Optical Multilayer Interference Tomography Compatible with Tape-Based Serial SEM for Mesoscale Neuroanatomy

Understanding the complexity of animals’ behaviors and intellect relies on decoding the underlying structure and wiring diagram of their brains. Various imaging techniques have been developed to tackle this challenge, yet they still have limitations in mapping all cells in a brain region at mesoscale level. Here we propose an optical multilayer interference tomography (OMLIT) method with serial sectioning capability for mesoscale neuroanatomy. With OMLIT, we can distinguish and reconstruct all cell bodies, large branches of dendrites and myelinated axons, and the vasculature in the region of interest, enabling fast mapping of cell density and classification of certain cell types. Furthermore, using the same form of ultrathin sample section library, our method offers seamless compatibility with tape-based serial scanning electron microscopy, providing a correlative approach to reconstruct both mesoscale and microscale brain atlases in the same tissue.