Novel applications of fluorescence decay analysis in living tissue at single cell level

Fluorescence microscopy is a powerful tool to study the subcellular localisation and intracellular dynamics of fluorophore tagged proteins in living cells. Optical and spectroscopic technologies working at subcellular resolution with quantitative output are required for a deeper understanding of molecular processes in living cells and for the realization of predictive biology at a cellular and subcellular level. However, although established in the physical sciences, these techniques are rarely applied to cell biology especially in the plant sciences. This is due to several problems inherent in live-cell imaging, which will be specified in the first part of this volume. In the following, several approaches to circumvent these problems will be presented. This includes a novel method to suppress autofluorescence interference in fluorescence microscopy images of living cells. Additionally, the power of combining these approaches is demonstrated in several impressive examples.