I have read somewhere, that you can add PBS into the wells with fixed cells and store at 4 degrees. But I was wondering if there is another way. And If you do remove the PBS for imaging, won't the fixed cells dry up quickly?
In addition, the fixed cells are to be imaged using a widefield, classical confocal, spinning disk confocal, and imagexpress.
If I understand your question, you are fixing cultured cells grown on plastic well plates, for later staining and imaging by microscopy? Presumably you are fixing with paraformaldehyde or something similar?
After fixing with PFA, you can switch to PBS and store at 4 degrees. They should be perfectly happy like this for several weeks until it is time to image them
Yeah.. but when imaging. do you still image with the pbs inside? or do i remove it?
Is your microscope inverted or upright? If it's inverted it shouldn't matter as you are viewing through the bottom of the dish. If it's upright you'll need to consider which magnification you want to use - some of the high powered objectives require oil immersion, which will interfere with your sample. At lower power it should be possible to view through the liquid (although the resolution won't be that great)
This helps greatly. I thank you alot for this.
I just started my phd and i am trying to do things differently.
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