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I'm trying to add on the order of 10-50 mammalian cells to a mixture of other cells (order of 1e6 cells). What is the best way to do this?

(edit) I guess this wasn't clear. I would really like to emphasize how to sort 10-50 cells to give me an precise (not necessarily accurate but that would be nice too) method of getting a desired number of cells.

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If you are you working with primary cell cultures or cell lines may be you could do this when cells are in suspension, after tripsinization, mix them well with a 5mL serologial pipete in a falcon 50mL tube and plate to a 100mm2 with a final volume of 7mL. And finaly draw an eight with the plate in the hood more than 3 times to mix them well.

Edit: For the addition of 10-50 cells, count these cells with trypan blue, and do a 1:100 dilution, for example if you have 1e6 cells in 10mL you could take 0.01 mL (10 microliters) to get 1, 000 cells

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And then you add the 0.01mL into 1mL of cell media to get 1 cell per 1 microliter, and then if you wanna get 10 cells, take 10 microliters, and so on

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  • $\begingroup$ So what about the getting the 50 cells part? $\endgroup$
    – bobthejoe
    Commented Jun 19, 2012 at 7:20
  • $\begingroup$ Check the edition of the answer $\endgroup$
    – friveroll
    Commented Jun 19, 2012 at 16:33
  • $\begingroup$ How do you avoid stochastic effects? $\endgroup$
    – bobthejoe
    Commented Jun 19, 2012 at 20:16
  • $\begingroup$ You can't avoid random effects in biology, because biological variability, do a power analysis to ensure you use the correct numer of samples or repeats in your experiment biology.stackexchange.com/questions/963/…. My answer is a standard method to work with cell cultures, you can also use cell sorting, to be more precise on your cell count. And try bioforum (a community of methods), to get another solutions $\endgroup$
    – friveroll
    Commented Jun 20, 2012 at 15:37

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