Cellular resolution functional imaging in behaving rats using voluntary head restraint.
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.
Princeton Neuroscience Institute, Princeton University, Princeton, NJ 08544, USA; Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA; Bezos Center for Neural Circuit Dynamics, Princeton University, Princeton, NJ 08544, USA.
Benjamin B
BB
Scott
Carlos D
CD
Brody
David W
DW
Tank
D000818:Animals; D003216:Conditioning, Operant / Q000502:physiology*; D005074:Evoked Potentials, Visual / Q000502:physiology; D059907:Functional Neuroimaging / Q000295:instrumentation / Q000379:methods*; D006257:Head; D007103:Immobilization / Q000523:psychology*; D008297:Male; D057054:Molecular Imaging / Q000379:methods; D051381:Rats; D014793:Visual Cortex / Q000502:physiology