Deep Tissue Scattering Compensation with Three-Photon F-SHARP

Optical imaging techniques are widely used in biological research, but their penetration depth is limited by tissue scattering. Wavefront shaping techniques are able to overcome this problem in principle, but are often slow, and their performance depends on the sample. This greatly reduces their practicability for biological applications. Here we present a scattering compensation technique based on three-photon excitation, which converges faster than comparable two-photon (2P) techniques and works reliably even on densely labeled samples, where 2P approaches fail. To demonstrate its usability and advantages for biomedical imaging, we apply it to the imaging of dendritic spines on layer 5 neurons labeled with green fluorescent protein in an anesthetized mouse. (C) 2021 Optical Society of America under the terms of the OSA Open Access Publishing Agreement