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 cell based on its position on one or more surfaces or in one or more containers, wherein the selecting is not based on whether the cell comprises an exogenous label or an expressed reporter and wherein the selecting is performed using an imaging technique and an automated image analysis process; and
b) photoablating at least 80% of five or more cells on each of the one or more surfaces or in each of the one or more containers, wherein at least 95% of the one or more surfaces or containers contain only one viable cell after the photoablating is performed, wherein the cell selected in (a) is not photoablated.
2. The method of claim 1 , wherein b) comprises photoablating all but a first cell of the five or more cells on each of the one or more surfaces or in each of the one or more containers, wherein the first cell is the cell selected in (a).
3. The method of claim 2 , further comprising growing a clonal population of the first cell after the photoablating is performed, thereby generating a clonal population of cells.
4. The method of claim 3 , further comprising performing an assay on the clonal population of cells.
5. The method of claim 3 , further comprising photodetaching one or more cells of the clonal population of cells from the one or more surfaces or one or more containers thereby obtaining one or more photodetached cells.
6. The method of claim 5 , further comprising performing an assay on the one or more photodetached cells.
7. The method of claim 5 , further comprising growing the one or more photodetached cells.
8. The method of claim 2 , wherein the five or more cells comprise about 10 to about 15 cells.
9. The method of claim 2 , wherein the photoablating occurs at a rate of at least 60 cells per minute.
10. The method of claim 2 , wherein the photoablating occurs with an efficiency of greater than 99%.
11. The method of claim 2 , wherein the photoablating occurs using light in the wavelength range of 1440 nm to 1450 nm.
12. The method of claim 1 , wherein the selecting is based on a proximity of the first cell to a center of the one or more surfaces or containers, a size of the first cell, a morphology of the first cell, a phenotype of the first cell, a development stage of the first cell, or any combination thereof.
13. The method of claim 1 , wherein at least 98% of the one or more surfaces or containers contain only one viable cell after the photoablating is performed.
14. 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.
15. The method of claim 1 , wherein the imaging technique comprises bright-field imaging, dark-field imaging, phase contrast imaging or any combination thereof.
16. The method of claim 1 , wherein the selecting is based on a proximity of the cell to a center of the one or more surfaces or containers.
17. The method of claim 1 , wherein the selecting is based on a size of the cell.
18. The method of claim 1 , wherein the selecting is based on a morphology of the cell.
19. The method of claim 1 , wherein the selecting is based on a phenotype of the cell.
20. The method of claim 1 , wherein the selecting is based on a development stage of the cell.