Number of cells to start with (Using Suspension Cells)

[foxtr...@gmail.com]

Hi,
I am very new to RNA FISH and we are trying to setup the method in the lab. Because we are working with suspension cells, I wonder what is a good cell number to start with for fixing and labeling and also what amount of cells to ultimately be put on to the glass slide for imaging?

Thanks alot for your help!

[stevens...@gmail.com]

I usually work with at least 60,000 cells, sometimes more, because the protocol tends to reduce a huge chunk of your cells after the permeabilization step

[Arjun Raj]

Suspension is tricky because of cell loss during pelleting. I recommend adding 0.5% Triton to help then slide down, lo-bind tubes, and a swinging bucket rotor.

[stevens...@gmail.com]

Where do you think the adding of 0.5% Triton would help? after cells have been permeabilised?
Thanks
Steven