JournalNeural Excitability, Synapses, and Glia

Automated Remote Focusing, Drift Correction, and Photostimulation to Evaluate Structural Plasticity in Dendritic Spines

A: Individual imaging locations are identified on a single cell. B: Parallel stimulation of dendritic spines results in changes in spine volume. C: Optimal locations for photostimulation are automatically identified on each spine prior to stimulation.

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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.


Smirnov, M.S., Evans, P.R., Garrett, T.R., Yan, L., and Yasuda, R. (2017). Automated Remote Focusing, Drift Correction, and Photostimulation to Evaluate Structural Plasticity in Dendritic Spines. PLOS ONE 12, e0170586.
DOI: http://dx.doi.org/10.1371%2Fjournal.pone.0170586

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