title: Automated Remote Focusing, Drift Correction, and Photostimulation to Evaluate Structural Plasticity in Dendritic Spines.
abstract: Long-term structural plasticity of dendritic spines plays a key role in synaptic plasticity, the cellular basis for learning and memory. The biochemical step is mediated by a complex network of signaling proteins in spines. Two-photon imaging techniques combined with two-photon glutamate uncaging allows researchers to induce and quantify structural plasticity in single dendritic spines. However, this method is laborious and slow, making it unsuitable for high throughput screening of factors necessary for structural plasticity. Here we introduce a MATLAB-based module built for Scanimage to automatically track, image, and stimulate multiple dendritic spines. We implemented an electrically tunable lens in combination with a drift correction algorithm to rapidly and continuously track targeted spines and correct sample movements. With a straightforward user interface to design custom multi-position experiments, we were able to adequately image and produce targeted plasticity in multiple dendritic spines using glutamate uncaging. Our methods are inexpensive, open source, and provides up to a five-fold increase in throughput for quantifying structural plasticity of dendritic spines.
authors.affiliation: Neuronal Signal Transduction, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America.
authors.forename: Michael S
authors.identifier: http://orcid.org/0000-0003-2248-8863
authors.initials: MS
authors.lastname: Smirnov
authors.affiliation: Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia, United States of America.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Evans
authors.affiliation: Neuronal Signal Transduction, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America.
authors.forename: Tavita R
authors.initials: TR
authors.lastname: Garrett
authors.affiliation: Light Microscopy Core, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America.
authors.forename: Long
authors.initials: L
authors.lastname: Yan
authors.affiliation: Neuronal Signal Transduction, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America.
authors.forename: Ryohei
authors.initials: R
authors.lastname: Yasuda
mesh_terms: D000818:Animals; D001331:Automation; D049229:Dendritic Spines / Q000502:physiology / Q000528:radiation effects*; D008027:Light; D051379:Mice; D008810:Mice, Inbred C57BL; D009473:Neuronal Plasticity / Q000502:physiology*