Cellular Resolution Functional Imaging in Behaving Rats Using Voluntary Head Restraint

• Published in: Neuron (Cambridge, Mass.) Vol. 80; no. 2; pp. 371 - 384
• Main Authors: Scott, Benjamin B., Brody, Carlos D., Tank, David W.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.10.2013; Elsevier Limited
• Subjects: Animal cognition; Animals; Automation; Behavior; Brain; Conditioning, Operant - physiology; Evoked Potentials, Visual - physiology; Functional Neuroimaging - instrumentation; Functional Neuroimaging - methods; Head; Immobilization - psychology; Kinematics; Male; Molecular Imaging - methods; Neurons; Rats; Rodents; Visual Cortex - physiology
• ISSN: 0896-6273; 1097-4199
• DOI: 10.1016/j.neuron.2013.08.002

High-throughput operant conditioning systems for rodents provide efficient training on sophisticated behavioral tasks. Combining these systems with technologies for cellular resolution functional imaging would provide a powerful approach to study neural dynamics during behavior. Here we describe an integrated two-photon microscope and behavioral apparatus that allows cellular resolution functional imaging of cortical regions during epochs of voluntary head restraint. Rats were trained to initiate periods of restraint up to 8 s in duration, which provided the mechanical stability necessary for in vivo imaging while allowing free movement between behavioral trials. A mechanical registration system repositioned the head to within a few microns, allowing the same neuronal populations to be imaged on each trial. In proof-of-principle experiments, calcium-dependent fluorescence transients were recorded from GCaMP-labeled cortical neurons. In contrast to previous methods for head restraint, this system can be incorporated into high-throughput operant conditioning systems. •Rat voluntary head restraint trained by automated operant conditioning•Micron-scale brain repositioning during restraint using a kinematic mount•Two-photon cellular resolution calcium imaging from the same neurons on every trial•In vivo functional imaging of GECIs in awake, behaving rats Scott et al. describe a method combining high-throughput behavioral training with two-photon imaging. Using this technique, rats can be trained, in an automated fashion, to perform decision-making behaviors during voluntary head restraint, allowing neuronal dynamics to be imaged at cellular resolution.