FluoQ: A Tool for Rapid Analysis of Multiparameter Fluorescence Imaging Data Applied to Oscillatory Events

The number of fluorescent sensors
and their use in living cells
has significantly increased in the past years. Yet, the analysis of
data from single cells or cell populations usually remains a very
time-consuming enterprise. Here, we introduce FluoQ, a new macro for
the image analysis software ImageJ, which enables fast analysis of
multiparameter time-lapse fluorescence microscopy data with minimal
manual input. FluoQ provides statistical analysis of all measured
parameters and delivers the results in multiple graphic and numeric
displays. We demonstrate the power of FluoQ by applying the macro
to data analysis in the development and optimization of novel FRET
reporters for monitoring the performance of calcium/calmodulin-binding
inositol trisphosphate kinases A and B (ITPKA and ITPKB) in HeLa cells.
We find that conformational changes in the ITPKA-based sensor follow
receptor-mediated calcium oscillations. This indicates that ITPKA
contributes to the regulation of intracellular calcium transients
by limiting inositol trisphosphate levels.