High-Speed Neural Imaging with Synaptic Resolution: Bessel Focus Scanning Two-Photon Microscopy and Optical-Sectioning Widefield Microscopy

AbstractBrain is composed of complex networks of neurons that work in concert to underlie the animal’s cognition and behavior. Neurons communicate via structures called synapses, which typically require submicron spatial resolution to visualize. To understand the computation of individual neurons as well as neural networks, methods that can monitor neuronal morphology and function in vivo at synaptic spatial resolution and sub-second temporal resolution are required. In this chapter, we discuss the principles and applications of two enabling optical microscopy methods: two-photon fluorescence microscopy equipped with Bessel focus scanning technology and widefield fluorescence microscopy with optical sectioning ability, both of which could be combined with optogenetic stimulation for all optical interrogation of neural circuits. Details on their design and implementation, as well as example applications, are presented.