@article {45787, title = {Anesthetized- and awake-patched whole-cell recordings in freely moving rats using UV-cured collar-based electrode stabilization.}, journal = {Nature Protocols}, volume = {9}, year = {2014}, month = {2014 Nov 06}, pages = {2784-95}, abstract = {

Intracellular recording allows precise measurement and manipulation of individual neurons, but it requires stable mechanical contact between the electrode and the cell membrane, and thus it has remained challenging to perform in behaving animals. Whole-cell recordings in freely moving animals can be obtained by rigidly fixing ({\textquoteright}anchoring{\textquoteright}) the pipette electrode to the head; however, previous anchoring procedures were slow and often caused substantial pipette movement, resulting in loss of the recording or of recording quality. We describe a UV-transparent collar and UV-cured adhesive technique that rapidly (within 15 s) anchors pipettes in place with virtually no movement, thus substantially improving the reliability, yield and quality of freely moving whole-cell recordings. Recordings are first obtained from anesthetized or awake head-fixed rats. UV light cures the thin adhesive layers linking pipette to collar to head. Then, the animals are rapidly and smoothly released for recording during unrestrained behavior. The anesthetized-patched version can be completed in \~{}4-7 h (excluding histology) and the awake-patched version requires \~{}1-4 h per day for \~{}2 weeks. These advances should greatly facilitate studies of neuronal integration and plasticity in identified cells during natural behaviors.

}, issn = {1750-2799}, doi = {10.1038/nprot.2014.190}, author = {Lee, Doyun and Shtengel, Gleb and Osborne, Jason E and Lee, Albert K} } @article {6885, title = {RIVETS: A mechanical system for in vivo and in vitro electrophysiology and imaging.}, journal = {PLoS One}, volume = {9}, year = {2014}, month = {2014 Feb 14}, pages = {e89007}, abstract = {

A number of recent studies have provided compelling demonstrations that both mice and rats can be trained to perform a variety of behavioral tasks while restrained by mechanical elements mounted to the skull. The independent development of this technique by a number of laboratories has led to diverse solutions. We found that these solutions often used expensive materials and impeded future development and modification in the absence of engineering support. In order to address these issues, here we report on the development of a flexible single hardware design for electrophysiology and imaging both in brain tissue in vitro. Our hardware facilitates the rapid conversion of a single preparation between physiology and imaging system and the conversion of a given system between preparations. In addition, our use of rapid prototyping machines ("3D printers") allows for the deployment of new designs within a day. Here, we present specifications for design and manufacturing as well as some data from our lab demonstrating the suitability of the design for physiology in behaving animals and imaging in vitro and in vivo.

}, issn = {1932-6203}, doi = {10.1371/journal.pone.0089007}, author = {Osborne, Jason E and Dudman, Joshua T} } @article {1645, title = {Two-photon calcium imaging from head-fixed Drosophila during optomotor walking behavior.}, journal = {Nature Methods}, volume = {7}, year = {2010}, month = {2010 Jul}, pages = {535-40}, abstract = {

Drosophila melanogaster is a model organism rich in genetic tools to manipulate and identify neural circuits involved in specific behaviors. Here we present a technique for two-photon calcium imaging in the central brain of head-fixed Drosophila walking on an air-supported ball. The ball{\textquoteright}s motion is tracked at high resolution and can be treated as a proxy for the fly{\textquoteright}s own movements. We used the genetically encoded calcium sensor, GCaMP3.0, to record from important elements of the motion-processing pathway, the horizontal-system lobula plate tangential cells (LPTCs) in the fly optic lobe. We presented motion stimuli to the tethered fly and found that calcium transients in horizontal-system neurons correlated with robust optomotor behavior during walking. Our technique allows both behavior and physiology in identified neurons to be monitored in a genetic model organism with an extensive repertoire of walking behaviors.

}, keywords = {Animals, Brain, Calcium, Computer-Assisted, Drosophila melanogaster, Fluorescence, Green Fluorescent Proteins, Image Processing, Motion, Motor Activity, Neurons, Signal Transduction, Walking}, issn = {1548-7105}, doi = {10.1038/nmeth.1468}, author = {Seelig, Johannes D and Chiappe, M Eugenia and Lott, Gus K and Dutta, Anirban and Osborne, Jason E and Reiser, Michael B and Jayaraman, Vivek} }