Fig. 1: E1GFP in vitro calibration. | Communications Biology

Fig. 1: E1GFP in vitro calibration.

From: Measuring pH in insulin secretory granules by phasor-based fluorescence lifetime imaging of a genetically encoded sensor

Fig. 1

a Lifetime decays of A′ (red) and A (blue) upon excitation at 405 nm and collection in the 480–550 nm range. Purified recombinant E1GFP was incubated in citrate/phosphate buffer at pH 4.85 or 8.0 to obtain almost pure A′ and A forms, respectively. b Representative FLIM images of recombinant E1GFP diluted in calibration buffer with the indicated pH. Images are pseudo-colored based on the lifetime calculated per pixel according to the color bar depicted in the figure. c Phasor plot analysis of the images of panel b. Phasor barycenters with SD and the corresponding linear regression fit are reported.

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