Extended Data Fig. 7: Evaluation of photobleaching lifetime of PF555 under PCA/PCD conditions in live COS7 cells.
From: Super-photostable organic dye for long-term live-cell single-protein imaging
a. Representative single-molecule images of HALO-EGFR labeled with CA-AF647, CA(O4)-PF555, and CA-TMR, both with and without the addition of PCA/PCD, on the PM of living COS7 cells. The images were acquired using a 561 nm laser with an intensity of 30 W/cm2 and an EM-CCD camera with a frame rate of 20 Hz. Photobleaching lifetime values were extracted from n = 7 cells across 7 independent culture batches. Scale bar = 10 µm. b. The graph depicts the time-dependent reduction in the number of fluorescence spots detected in each 50-ms frame for CA-AF647, CA(O4)-PF555, and CA-TMR on the PM of live COS7 cells. The black lines represent the best-fit single-exponential decay functions for each dye, and the derived exponential decay time constants are indicated. c. The photobleaching lifetime, represented as the exponential decay time constant, was calculated for CA-AF647, CA(O4)-PF555, and CA-TMR under both control and PCA/PCD-supplemented conditions. Photobleaching lifetime values were extracted from n = 7 cells across 7 independent culture batches. Data are presented as mean +/− SD.
