The excitation wavelength of Fura-2 is overlapped with GFP....

Apr 25, 2012
protocol Protocol: Calcium Imaging
The excitation wavelength of Fura-2 is overlapped with GFP. If the cells to be measured express GFP, does this affect the measurement? what is the difference between Fura-2(Calcium free) and Fura-2(Calcium bound)?
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Mingye Feng Author Answered Apr 25, 2012

The Johns Hopkins University School of Medicine

GFP should have minor effects on Fura-2 measurement, because its major excitation peak is over 450nm, while Fura-2 is excited at 340 and 380nm. But a GFP-only control would be necessary for the measurement.

Fura-2(Calcium free) and Fura-2(Calcium bound) have different excitation wavelength. When Fura-2 is bound to calcium, its excitation peak is 340nm. When Fura-2 is not bound to calcium, it will be 380nm.
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