Dear Flowers: Interesting observation we have noticed. When using JC1 as a mitochondial membrane potential indicator, its emission wavelength of its J-aggregates is dramatically influenced by temperature. Cells which contain J-aggregates(orange- high potential) when analyzed at 4 C will maintain orange, however if we allow them to remain on the flow for a period of time for the cells/tubes to increase in temp toward room temp, the color shifts to green. Has anyone also experienced this? Our suspension buffer is Hanks and PBS... it does seem to matter. Cells: C10 mouse lung epi Cultural:CMRL 37C CO2 5% Trypsinized>neutralized in media>spin> resuspend in JC1@10uM. Also tried a PBS wash step. No help. Thanks Scott Tighe Vermont Cancer Center
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