Abstract
Despite much work, subcellular neurons of Caenorhabditis elegans have not been studied at nanometer resolution with millisecond time resolution. Nor has there been an effective way to immobilize C. elegans. Here we show that, without using anesthetic or paralyzing agents, fluorescence imaging with one-nanometer accuracy (FIONA) can be successfully applied to fluorescently labeled molecules within C. elegans nerves. GFP- and DENDRA2-labeled ELKS punctae can be localized with sub-10 nm accuracy in approximately 5 ms. Our results show that the protein ELKS is occasionally transferred by microtubule-based motors. This is the first example of FIONA applied to a living organism.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Animals
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Caenorhabditis elegans Proteins / isolation & purification*
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Caenorhabditis elegans Proteins / metabolism
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Caenorhabditis elegans Proteins / physiology
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Carrier Proteins / isolation & purification*
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Carrier Proteins / metabolism
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Carrier Proteins / physiology
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Cells, Cultured
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Fluorescent Dyes / chemistry
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Green Fluorescent Proteins / chemistry
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Intracellular Signaling Peptides and Proteins
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Kymography
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Molecular Motor Proteins / isolation & purification
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Molecular Motor Proteins / metabolism
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Molecular Motor Proteins / physiology
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Motor Neurons / chemistry
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Motor Neurons / physiology
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Movement / physiology
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Nerve Tissue Proteins / isolation & purification*
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Nerve Tissue Proteins / metabolism
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Nerve Tissue Proteins / physiology
Substances
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Caenorhabditis elegans Proteins
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Carrier Proteins
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Fluorescent Dyes
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Intracellular Signaling Peptides and Proteins
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Molecular Motor Proteins
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Nerve Tissue Proteins
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elks-1 protein, C elegans
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Green Fluorescent Proteins