Scott Tighe wrote- >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. The emission wavelength of the J-aggregates can't shift, at least not enough to go from red to green. What you are seeing over time is fewer aggregates in the cells in your sample, probably because dye is washing out and/or because the mitochondria are depolarizing. -Howard
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