Optimization of Fluorescent Proteins

Nowadays, fluorescent protein (FP) variants have been engineered to fluoresce in all different colors; to display photoswitchable, or photochromic, behavior; or to show yet other beneficial properties that enable or enhance a still growing set of new fluorescence spectroscopy and microcopy techniques. This has allowed the (in situ) study of biomolecules with unprecedented resolution, specificity, sensitivity, and ease of labeling. However, brighter FPs, more photostable FPs, and FPs that display an even better compatibility with biophysical microspectroscopic techniques are still highly desired. The key characteristics of FPs—absorption spectrum, emission spectrum, brightness, fluorescence lifetime, maturation rate, oligomeric state, photostability, pH sensitivity, and functionality in protein fusions—determine their application. This chapter will describe these key features and present several experimental protocols to optimize them.

Source: Springer protocols feed by Imaging/Radiology - October 14, 2013 Category: Radiology Source Type: news