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Microscopy Reference Library

Fluorescent Protein Review Articles

Application of the growing class of fluorescent proteins capable of forming an intrinsic chromophore to the observation of living cells and animals has almost single-handedly launched and fueled a new era in biology and medicine. These powerful research tools have provided investigators with a mechanism of fusing a genetically encoded optical probe to a practically unlimited variety of protein targets in order to examine living systems using fluorescence microscopy and related technology. The references listed below point to review articles that should provide the starting point for a thorough understanding of fluorescent protein technology.

Shaner, N. C., Steinbach, P. A. and Tsien, R. Y.

A guide to choosing fluorescent proteins.  Nature Methods 2: 905-916 (2005).  An excellent reference for researchers attempting to select candidates among the many fluorescent protein color variants for imaging applications using fluorescence and confocal microscopy. The paper includes physical data on fluorescent proteins, partners for multi-color imaging, and filter set recommendations.

Tsien, R. Y.

The green fluorescent protein.  Annual Reviews in Biochemistry 67: 509-545 (1998).  An introduction to the physics, biology, and chemistry of color-shifted Aequorea victoria green fluorescent protein (GFP) variants. Although written over a decade ago, this review article remains among the best sources for gaining an understanding of the fundamental photophysical and genetic properties of fluorescent proteins.

Remington, S. J.

Fluorescent proteins: Maturation, photochemistry and photophysics.  Current Opinion in Structural Biology 16: 714-721 (2006).  A review of the photophysical properties of fluorescent proteins, including a discussion of the various chromophore structures thus far elucidated. The paper also discusses optical highlighters and the reactive oxygen species (ROS) generated by several fluorescent protein variants.

Chudakov, D. M., Lukyanov, S. and Lukyanov, K. A.

Fluorescent proteins as a toolkit for in vivo imaging  Trends in Biotechnology 23: 605-613 (2005).  A complete overview of fluorescent proteins derived from jellyfish and corals, including their use in a wide variety of imaging applications.

Shaner, N. C., Patterson, G. H. and Davidson, M. W.

Advances in fluorescent protein technology.  Journal of Cell Science 120: 4247-4260 (2007).  A review of the best fluorescent proteins in each color class, the engineering methodology underlying the design of new fluorescent protein variants, and optical highlighters.

Dixit, R., Cyr, R. and Gilroy, S.

Using intrinsically fluorescent proteins for plant cell imaging.  The Plant Journal 45: 599-615 (2006).  A thorough investigation of fluorescent protein phototoxicity in plant cells. The authors offer guidelines for imaging that should be applicable to a wide variety of organisms.

Verkhusha, V. V. and Lukyanov, K. A.

The molecular properties and applications of Anthozoa fluorescent proteins and chromoproteins.  Nature Biotechnology 22: 289-296 (2004).  An excellent review of fluorescent proteins derived from Anthozoa corals with comparisons to the jellyfish GFP variants. The authors address spectral diversity, chromophore structure, oligomerization, and applications.

Giepmans, B. N. G., Adams, S. R., Ellisman, M. H. and Tsien, R. Y.

The fluorescent toolbox for assessing protein location and function.  Current Opinion in Structural Biology 16: 714-721 (2006).  Although not limited to fluorescent proteins, this review focuses on the characteristic benefits and limitations of all fluorescent probes used in cell biology. Included are quantum dots, synthetic dyes, fluorescent proteins, immunofluorescence, and small molecule genetically-encoded tags.

Lippincott-Schwartz, J. and Patterson, G. H.

Development and use of fluorescent protein markers in living cells.  Science 300: 87-92 (2003).  An introductory review article outlining the basic properties of fluorescent proteins and optical highlighters with a focus on how these tools are used in cell biology.

Matz, M. V., Lukyanov, K. A. and Lukyanov, S. A.

Family of the green fluorescent protein: journey to the end of the rainbow.  BioEssays 24: 953-959 (2002).  Reviewing the evolutionary aspects of fluorescent proteins, this paper is an excellent source for understanding the diversity of these probes as well as their future possibilities for a variety of investigations ranging from applied biotechnology to evolutionary ecology.