I have successfully established MIN6 cell Culture and now planing to do further experiments with cell viability assay and cell cytotoxicity assay. For this I am asking for guidance on protocols. How can we calculate the density of cell or density per well, how can we measure the seeding quantity and cell counting etc.
Hi Shiv,
Please follow this link for some useful numbers in cell culture - http://tools.lifetechnologies.com/downloads/Useful_Numbers_Y14472_Useful_Nmbrs.pdf
Apart from that you should also consider the factors like growth rate/doubling time of the cells that you are working on as well as the type of assay/analysis you are going to do on the cells; before seeding the cells. You should use trypan blue along with haemocytometer for cell counting and most of the assays should be done when your cells are 80-90% confluent.
Hope this helps! Good luck.
Best,
Abhijit
Already you got the answer, as we are following it. It will be easy for you while seeding the cells.
you also have to consider at what stage of the culture you will perform your test, .
I usually begin to assess acute (3-16 hrs) cytotoxicity, viability and so on when the cell monolayers reach the semiconfluence , ie 70-75% of the area of culture.
In this way at the end of experimetal time the cells are been able to proliferate,
Harita Parikh thanks for your guidance...can you provide me the protocol for cell titration...and other information related to efficient performance of the....cell titration assay...
Very very thank you to Anna M Bassi, utkarsh reddy addi, AL-namnam Nisreen a for your valuable opinion and support.....
I shall repeat the question, my question is "How can we measure cell density per well ", that is at the end of the experiment. One way is to collect cells from several wells, count and then calculate for single well, but how can we apply statistics then. please help if some one know.
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