title: Comparative assessment of fluorescent proteins for in vivo imaging in an animal model system.
abstract: Fluorescent protein tags are fundamental tools used to visualize gene products and analyze their dynamics in vivo. Recent advances in genome editing have expedited the precise insertion of fluorescent protein tags into the genomes of diverse organisms. These advances expand the potential of in vivo imaging experiments and facilitate experimentation with new, bright, photostable fluorescent proteins. Most quantitative comparisons of the brightness and photostability of different fluorescent proteins have been made in vitro, removed from biological variables that govern their performance in cells or organisms. To address the gap, we quantitatively assessed fluorescent protein properties in vivo in an animal model system. We generated transgenic Caenorhabditis elegans strains expressing green, yellow, or red fluorescent proteins in embryos and imaged embryos expressing different fluorescent proteins under the same conditions for direct comparison. We found that mNeonGreen was not as bright in vivo as predicted based on in vitro data but is a better tag than GFP for specific kinds of experiments, and we report on optimal red fluorescent proteins. These results identify ideal fluorescent proteins for imaging in vivo in C. elegans embryos and suggest good candidate fluorescent proteins to test in other animal model systems for in vivo imaging experiments.
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
authors.forename: Jennifer K
authors.initials: JK
authors.lastname: Heppert
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.|Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Dickinson
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.|Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
authors.forename: Ariel M
authors.initials: AM
authors.lastname: Pani
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Higgins
authors.affiliation: Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, United Kingdom.
authors.forename: Annette
authors.initials: A
authors.lastname: Steward
authors.affiliation: Gurdon Institute, University of Cambridge, Cambridge CB2 1QN, United Kingdom.
authors.forename: Julie
authors.initials: J
authors.lastname: Ahringer
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Kuhn
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 bobg@unc.edu.|Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
authors.forename: Bob
authors.initials: B
authors.lastname: Goldstein
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism; D004195:Disease Models, Animal; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D061848:Optical Imaging / Q000379:methods*; D000097573:Red Fluorescent Protein
chemical_list: D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein