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Automated remote focusing, drift correction, and photostimulation to evaluate structural plasticity in dendritic spines

Michael S Smirnov, Paul R Evans, Tavita R Garrett, Long Yan, Ryohei Yasuda
doi: https://doi.org/10.1101/083006
Michael S Smirnov
1Neuronal Signal Transduction, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America
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  • For correspondence: Michael.Smirnov@mpfi.org
Paul R Evans
2Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia, United States of America
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Tavita R Garrett
1Neuronal Signal Transduction, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America
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Long Yan
3Light Microscopy Core, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America
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Ryohei Yasuda
1Neuronal Signal Transduction, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America
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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.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted October 24, 2016.
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Automated remote focusing, drift correction, and photostimulation to evaluate structural plasticity in dendritic spines
Michael S Smirnov, Paul R Evans, Tavita R Garrett, Long Yan, Ryohei Yasuda
bioRxiv 083006; doi: https://doi.org/10.1101/083006
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Automated remote focusing, drift correction, and photostimulation to evaluate structural plasticity in dendritic spines
Michael S Smirnov, Paul R Evans, Tavita R Garrett, Long Yan, Ryohei Yasuda
bioRxiv 083006; doi: https://doi.org/10.1101/083006

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