Extended Data Fig. 3: Photostability of RFP variants.
From: mScarlet3: a brilliant and fast-maturing red fluorescent protein
a: Wide-field bleaching of mScarlet variants without time-axis correction, b: same data as a) but y-axis scaled according to cellular brightness; c: confocal bleaching of mScarlet variants; d: confocal bleaching of mScarlet variants and other RFPs. In order to fairly compare the photostability of different RFP variants under confocal imaging conditions, the time axis is corrected in c and d for spectral properties relative to mScarlet to mimic an identical overall emission rate (photons/s) for all RFPs at the start of the experiment. e: Blue light-induced photochromicity of RFP variants. The RFP variants were expressed in HeLa cells and continuously exposed with 550 nm excitation light. At t = 15, 31, 46 and 52 s, the 550 nm excitation light interrupted by a brief excitation at 470 nm for 1 s (not recorded on time axis) after which the 550 nm excitation was continued. The sudden increase after blue light excitation is caused by photochromic behavior.
