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?