Fig. 2

Characterization of 2PM and 3PM images through the deepest part of the adult D. dracula brain?the cerebellum-midbrain boundary

Excitation wavelengths of 920 nm at 2 MHz repetition rate and 1280 nm at 333 kHz repetition rate were used for 2PM and 3PM, respectively.

(A) Signal-to-background ratio comparison of horizontal and vertical blood vessels for 2PM and 3PM images at two depths inside the brain. In each line profile plot, the values are normalized to the maximum brightness of the horizontal blood vessel.

(B) Representative 3D reconstruction of 3PM (left) and 2PM (right) images of fluorescein-labeled blood vessels in the midbrain.

(C) Maximum projection of third harmonic generation (THG, left) and second harmonic generation (SHG, right) of all frames containing the skin. Signals were generated by 1280 nm excitation. Pigments produce very bright signal in the THG channel.

(D) Characterization of effective attenuation length inside the brain for 2PM and 3PM excitation wavelengths as described in the STAR Methods section.

(E) Axial (z) and lateral (xy) width of small blood vessel characterized in shallow (?100 ?m) and deep (?600 ?m) regions with 3PM.

(F) Axial (z) and lateral (xy) width of small blood vessel characterized in shallow (?100 ?m) and deep (?600 ?m) regions with 2PM.