Hello everyone.
I have performed cell cycle analysis by FCM using FxCycle PI/RNase staining solution from Invitrogen for MSCs. I fixed my cells with 70% ethanol for 2 hours at 4 degree C before staining with the solution.
My results showed 2 peaks which commonly known as G0/G1 peak and also G2/M peak, however the G2/M peak was very far from the G0/G1 peak and not showing 4N DNA content (attached file).
What would be the probable causes?
I am thankful for any tips and advice.
The cause of the discrepancy in your cell cycle analysis results could be due to several factors, including improper cell fixing, staining or sample preparation, or issues with instrumentation or data analysis. In particular, the gating strategy can greatly impact the accuracy of cell cycle analysis results, so it is possible that incorrect gating could be a contributing factor. I would recommend checking your gating strategy and reviewing your sample preparation and staining protocols to ensure that they were performed correctly. Additionally, it might be helpful to consult with a specialist or perform a quality control check with a known cell line to ensure that the results are accurate.
Maria Arshad Thank you so much for the advice. I will re-check my gating strategy again and reconsider other factors.
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