Cell evaluation method
View Patent ↗A cell evaluation method includes: a labeling step of labeling at least some of cells constituting a cell clump with a chemical substance; an image-capturing step of acquiring a plurality of slice images of the labeled cells, for at least part of the cell clump; an evaluating step of evaluating the characteristics of the cells on the basis of the plurality of slice images; a 3D-image creating step of creating a 3D image of at least part of the cell clump by subjecting the plurality of slice images to image processing; a center-of-gravity determining step of determining, from the 3D image, a center-of-gravity position of the cell clump; and an organizing step of organizing evaluation results obtained in the evaluating step, with respect to the distance from the center-of-gravity position.
1. A cell evaluation method comprising:
a labeling step of labeling at least some of cells constituting a cell clump with at least one chemical substance that produces fluorescence or luminescence;
an image-capturing step of acquiring a plurality of slice images of the cells that are labeled in the labeling step, for at least part of the cell clump;
an evaluating step of evaluating the characteristics of the cells on the basis of the plurality of slice images acquired in the image-capturing step;
a 3D-image creating step of creating a 3D image of at least part of the cell clump by subjecting the plurality of slice images acquired in the image-capturing step to image processing;
a center-of-gravity determining step of determining a center-of-gravity position of the cell clump from the 3D image of the cell clump created in the 3D-image creating step; and
an organizing step of organizing evaluation results obtained in the evaluating step, with respect to the distance from the center-of-gravity position determined in the center-of-gravity determining step.
2. The cell evaluation method according to claim 1 , wherein, in the organizing step, the evaluation results are organized with respect to the distance in a direction of gravitational force from the center-of-gravity position determined in the center-of-gravity determining step.
3. The cell evaluation method according to claim 1 , wherein, in the organizing step, at least part of the cell clump is divided into a plurality of regions according to the distance from the center-of-gravity position determined in the center-of-gravity determining step, and the evaluation results of the cells existing in each of the regions are organized for the region.
4. The cell evaluation method according to claim 1 , wherein, in the evaluating step, the amount of light produced in components detected in each of the cells is evaluated.
5. The cell evaluation method according to claim 4 , wherein the components are cell nuclei.
6. The cell evaluation method according to claim 1 , wherein, in the evaluating step, the number of components detected in the cells is evaluated.
7. The cell evaluation method according to claim 6 , wherein the components are cell nuclei.
8. The cell evaluation method according to claim 1 , wherein, in the evaluating step, the density of components detected in the cells is evaluated.
9. The cell evaluation method according to claim 8 , wherein the components are cell nuclei.
10. The cell evaluation method according to claim 1 ,
wherein, in the labeling step, a first chemical substance that can specifically label cell nuclei and a second chemical substance that can specifically label cell nuclei of dead cells and that produces light having a different wavelength from that produced by the first chemical substance are used to label the cell nuclei; and
in the evaluating step, the ratio of the dead cells is evaluated.
11. The cell evaluation method according to claim 1 ,
wherein, in the labeling step, a fluorescent protein whose fluorescence wavelength changes according to a phase of a cell cycle is used to label the cells; and
in the evaluating step, the ratio of the phase of the cell cycle is evaluated.
12. The cell evaluation method according to claim 1 , wherein components detected in each of the cells are cell cytoplasm.
13. The cell evaluation method according to claim 1 ,
wherein, in the labeling step, a chemical substance that can specifically label cell nuclei is used to label the cell nuclei; and
in the evaluating step, only micronuclei that have volumes less than a predetermined threshold are extracted from a 3D image of cell nuclei and the micronuclei, acquired in the 3D-image creating step, and an incidence of the micronuclei is evaluated.
14. The cell evaluation method according to claim 1 ,
wherein, in the labeling step, a chemical substance that can specifically label cell nuclei is used to label the cell nuclei;
in the evaluating step, fragmented cell nuclei that have volumes less than a predetermined threshold are extracted from a 3D image of the fragmented cell nuclei acquired in the 3D-image creating step, and a fragmentation percentage of the cell nuclei is evaluated.
15. The cell evaluation method according to claim 1 ,
wherein, in the labeling step, a chemical substance that can specifically label cell membranes is used to label the cell membranes; and
in the evaluating step, an internal volume of cell membranes is calculated from a 3D image of the cell membranes that is acquired in the 3D-image creating step, thereby evaluating a volume of the cells.
16. The cell evaluation method according to claim 1 , wherein, in the center-of-gravity determining step, the center-of-gravity position is determined on the basis of, among cross sections of a 3D image of the whole cell clump along a plurality of planes parallel to each other, a cross section of the cell clump in the plane that has the maximum cross-section area of the cell clump.
17. The cell evaluation method according to claim 1 , wherein, in the center-of-gravity determining step, the center-of-gravity position of the cell clump is estimated on the basis of the distribution of components of each of the cells in the 3D image.