Random-Access Two-Photon Microscopy Dataset: Neural Recording in Moving Larval Drosophila

This repository contains datasets and MATLAB scripts used for generating figures in the paper titled 'Multi-neuronal recording in unrestrained animals with acousto-optic two-photon microscopy,' published in Frontiers of Neuroscience in 2023. These datasets were also used in Akihiro Yamaguchi's dissertation, "Two-Photon Microscopy for Recording Multi-Neuronal Activity in Behaving, Untethered, Intact Animals."
To understand how neural activity encodes and coordinates behavior, it is desirable to record multi-neuronal activity in freely behaving animals. Imaging in unrestrained animals is challenging, especially for those, like larval Drosophila melanogaster, whose brains are deformed by body motion. A previously demonstrated two-photon tracking microscope recorded from individual neurons in freely crawling Drosophila larvae but faced limits in multi-neuronal recording. Here we demonstrate a new tracking microscope using acousto-optic deflectors (AODs) and an acoustic GRIN lens (TAG lens) to achieve axially resonant 2D random access scanning, sampling along arbitrarily located axial lines at a line rate of 70 kHz. With a tracking latency of 0.1 ms, this microscope recorded activities of various neurons in moving larval Drosophila CNS and VNC including premotor neurons, bilateral visual interneurons, and descending command neurons. This technique can be applied to the existing two-photon microscope to allow for fast 3D tracking and scanning.