Cell culture laser photoablation
Methods and systems for preparing clonal cell populations are described. In some instances the disclosed methods comprise: a) identifying and selecting a cell based on its position on a surface or in a container, where the selection is not based on whether the cell comprises an exogenous label or an expressed reporter; b) photoablating all non-selected cells on the surface or in the container; and c) growing a clonal population of the selected cell.
1. A method comprising:
a) selecting a first cell on one or more surfaces or in one or more containers, wherein the selecting is not based on whether the first cell comprises an exogenous label or an expressed reporter and wherein the selecting is performed without reference to cell-specific features; and
b) photoablating at least 80% of the cells on each of the one or more surfaces or in each of the one or more containers, wherein at least 90% of the one or more surfaces or containers contain only one viable cell after the photoablation is performed, wherein the first cell selected in (a) is not photoabalated.
2. The method of claim 1 , wherein at least 95% of the one or more surfaces or containers contain only one viable cell after the photoablation is performed.
3. The method of claim 1 , wherein at least 98% of the one or more surfaces or containers contain only one viable cell after the photoablation is performed.
4. The method of claim 1 , wherein the one or more surfaces or containers comprise at least 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 surfaces or containers.
5. The method of claim 1 , wherein the first cell is selected using an imaging technique.
6. The method of claim 5 , wherein the imaging technique comprises bright-field imaging, dark-field imaging, phase contrast imaging or any combination thereof.
7. The method of claim 1 , wherein the selecting is based on position of the first cell on one or more surfaces or in one or more containers.
8. The method of claim 7 , wherein the selecting is based on proximity of the first cell to a center of the one or more surfaces or containers.
9. The method of claim 1 , wherein the selecting is based on size of the first cell.
10. The method of claim 1 , wherein the selecting is based on a morphology of the first cell.
11. The method of claim 1 , wherein the selecting is based on a phenotype of the first cell.
12. The method of claim 1 , wherein the selecting is based on a development stage of the first cell.
13. The method of claim 1 , wherein cells are photoablated at a rate in the range of 10 cells per minute to 200 cells per minute.
14. The method of claim 1 , wherein cells are photoablated using light in the wavelength range of 220 nm to 1500 nm.
15. The method of claim 14 , wherein cells are photoablated using light in the wavelength range of 1440 nm to 1450 nm.
16. The method of claim 1 , further comprising growing a clonal population of the first cell after the photoablation is performed.
17. The method of claim 16 , further comprising performing an assay on the clonal population of the first cell.
18. The method of claim 16 , further comprising photodetaching one or more cells of the clonal population of the first cell from the one or more surfaces or one or more containers thereby obtaining one or more photodetached cells.
19. The method of claim 18 , further comprising performing an assay on the one or more photodetached cells.
20. The method of claim 18 , further comprising growing the one or more photodetached cells.