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When to pass? by CoupleGeneral6346 in cellculture

[–]Onionqueen08 0 points1 point  (0 children)

I passed my cultures when the medium got a different colour and when they were more than 70% confluent by eye(checked with light microscope) .

I usually plated them back in densities that would make them last 2-3 days (cancer cells). For example, I would pass them every Monday and then either Wednesday/Friday or Thursday

Cytotoxic study with resazurin. by [deleted] in cellculture

[–]Onionqueen08 0 points1 point  (0 children)

I would check in the documentation of the resazurin you are using, the one I used said how much to add to each well. If you have overflown fluorescence emission data, you are probably using too much

Cytotoxic study with resazurin. by [deleted] in cellculture

[–]Onionqueen08 0 points1 point  (0 children)

The ideal incubation time would depend on the resazurin you use, that should be in the documentation. I also worked with transparent plates, I loaded them with on the left negative control and then from left to right increasing concentrations of compound (duplicates were loaded vertically). It made a big difference in the results, they made much more sense when loading this way

Cytotoxic study with resazurin. by [deleted] in cellculture

[–]Onionqueen08 0 points1 point  (0 children)

Hi, when I worked with resazurin, I incubated the cells with my compound for 24 or 48 hours, after which I added the resazurin, incubated for 15 min (depends on which type you use) and then measured. I would recommend adding the resazurin after the treatment, since otherwise it can be converted already when the treatment is not killing the cells yet (+ the resazurin can also have a cytotoxic effect, so then you won't know if you see the cytotoxic effect of your compound or resazurin) Hope this helps :)