We describe an adaptive optics method that modulates the intensity or phase of light rays at multiple pupil segments in parallel to determine the sample-induced aberration. Applicable to fluorescent protein-labeled structures of arbitrary complexity, it allowed us to obtain diffraction-limited resolution in various samples in vivo. For the strongly scattering mouse brain, a single aberration correction improved structural and functional imaging of fine neuronal processes over a large imaging volume.
Bibliographical noteFunding Information:
We thank our colleagues at Janelia Farm Research Campus, Howard Hughes Medical Institute: E. Betzig for helpful discussions; H. Dana, B. MacLennan, G. Ranganathan, K. Smith for help with mice samples; and P. Keller and M. Ahrens for providing zebrafish samples. We thank C. Fang-Yen for advice on C. elegans samples. This work was supported by Howard Hughes Medical Institute.