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Single-molecule imaging of S65T-GFP molecules by using confocal excitation. Images are 10 × 10 μm2, 300 × 300 pixels per image, and acquisition time of 1.6 ms per pixel. Spot size is ≈300 nm. (a) Excitation intensity is 1.2 kW/cm2. Most molecules emit in a rather stable fashion, with little blinking during the scanning. (b) Excitation intensity is 7 kW/cm2. Molecular emission displays increased blinking. The majority of the molecules stop emission after a few lines of scanning.
