I am having quite a bit of difficulty using this probe. It seems that there is no standardized way to analyze the data in Flowjo. I have also read many papers that say you need to compensate if you use the 488 nm laser, but how do you compensate with just one dye? Additionally, how do you determine the ratio of red/green fluorescence? Is it simply the MFI red/MFI green or does the calculation require normalization to the positive control?
Dear Sir. Concerning your issue about the performance of JC-1 analysis. I think the following below links may help you in your analysis:
https://www.caymanchem.com/pdfs/10009172.pdf
https://www.biotechniques.com/multimedia/archive/00074/Accuri-FP-Flow_Cytom_74531a.pdf http://www.abcam.com/ps/products/113/ab113850/documents/ab113850_7%20Nov%2013_JC%20(website).pdf
Thanks
Hi Isam,
Thank you for the response. My issues do not concern the staining and processing with a flow cytometer, but instead the analysis in Flow Jo. Many papers I have read indicate that they use a red:green ratio for quantitative analysis, but do not explain how they determined this ratio. This is where my confusion is coming from.
Hi Destini,
May I ask if you solved the problem now? I have the same confusion as how to calculate the red/green ratio. I searched a lot but got nothing useful.
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