In Vivo Superresolution Imaging of Neuronal Structure in the Mouse Brain.

ABSTRACT: OBJECTIVE:this study proposes and evaluates a technique for in vivo deep-tissue superresolution imaging in the light-scattering mouse brain at up to a 3.5 Hz 2-D imaging rate with a 21×21 ?m2 field of view. METHODS:we combine the deep-tissue penetration and high imaging speed of resonant laser scanning two-photon (2P) microscopy with the superresolution ability of patterned excitation microscopy. Using high-frequency intensity modulation of the scanned two-photon excitation beam, we generate patterned illumination at the imaging plane. Using the principles of structured illumination, the high-frequency components in the collected images are then used to reconstruct images with an approximate twofold increase in optical resolution. RESULTS:using our technique, resonant 2P superresolution patterned excitation reconstruction microscopy, we demonstrate our ability to investigate nanoscopic neuronal architecture in the cerebral cortex of the mouse brain at a depth of 120 ?m in vivo and 210 ?m ex vivo with a resolution of 119 nm. This technique optimizes the combination of speed and depth for improved in vivo imaging in the rodent neocortex. CONCLUSION:this study demonstrates a potentially useful technique for superresolution in vivo investigations in the rodent brain in deep tissue, creating a platform for investigating nanoscopic neuronal dynamics. SIGNIFICANCE:this technique optimizes the combination of speed and depth for improved superresolution in vivo imaging in the rodent neocortex.

SUBMITTER: Urban BE 

PROVIDER: S-EPMC5967880 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature