Smirnov Michael S, Evans Paul R, Garrett Tavita R, Yan Long, Yasuda Ryohei
Neuronal Signal Transduction, Max Planck Florida Institute for Neuroscience, Jupiter, Florida, United States of America.
Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia, United States of America.
PLoS One. 2017 Jan 23;12(1):e0170586. doi: 10.1371/journal.pone.0170586. eCollection 2017.
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.
树突棘的长期结构可塑性在突触可塑性中起关键作用，而突触可塑性是学习和记忆的细胞基础。生化步骤由棘突中复杂的信号蛋白网络介导。双光子成像技术与双光子谷氨酸解笼相结合，使研究人员能够在单个树突棘中诱导和量化结构可塑性。然而，这种方法费力且缓慢，不适用于对结构可塑性所需因子进行高通量筛选。在此，我们介绍一个基于MATLAB构建的模块，该模块用于Scanimage，可自动跟踪、成像和刺激多个树突棘。我们结合电可调谐透镜和漂移校正算法，以快速连续地跟踪目标棘突并校正样品移动。通过简单的用户界面设计定制的多位点实验，我们能够使用谷氨酸解笼对多个树突棘进行充分成像并产生目标可塑性。我们的方法成本低廉、开源，并且在量化树突棘结构可塑性的通量方面提高了多达五倍。
