Hi, I need to know about cell counting for time point experiments-

I am working on Raw 264.7 cells which has 11 hours doubling time.

I use to seed 1 Million cells in six well plate which has capacity of ~1.2 million cells but they become 2 million before I done with infection.

I can control this by seeding the cells just 5-6 hrs before infection since Raw cells usually take 5-6 hrs to get attach.

But how to control time points?

I need to take out samples at 6,12,18 and 24 hrs.

I use to infect at MOI of 1:10, so I use to add same amount at one time only but for my 24 hrs exp. cells become more so I feel infection got diluted? How to control this?

One more thing, to handle this i also seed 1 million cells in 60 mm plate, but I am worried since cells are less and surface area is more, is it going to affect infection?

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