I prepared my samples with ethanol, RNase and PI. However I could not use the Flow Cytometry machine because of an technical problem. I covered my samples with foil and put them in +4. As my cells are fixated I don't question them but I'm not sure about the stability of PI. I don't know if PI is stable for longer times.
If you protect samples from light and keep them at 4C I would say you have good chances to get your data within a week.
Hello, Zeynep!
Unfixed cell preparations, whether derived from suspension or monolayer cultures or from solid tumors, are stable for 2 or more weeks if stored at 4 degrees C between flow cytometric analyses and histograms are usually only minimally altered if the stained cell samples are stored for 1-2 months at 4 degrees C.
Article A stable propidium iodide staining procedure for flow cytometry
Dear @zeynep
Theoretically, you may keep them for 1-2 weeks, but as per my experience, we add PI just before the acquisition (that gives best results). but keeping even fixed cells more then 2-3 days would impact cells. PI is very sticky dye, you will find much positivity in PI chennal.
Try to acquire the sample as soon
In my experience, storage of EtOH-fixed cells before analysis worked up to 2 weeks. However, I would recommend to stain with PI only immediately before flow cytometric analysis, which will give you the best results. Intercalation might have an impact on DNA stability when stored for a longer period of time.
Since PI is a viability stain I wouldn’t stain after fixing.. stain with PI before fixing or use a fixable live/dead stain. Perform the FACs within a week of preparation for ideal results
Oftenly the problem in flow cytometry is because of the lower cell density/concentration as cell cycle analysis is performed in growing cells, please check it also.
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