Hi All,
When I subculture the cell from one vessels (flask) to new multiple vessels (six-well, petri dishs). I often have one or two culure vessel that has less or more cells than the others.
I subculture with trypsin, wait all detach, and inactivate with culture media, then I have gently resuspend the cell solution about 20 times and checked there is no cell clump by microscope before passage equally volume of the cell into new vessels. The cell are distributed equally on culture suface.
Do you have any tip to subculture equally number of cells from one vessel into multiple new vessels? I am thinking maybe using low setting vortex, but never try.
Cheerio,
Actually this is exactly what you are supposed to do. However be aware that cell culture is not always super accurate, so you cannot expect totally equal distribution. That said, you can get pretty close to equal numbers. I would not vortex, resuspending is usually fine. It could be that you are too slow in transferring the cells into the new vessel, i.e. meaning that some cells have already sedimented at the bottom of your cell suspension when you reached your last vessel?
In case of 5 new vessels, you could also resuspend your cells in 20ml of medium and then take up the complete 20ml of cell suspension with a pipette (25ml pipette is usually used). Then transfer 4ml from that pipette into each new vessel. This takes about 30sec with some experience in cell culture. Then just add up to your desired volume with medium in your new vessels, et voila!
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