How much T.C media should I put in the 24 well plate or the 96 well plate or the roux bottle? Should I put the microlites of cells in each vessel?
I am working on BHK -21 cells.
We use 200ul on 96 well plate, but only it is only good for a few days. On 24 well, 1ml? 0.5ml? I'd have to go in hood. We use 3ml in 6 well plate. Fill 24 well about 1/2 full. Make sure cells are well covered. Fewer numbers of cells can go longer. If you are doing a MTT test or IC50, you need to know how many cells per well and make sure your control doesn't "starve" and yet you still have enough cells to give good results. Also, the assays you will be sensitive to number of cells you use, so you have to have a "min." for each test (and that differs). On an MTT assay, for fast growing cell lines, you only need 20,000 per well but for some colon cancer HCT116 for MTT you want, 50,000/well. Some newer methods use very few cells per well, only 5,000/well.
So try increments of 5,000/well of your cells (i.e. 5,000, 10,000 etc.) in both 96 well and you 24 well, and just watch them and record. Write down when the media turns basic for each #, and when cell start to get hungry (sort of shrink and look bad, or grainy). This gives a good ball park. I don't know if you if you will be doing 24hr, 48hr, 72hr, or 96hr test and of what assay. But by doing this you'll know how many cell/well and when you have to feed them. Then for each test you can better plan out length of time and number of cells you'll need.
Also use exponential growing cells, not confluent, unless you are studying effects of confluence.
- Badges
- Science method