PT  - JOURNAL ARTICLE
AU  - Michael S Smirnov
AU  - Paul R Evans
AU  - Tavita R Garrett
AU  - Long Yan
AU  - Ryohei Yasuda
TI  - Automated remote focusing, drift correction, and photostimulation to evaluate structural plasticity in dendritic spines
AID  - 10.1101/083006
DP  - 2016 Jan 01
TA  - bioRxiv
PG  - 083006
4099  - http://biorxiv.org/content/early/2016/10/24/083006.short
4100  - http://biorxiv.org/content/early/2016/10/24/083006.full
AB  - 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.