Characterization and adaptive optical correction of aberrations during in vivo imaging in the mouse cortex

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 109; no. 1; pp. 22 - 27
• Main Authors: Ji, Na, Sato, Takashi R, Betzig, Eric
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 03.01.2012; National Acad Sciences
• Subjects: Adaptive optics; Animals; Biological Sciences; Brain; Calcium; Calcium - metabolism; Cerebral Cortex - anatomy & histology; cortex; Fluorescence; image analysis; Imaging; Imaging, Three-Dimensional - methods; Medical imaging; Mice; Microscopy; Neuroimaging; Neurons; Optics; Optics and Photonics; Physical Sciences; Rodents; Skull; Wave excitation
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1109202108

The signal and resolution during in vivo imaging of the mouse brain is limited by sample-induced optical aberrations. We find that, although the optical aberrations can vary across the sample and increase in magnitude with depth, they remain stable for hours. As a result, two-photon adaptive optics can recover diffraction-limited performance to depths of 450 μm and improve imaging quality over fields of view of hundreds of microns. Adaptive optical correction yielded fivefold signal enhancement for small neuronal structures and a threefold increase in axial resolution. The corrections allowed us to detect smaller neuronal structures at greater contrast and also improve the signal-to-noise ratio during functional Ca2+ imaging in single neurons.