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Synaptic plasticity, the ability for synapses to change their connection strength, is thought to underlie learning and memory. Cascades of biochemical reactions in dendritic spines, tiny (~0.1 femtoliter) postsynaptic compartments emanating from dendritic surface, trigger diverse forms of synaptic plasticity. These reactions are mediated by signaling networks consist of hundreds of species of signaling proteins. Our focus is to elucidate some of the operation principles of such signaling networks in dendritic spines using various optical techniques. First, we have been developing techniques to image activity of various proteins in single dendritic spines using 2-photon fluorescence lifetime imaging microscopy (2pFLIM) in combination with new biosensors extensively optimized for 2pFLIM. Using this technique, we have succeeded in imaging signaling proteins, CaMKII, HRas, RhoA and Cdc42, in single dendritic spines undergoing synaptic potentiation.