I have difficulty every time whenever I subculture C6/36 Cell line and vero cell line.
Can anyone suggest how to spread cells evenly in growth media.
1. Do subculturing or splitting cells in low dilutions such as 1:3 to 1:10. 1:3 is best for doing plaque assays etc, which require evenly distributed and well-spread cells for accurate results. If you split in higher dilutions, each cell forms a colony and it takes time for colonies to touch each other and meanwhile the middle of your colony becomes so dense, which is not good.
2. You need to mix the cell suspension properly during splitting. you need to make sure that there are no cell clumps which form dense areas in the plate.
3. If your incubator is not leveled correctly, keep the cells on the flat surface for 5 min before keeping in the incubator.
4. Mix properly after addition of the concentrated culture into the media. Avoid swirling movement of the plate which produce centripetal forces which increase the cell density in the middle of the plate.
Goodluck.
I would add to Ramachandramouli's answer that instead of swirling in a circular motion, swirl the plated cells in a figure 8 motion or side-to-side then up-and-down. This will mix the cells in the plate without pushing the cells to the outside of the dish. Cheers.
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