PMID: 9450329Feb 5, 1998Paper

Dual-colour microscopy of single fluorophores bound to myosin interacting with fluorescently labelled actin using anti-Stokes fluorescence

Journal of Microscopy
K SaitoT Yanagida

Abstract

We have refined prismless total internal reflection fluorescence microscopy with extremely low background to visualize single fluorophores attached to protein molecules interacting with a filamentous biopolymer labelled with different colour fluorophores. By using Stokes and anti-Stokes fluorescence, two different colour fluorescences from two different colour fluorophores excited with a single wavelength laser can be observed simultaneously. This microscopy was applied to visualize motor proteins, actin and myosin molecules. Single myosin molecules labelled with a tetramethylrhodamine-5-iodoacetamide interacting with a BODIPY FL-labelled actin filament, a filamentous polymer of actin molecules, were observed clearly and simultaneously in aqueous solution. Individual hydrolysis reactions of Cy3-labelled ATP by single myosin molecules and sliding of a BODIPY FL-labelled actin filament along the myosin molecules could also be observed simultaneously. Thus, this technique is useful for observing single molecular processes of proteins interacting with a biological macromolecule such as an actin filament and a DNA.

Citations

Feb 1, 2003·Experimental Gerontology·G J Schütz, P Hinterdorfer
Feb 19, 2000·Current Opinion in Cell Biology·T YanagidaS Esaki
May 10, 2002·Journal of Biotechnology·P HinterdorferH Schindler
Nov 30, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jan HesseGerhard J Schütz
Jan 3, 2009·Methods in Cell Biology·Daniel Axelrod
May 29, 2000·Molecular Membrane Biology·G J SchützH Schindler
Nov 22, 2018·Applied Optics·Abdollah HassanzadehSalah Raza Saeed
Dec 26, 2001·Chemical Reviews·W P AmbroseR A Keller

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