Hi everyone,
I used propidium iodide as nuclear staining With concentration (5 µg/ml) for DRG nonneuronal cells for 2 hours pre-fixation to detect dead cells. I also used DAPI for staining all cells nuclei. I have consistent issue with all slides. I found that PI gave red fluorescence signals for some dead cells in one field but other cells nuclei with symptoms of cell death such as nuclear fragmentation or condensed nuclei did not have any positive PI signals ( white arrows in the micrographs). Therefore, I am wondering why Pi did not give signals for cells with nuclear fragmentation as shown here in these attached micrographs. I used PI for the first time and I do not have experience with PI staining. Could you please somebody used PI or have experience with PI staining interpret why there is no PI signals with cells have fragmented nuclei or pycnotic nuclei? or maybe the problem in PI staining itself or the concentration. Thanks in advance
Amira Elfarnawany
Propidium iodide only stains dead cells. Your question relates to the theory of hydration of Propidium iodide and DNA in dead and living cells. Living cells (open systems) of our body live when the energy of quantum and thermal fluctuations of water are approximately equal. Under these conditions, Propidium iodide fluorescence excitation does not work. In dead cells (closed systems), the equality of these energies is violated and the cells become stained. You can read about this theory in our works.
Preprint Nuclear quantum effect in aqueous micellar surfactant solutions
Thesis Mirgorod Yuri. QUANTUM NUCLEAR EFFECT IN AQUEOUS IONIC SURFA...
Yuri Mirgorod
Dear. prof. Mirgorod,
Thanks so much for your answer. everything is clear now.
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