Real time imaging of single fluorophores on moving actin with an epifluorescence microscope

I. Sase, H. Miyata, J. E T Corrie, J. S. Craik, K. Kinosita

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98 Citations (Scopus)

Abstract

Relatively simple modifications of an ordinary epifluorescence microscope have greatly reduced its background luminescence, allowing continuous and real time imaging of single fluorophores in an aqueous medium. Main modifications were changing the excitation light path and setting an aperture stop so that stray light does not scatter inside the microscope. A simple and accurate method using actin filaments is presented to establish the singularity of the observed fluorophores. It was possible, at the video rate of 30 frames/s, to image individual tetramethylrhodamine fluorophores bound to actin filaments sliding over heavy meromyosin. The successful imaging of moving fluorophores demonstrates that conventional microscopes may become a routine tool for studying dynamic interactions among individual biomolecules in physiological environments.

Original languageEnglish
Pages (from-to)323-328
Number of pages6
JournalBiophysical Journal
Volume69
Issue number2
Publication statusPublished - 1995
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics

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  • Cite this

    Sase, I., Miyata, H., Corrie, J. E. T., Craik, J. S., & Kinosita, K. (1995). Real time imaging of single fluorophores on moving actin with an epifluorescence microscope. Biophysical Journal, 69(2), 323-328.