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Citation
Viswanathan, S., Williams, M.E., Bloss, E.B., Stasevich, T.J., Speer, C.M., Nern, A., Pfeiffer, B.D., Hooks, B.M., Li, W.P., English, B.P., Tian, T., Henry, G.L., Macklin, J.J., Patel, R., Gerfen, C.R., Zhuang, X., Wang, Y., Rubin, G.M., Looger, L.L. (2015). High-performance probes for light and electron microscopy.  Nat. Methods 12(6): 568--576.
FlyBase ID
FBrf0229229
Publication Type
Research paper
Abstract

We describe an engineered family of highly antigenic molecules based on GFP-like fluorescent proteins. These molecules contain numerous copies of peptide epitopes and simultaneously bind IgG antibodies at each location. These 'spaghetti monster' fluorescent proteins (smFPs) distributed well in neurons, notably into small dendrites, spines and axons. smFP immunolabeling localized weakly expressed proteins not well resolved with traditional epitope tags. By varying epitope and scaffold, we generated a diverse family of mutually orthogonal antigens. In cultured neurons and mouse and fly brains, smFP probes allowed robust, orthogonal multicolor visualization of proteins, cell populations and neuropil. smFP variants complement existing tracers and greatly increase the number of simultaneous imaging channels, and they performed well in advanced preparations such as array tomography, super-resolution fluorescence imaging and electron microscopy. In living cells, the probes improved single-molecule image tracking and increased yield for RNA-seq. These probes facilitate new experiments in connectomics, transcriptomics and protein localization.

PubMed ID
PubMed Central ID
PMC4573404 (PMC) (EuropePMC)
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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    Nat. Methods
    Title
    Nature Methods
    Publication Year
    2004-
    ISBN/ISSN
    1548-7091 1548-7105
    Data From Reference