optical response from individual particles in the measurement specimen flowing inside the flow cell;
actuating a particle sorter that is coupled to a downstream of the flow cell, in response to an optical response indicative of a target cell, to change a direction of the target cell;
re-introducing the target cell, through a recirculation flow path connecting a downstream of the particle sorter and the inlet, to the flow cell; and
capturing, by an imaging device, an image of the target cell in the flow cell.

17. The method according to claim 16 , wherein
the target cell presents as plural target cells in the measurement specimen, and
the plural target cells are accumulated in the recirculation flow path and then introduced to the flow cell.

18. A sample analyzer, comprising:
a flow cell to receive a measurement specimen flowing through an inlet;
a particle detector operable to sense cells in the measurement specimen at a first position in the flow cell;
a particle sorter, positioned at a second position downstream to the first position, operable to change a direction of target cells among the cells sensed at the first position;
a recirculation flow path connecting the inlet and a downstream of the particle sorter to receive the target cells directed by the particle sorter and to supply the target cells to the flow cell; and
an imaging device coupled to the flow cell to capture images of the target cells, wherein
the recirculation flow path comprises an intermediate reservoir adapted to increase a concentration of the target cells.

19. A sample analysis method, comprising:
introducing a measurement specimen to a flow cell through an inlet;
sensing cells in the measurement specimen at a first position in the flow cell;
directing target cells among the cells sensed at the first position, to a recirculation flow path at a second position that is downstream of the first position, wherein the recirculation flow path connects the second position and the inlet;
supplying the target cells to the flow cell via the recirculation flow path; and capturing images of the target cells in the flow cell, wherein
an imaging device is coupled to the flow cell and controlled to capture images of individual cells at the first position for sensing cells, and
the imaging device is controlled to capture images of the target cells supplied from the recirculation flow path.

20. A sample analysis method, comprising:
introducing a measurement specimen to a flow cell through an inlet;
sensing cells in the measurement specimen at a first position in the flow cell;
directing target cells among the cells sensed at the first position, to a recirculation flow path at a second position that is downstream of the first position, wherein the recirculation flow path connects the second position and the inlet;
supplying the target cells to the flow cell via the recirculation flow path; and capturing images of the target cells in the flow cell, wherein
the measurement specimen is introduced to the flow cell with a first flow speed, and
the target cells are supplied from the recirculation flow path to the flow cell with a second flow speed that is reduced than the first flow speed.

Assignees

• Sysmex Corp

Inventors

• Tabata Seiichiro
• Yanagida Masatoshi

Classifications

• G01N1/34

  Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)} · CPC title

• G01N1/2813 Primary

  Producing thin layers of samples on a substrate, e.g. smearing, spinning-on (G01N1/30 takes precedence) · CPC title

• G01N1/40 Primary

  Concentrating samples · CPC title

• C12M47/04

  Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04) · CPC title

• G01N15/1436 Primary

  the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell · CPC title