title: Quantitative assessment of fluorescent proteins.
abstract: The advent of fluorescent proteins (FPs) for genetic labeling of molecules and cells has revolutionized fluorescence microscopy. Genetic manipulations have created a vast array of bright and stable FPs spanning blue to red spectral regions. Common to autofluorescent FPs is their tight β-barrel structure, which provides the rigidity and chemical environment needed for effectual fluorescence. Despite the common structure, each FP has unique properties. Thus, there is no single 'best' FP for every circumstance, and each FP has advantages and disadvantages. To guide decisions about which FP is right for a given application, we have quantitatively characterized the brightness, photostability, pH stability and monomeric properties of more than 40 FPs to enable straightforward and direct comparison between them. We focus on popular and/or top-performing FPs in each spectral region.
authors.affiliation: National High Field Magnet Lab, Florida State University, Tallahassee, Florida, USA.|Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, USA.
authors.forename: Paula J
authors.initials: PJ
authors.lastname: Cranfill
authors.affiliation: National High Field Magnet Lab, Florida State University, Tallahassee, Florida, USA.
authors.forename: Brittney R
authors.initials: BR
authors.lastname: Sell
authors.affiliation: National High Field Magnet Lab, Florida State University, Tallahassee, Florida, USA.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Baird
authors.affiliation: National High Field Magnet Lab, Florida State University, Tallahassee, Florida, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Allen
authors.affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, USA.|Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri, USA.
authors.forename: Zeno
authors.initials: Z
authors.lastname: Lavagnino
authors.affiliation: Erasmus Optical Imaging Centre, Josephine Nefkens Institute, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: H Martijn
authors.initials: HM
authors.lastname: de Gruiter
authors.affiliation: Erasmus Optical Imaging Centre, Josephine Nefkens Institute, Erasmus MC, Rotterdam, the Netherlands.
authors.forename: Gert-Jan
authors.initials: GJ
authors.lastname: Kremers
authors.affiliation: National High Field Magnet Lab, Florida State University, Tallahassee, Florida, USA.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Davidson
authors.affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, USA.|Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri, USA.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ustione
authors.affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee, USA.|Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri, USA.
authors.forename: David W
authors.initials: DW
authors.lastname: Piston
mesh_terms: D005453:Fluorescence; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis*; D008856:Microscopy, Fluorescence / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000032:analysis*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins