The phasor calculator (right bottom panel) shows the mTQ2 and mTFP1 S-G coordinates and a black circle designating the point where the VS sits. c The average lifetime approach is shown pixel-by-pixel for the same example as in (a). The dotted square shows the VS and the corresponding...
All pixels with values < 0.1% in “a1[%]” and “a2[%]” images were converted to not-a-number (NaN) (Image>Adjust>Threshold…). 6. ‘Pixel Intensity’ image was multiplied by “a1[%]” or “a2[%]” images (Process>Image Calculator…) to generate two images that contained ...
We converted the phase and modulation lifetime values of all calibration experiments to polar coordinates34, which resulted in a straight line on a polar (or phasor) plot (Supplementary Fig. S10a). For each calcium concentration, we calculated the fraction of the sensor in the high lifetime ...