Multi-neuronal recording in unrestrained animals with all acousto-optic random-access line-scanning two-photon microscopy

Frontiers in Neuroscience

Neuroscience

Doycho Karagyozov¹  Rui Wu¹  Paul McNulty¹  Mirna Mihovilovic Skanata¹  Akihiro Yamaguchi¹  Marc Gershow² 

[1] Department of Physics, New York University, New York, NY, United States;Department of Physics, New York University, New York, NY, United States;Center for Neural Science, New York University, New York, NY, United States;Neuroscience Institute, New York University, New York, NY, United States;

关键词: two-photon;    drosophila;    tracking microscopy;    TAG lens;    calcium imaging;    acousto-optic;    drosophila larva;    motor system;   

DOI  :  10.3389/fnins.2023.1135457

 received in 2022-12-31, accepted in 2023-05-18,  发布年份 2023

来源: Frontiers

【 摘 要 】

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.

【 授权许可】

Unknown   
Copyright © 2023 Yamaguchi, Wu, McNulty, Karagyozov, Mihovilovic Skanata and Gershow.