Figure 2.

Empirically determined optimal bleaching conditions for one-photon (1PE) and two-photon (2PE) excitations. For each condition, both the xy-plane and the orthogonal view of the zy-plane are shown. Additionally, normalized intensity profiles registered along the z-direction (white solid lines) are included. Upper panel: 1PE with a laser power of 8.7 μW, a pixel dwell time of 2.09 μs, and a repetition of (A) 3, (B) 10, and (C) 20 frames. Lower panel: 2PE with (D) a laser power of 3.9 mW, a pixel dwell time of 2.09 μs, and a repetition of seven frames and (E) a laser power of 6.3 mW, a pixel dwell time of 2.09 μs, and a repetition of five frames or (F) a repetition of four frames. Pixel sizes varied depending on the area selected. All laser powers were measured at the objective. Scale bars: 5 μm.