A quantitative fluorescence-imaging technique for studying acetylcholine receptor turnover at neuromuscular junctions in living animals

Stephen G. Turney, Susan M. Culican, Jeff W. Lichtman

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

We have developed a technique to measure changes in the amount of fluorescently labeled acetylcholine receptors in living muscles over long time periods. The measurements of fluorescence are made relative to a novel, photolytically stable fluorescence standard (Spectralon) which allows changes in fluorescence to be followed over days, even months. The method compensates for spatial and temporal variations in image brightness due to the light source, microscope, and camera. We use this approach to study the turnover of fluorescently labeled acetylcholine receptors at a single neuromuscular junction in a living mouse by re-imaging the same junction in situ over a period of 3 weeks. In addition we show that the SIT video camera, which is generally considered inadequate for quantitative imaging (in comparison to CCD cameras), is actually a very good quantitative device, especially in situations requiring both fast acquisition and high resolution.

Original languageEnglish (US)
Pages (from-to)199-208
Number of pages10
JournalJournal of Neuroscience Methods
Volume64
Issue number2
DOIs
StatePublished - Feb 1996
Externally publishedYes

Keywords

  • Acetylcholine receptor
  • Fluorescence standard
  • Microfluorometry
  • Neuromuscular junction
  • Quantitative fluorescence microscopy
  • SIT video camera
  • Spectralon
  • α-Bungarotoxin

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