HI all, can anyone tells me if we want to infect any cell line with bacteria so can we seed cells up to maximum amount we can seed in an dish or less?
like in six well i use to seed 1 million o/n in ab free media and then i infect in morning, taking samples at 6 hour interval til 24 hrs.so, obviously at 24hrs comparison to 6 hrs there will be more growth?
i need to know is it good practice to count cell and then plate on respective dish or first grow itself on the same?
obviously we can seed same amount but if my exp involves time kinetics study, so how much this going to vary my experiments??
Hi Shilpi,
It depends on the type of work you intend to do. If you want to do an adhesion or invasion assay, which requires you to take photos under a microscope, then you need not to wait till the cell becomes 100% confluent. You only be needing 60-70% confluence. But if you're doing an assay which requires you to plate count the bacteria at the end or if you want to study the signaling pathways, you need to seed more cells and/or to wait till it gets more than 90% confluent.
And yes, you do need to maintain the cell numbers strictly in experiments or else it will hamper the results. Signaling study will get hampered if you seed and grow varying numbers of cells every time.
Hope this will help.
Regards.
Hi Shilpi,
if you include the appropriate control for every timepoint you should be fine. Just compare the T0 control to T0 infected, T1 control to T1 infected and so on.
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