Computational and Optical Approaches to Multidimensional Live Cell Imaging Event

Abstract:

Light microscopy is providing key insights into cellular dynamics of normal and disease processes, a task being greatly facilitated by technical developments in optical probes and instrumentation. Fluorescent protein reporters allow virtually any protein to be labeled and thereby visualized in a cell, tissue or organism. Along with these developments in fluorescence probe technology, new optical techniques such as Fluorescence Resonance Energy Transfer (FRET), Multiphoton Laser-Scanning Microscopy (MPLSM), Second Harmonic Generation (SHG) imaging, Selective Plane Illumination Microscopy (SPIM) and Fluorescence Lifetime Imaging (FLIM), are revealing how individual cellular components are assembled into cytoplasmic machinery, and how this machinery functions. Our central aim is to develop hardware and software strategies for extracting all the data possible from the weak fluorescence signals that are typically obtained when observing living specimens. This involves the development and application of hardware and software for acquisition, analysis and visualization including feature extraction, exponential curve fitting, spectral unmixing and 3D visualization approaches. Much of this work is under the umbrella of our ImageJ2 software development to offer practical tools for developers and end user alike for the analysis of microscopy image data.