FIONA on Caenorhabditis elegans.
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.
Kural
Comert
C
Center for Biophysics and Computational Biology, University of Illinois, Urbana, Illinois 61801, USA.
Nonet
Michael L
ML
Selvin
Paul R
PR
D000818:Animals; D029742:Caenorhabditis elegans Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology; D002478:Cells, Cultured; D005456:Fluorescent Dyes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D047908:Intracellular Signaling Peptides and Proteins; D007734:Kymography; D020409:Molecular Motor Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D009046:Motor Neurons / Q000737:chemistry / Q000502:physiology; D009068:Movement / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology
D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D005456:Fluorescent Dyes; D047908:Intracellular Signaling Peptides and Proteins; D020409:Molecular Motor Proteins; D009419:Nerve Tissue Proteins; C508059:elks-1 protein, C elegans; D049452:Green Fluorescent Proteins