Open AccessGenetically encodable fluorescent protein markers in advanced optical imaging
Author(s) -
Karin Nienhaus,
G. Ulrich Nienhaus
Publication year - 2022
Publication title -
methods and applications in fluorescence
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 25
ISSN - 2050-6120
DOI - 10.1088/2050-6120/ac7d3f
Subject(s) - fluorescent protein , green fluorescent protein , toolbox , computational biology , fluorescence microscope , fluorescence , optical imaging , protein engineering , fluorescence lifetime imaging microscopy , nanotechnology , microscopy , biology , computer science , gene , genetics , biochemistry , materials science , physics , optics , enzyme , programming language
Optical fluorescence microscopy plays a pivotal role in the exploration of biological structure and dynamics, especially on live specimens. Progress in the field relies, on the one hand, on technical advances in imaging and data processing and, on the other hand, on progress in fluorescent marker technologies. Among these, genetically encodable fluorescent proteins (FPs) are invaluable tools, as they allow facile labeling of live cells, tissues or organisms, as these produce the FP markers all by themselves after introduction of a suitable gene. Here we cover FP markers from the GFP family of proteins as well as tetrapyrrole-binding proteins, which further complement the FP toolbox in important ways. A broad range of FP variants have been endowed, by using protein engineering, with photophysical properties that are essential for specific fluorescence microscopy techniques, notably those offering nanoscale image resolution. We briefly introduce various advanced imaging methods and show how they utilize the distinct properties of the FP markers in exciting imaging applications, with the aim to guide researchers toward the design of powerful imaging experiments that are optimally suited to address their biological questions.