RT Journal Article
SR Electronic
T1 Recording neural activity in unrestrained animals with 3D tracking two photon microscopy
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 213942
DO 10.1101/213942
A1 Doycho Karagyozov
A1 Mirna Mihovilovic Skanata
A1 Amanda Lesar
A1 Marc Gershow
YR 2017
UL http://biorxiv.org/content/early/2017/11/19/213942.abstract
AB Optical recordings of neural activity in behaving animals can reveal the neural correlates of decision making, but such recordings are compromised by brain motion that often accompanies behavior. Two-photon point scanning microscopy is especially sensitive to motion artifacts, and to date, two-photon recording of activity has required rigid mechanical coupling between the brain and microscope. To overcome these difficulties, we developed a two-photon tracking microscope with extremely low latency (360 μs) feedback implemented in hardware. We maintained continuous focus on neurons moving with velocities of 3 mm/s and accelerations of 1 m/s2 both in-plane and axially, allowing high-bandwidth measurements with modest excitation power. We recorded from motor- and inter-neurons in unrestrained freely behaving fruit fly larvae, correlating neural activity with stimulus presentation and behavioral outputs. Our technique can be extended to stabilize recordings in a variety of moving substrates.