Extended Data Fig. 6: Photoactivation and spontaneous recovery of CA(O4)-PF555.
From: Super-photostable organic dye for long-term live-cell single-protein imaging
a, d. TIRF images of Halo-EGFR in live COS7 cells labeled with CA(O4)-PF555 after 30 minutes of photobleaching with snapshots taken every 10 s over 25 minutes without (c) or with (d) a 10 s 405 nm laser pulse between snapshots. Scale bar = 20 μm. b, e. Intensity profiles from acquired images were extracted and fitted with an exponential decay function to quantify the rate and amount of spontaneous recovery (e) and photoactivation (f). c, f. Photobleaching lifetime of spontaneous recovered species (c) and photoactivated species (f) calculated by exponential decay fitting. g. The rate (left panel), amount (middle panel) and photobleaching lifetime (right panel) of photoactivation (blue) and spontaneous recovery (red) were determined by fitting the exponential decay function to the intensity profile of the CA(O4)-PF555 signal. The rate, amount and photobleaching lifetime measurements were repeated on n = 10 cells across 3 independent culture batches. Data are presented as mean +/− SD. P values are the result of a two-sided t-test. Left panel: ****P = 4.4 × 10−16. Middle panel: P = 0.0791 (ns: not significant). Right panel: ****P = 6.2 × 10−14. h. Intensity profile of CA(O4)-PF555 (black line) under repeated photoactivation by 10 s 405 nm laser pulses (pink line).
