I did my PhD in the Department of Chemical Engineering at Imperial College London. My research focused on mathematical modeling of the cell cycle in leukemia and involved experiments with cell lines. During that time, I had to count cells with a hemocytometer so often to track growth that I got tired and decided to build an app, HemocyTap, and share my knowledge on the topic here to help as many people as possible.

Your cells are in suspension already so there is no need to take any extra action before doing a cell count. Just make sure you mix properly before taking a sample, and proceed to “How to count cells?”. Depending on the color of your medium, you might want to do two things:

  • If the colour of your medium has changed to orange/yellowish:

you have plenty of metabolites and your cells won’t like that. You definitely want to centrifuge your cells, aspirate the old medium and resuspend in completely new medium, according to the recommended seeding density. Transfer them to new flasks and place in the incubator.

  • If the colour of your medium is still pinkish:

you can dilute your cells down with new medium to the recommended seeding density and split into more flasks, according to cell count.

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