I would like to put PI on tissue slices and then fix them to perform IF. As a second readout for cell death, I will also use cleaved caspase 3 staining. I am not sure if PFA maintains PI in tissue/cells.
Hello Alexandre,
I use NBF rather than PFA and the PI signal is significantly lost after fixation.
Propidium iodide (PI) is a commonly used fluorescent dye that intercalates with DNA and is often used to assess cell viability and cell death. Fixation with paraformaldehyde (PFA) is a standard method for preserving cellular structures and enabling immunofluorescence (IF) staining. However, there are considerations to keep in mind when using PI and PFA together.
While PFA fixation can preserve overall tissue morphology and certain antigens, it can affect the fluorescence signal of PI. PFA fixation may cause a decrease in PI fluorescence intensity, potentially resulting in an underestimation of cell death. Additionally, PFA can affect the accessibility of PI to intracellular DNA, potentially reducing its staining efficiency.
To mitigate these issues, there are a few strategies you can consider:
Hi Alexandre,
Yes you can, we used it in our lab and got a good signal of PI in a cell line. Make sure to add antifade.
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