Three-photon imaging of mouse brain structure and function through the intact skull

• Published in: Nature methods Vol. 15; no. 10; pp. 789 - 792
• Main Authors: Wang, Tianyu, Ouzounov, Dimitre G., Wu, Chunyan, Horton, Nicholas G., Zhang, Bin, Wu, Cheng-Hsun, Zhang, Yanping, Schnitzer, Mark J., Xu, Chris
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.10.2018; Nature Publishing Group
• Subjects: 631/1647/245/2225; 631/1647/245/2226; 631/1647/334/1874/345; 631/378; Animals; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Brain; Brain - anatomy & histology; Brain - physiology; Brief Communication; Calcium imaging; Cortex; Excitation; Female; Field of view; Frames per second; Functional anatomy; Image Processing, Computer-Assisted - methods; Imaging systems; Life Sciences; Male; Medical examination; Medical imaging; Methods; Mice; Mice, Inbred C57BL; Micrometers; Microscopy; Microscopy, Fluorescence, Multiphoton - methods; Neuroimaging; Neuroimaging - methods; Optical sectioning; Physiological aspects; Proteomics; Sectioning; Skull; Skull - anatomy & histology; Skull - physiology; Structure-function relationships; United States
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/s41592-018-0115-y

Optical imaging through the intact mouse skull is challenging because of skull-induced aberrations and scattering. We found that three-photon excitation provided improved optical sectioning compared with that obtained with two-photon excitation, even when we used the same excitation wavelength and imaging system. Here we demonstrate three-photon imaging of vasculature through the adult mouse skull at >500-μm depth, as well as GCaMP6s calcium imaging over weeks in cortical layers 2/3 and 4 in awake mice, with 8.5 frames per second and a field of view spanning hundreds of micrometers. Wang et al. demonstrate that the effects of aberrations and scattering caused by the mouse skull can be reduced with three-photon microscopy. Their approach allows structural and functional imaging of the brain through an intact skull.