Hi all,
I wish to stain a kidney proximal tubule cell line for markers such as KIM-1, the SGLT's etc, but I require the whole surface of the wells in which the cells are cultured to remain flat and as such can't use coverslips.
If anyone has any protocols or suggestions re: ICC directly on cultured cells I would love to hear from you!
Many thanks
Tom Smith
Tom, the protocol is exactly the same with the exception that you do not need to place a coverslip in the well or transfer the coverslip to a slide after ICC. Other than that, I imagine you will have to make some changes to the way you capture images afterwards.
Will this be your first ICC? If so, I can give you some helpful tips.
Best
A 24 well plate would be ideal for minimal use of reagents/antibodies. But make sure you have microscope stage that fits with such plate.
I agree with Ricardo, it is the same protocol (except you won't have to fish the coverslips), however, you will not have slides to store (and storing lots of plates is not practical), also you will need an inverted microscope to take pictures (in case you use smaller plates; as 48/96 well plates).
Also as Prasanth said, Lab-teck chamber slides are very convenient, but they are expensive.
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