A discussion of wavelet-based analysis of FRET (Forster resonance energy transfer) microscopy describes a technique that enhances quantitative analysis of the spatiotemporal dynamics of cellular microdomains. FRET is very useful for imaging when assessing nanometer-scale associations within live cells, FRET entails the monitoring of the fluorescence of an acceptor fluorophore when energy is transferred from an excited fluorophore that is in proximity (within 1 to 10nm). FRET microscopy is also possibly useful for detection or protein-protein interactions and as conformational changes in engineered reporter constructs. Even with enhanced FRET-based imaging technology, quantitative characterization and analysis of the spatio-temporal dynamics of signaling microdomains is still challenging. Researchers have proposed a wavelet transform-based approach for detection and quantification of changes in functional cellular microdomains. An optimal mathematical transform can remap a signal to extract new information about the properties of the initial signal. Multi-resolution properties of the wavelet transform make it especially suitable for signals that contain transient properties. The goal was to exploit this property of the wavelet transform for analysis of FRET microscopic images of COA1 cells expressing a membrane-localized genetically encoded reporter of protein kinase C activity.