Actually, in my experiment, I was using 70% ethanol up till now as a fixing solvent for increasing the permeability of the nuclear membrane. Also, I have read about paraformaldehyde and some detergents (Triton-X100) for increasing the permeability so more accurate results are produced.
The cells I am using for my studies are cancer cell lines.
Any comparative studies using such system will be helpful.
Thank you
http://www.abcam.com/protocols/flow-cytometric-analysis-of-cell-cycle-with-propidium-iodide-dna-staining
Dear Joshi
You can use formaldehyde and glutaraldehyde for the increasement of permeability of nuclear membrane, you can also use organic solvent such as methanol and acetone for the same.
I assume you do PI staining for cell cycle analysis.
They work equally well but the choice depends on your experimental design. However, regardless which one you use, RNase must be used in combination with PI because PI stains both DNA and RNA. That is the way to ensure accurate results.
70% ethanol can fix and permeabilize cells at the same time. It saves time but doesn't allow surface staining for complicated flow cytometry analysis.
Paraformaldehyde is a fixative reagent only. So, the fixed cells must be permeabilized by Triton X100 or saponin. It allows surface staining.
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