Serial two-photon tomography for automated ex vivo mouse brain imaging

Abstract

Automated tissue sectioning and two-photon imaging of fluorescently labeled and fixed mouse brains allows high-resolution tomographic imaging of the entire brain. The authors demonstrate performance using multiple GFP mouse lines, dye-based retrograde tracing and viral anterograde tracing. Here we describe an automated method, named serial two-photon (STP) tomography, that achieves high-throughput fluorescence imaging of mouse brains by integrating two-photon microscopy and tissue sectioning. STP tomography generates high-resolution datasets that are free of distortions and can be readily warped in three dimensions, for example, for comparing multiple anatomical tracings. This method opens the door to routine systematic studies of neuroanatomy in mouse models of human brain disorders.