Extended Data Fig. 6 |. AO improves in vivo 3P imaging of cortical neurons in the mouse brain.

a, Lateral and axial images of a neuronal cell body (Thy1-YFP-H), at 757 μm below dura, under 1300 nm excitation, without and with AO (same cell body as in Fig. 2a). Post-objective power: 17 mW. b, Signal profiles along the purple and yellow lines in a. c, Maximum intensity projection (MIP) of same neuron as in a, 747–767 μm below dura, under 1300 nm excitation, without and with AO. Post-objective power: 17 mW. d, MIP of the yellow square in c, at 747–757 μm below dura, without and with AO. Insets in d, zoomed-in views of dendrite in white box. 10× digital gain was applied to the inset without AO to improve visibility. Post-objective power: 20 mW. e, Lateral and axial images of a neuron in the mouse cortex (Thy1-GFP-M), at 687 μm below dura, under 1300 nm excitation, taken without and with AO. Post-objective power: 35 mW. f, Signal profiles along the purple and yellow lines in e. g, Corrective wavefront in e. h, MIP of a neuron in the mouse cortex (Thy1-GFP-M, different animal than in e), at 624–644 μm below dura, under 1300 nm excitation, without and with AO. Post-objective power: 13 mW. i, Signal profiles along the purple and blue lines in h. j, Corrective wavefront in h. Insets in a and e: spatial frequency space representation of the corresponding fluorescence images. Scale bars, 10 μm. Microscope objective: NA 1.05 25×. Representative results from 32 fields of view and 8 mice.