Serial Two-Photon Tomography Imaging of the whole marmoset brain for neuroanatomical analyses

Abstract

Serial Two photon tomography (STPT) imaging is a technique to image a mass of tissue in its three-dimensional shape by combining two-photon imaging with automatic stage control and microtome slicing. We successfully implemented this technique for tracing the axonal projections in the marmoset brain. Here we describe the detailed experimental procedures that realized reliable volumetric imaging of the whole marmoset brain. We also introduce auxiliary histological techniques including visualization of myelin structure by simple light reflection, and immunological detection of non-fluorescent anterograde and retrograde tracers, which further enhance the utility of STPT data.