Single molecule polarization microscopy for investigating real time structural dynamics of biological macromolecules

Author

Forkey, Joseph N. ; Quinlan, Margot E. ; Corrie, J.E.T. ; Goldman, Yale E.

Author_Institution

Muscle Inst., Pennsylvania Univ., Philadelphia, PA, USA

Volume

1

fYear

1999

fDate

6/21/1905 12:00:00 AM

Abstract

To quantitatively monitor structural dynamics of individual biomolecules in real time, a single-molecule polarization microscopy technique has been developed. This novel approach combines the single molecule sensitivity of total internal reflection fluorescence microscopy with orientation measurements by time-resolved fluorescence polarization. With this technique, functionally relevant conformational changes of individual biomolecules can be continuously detected at 10 ms time resolution or better. Initial measurements, using site specific fluorescently labeled skeletal muscle myosin showed changes in the orientation of the light chain domain, which has been postulated to act as a lever arm during chemo-mechanical energy transduction

Keywords

biological techniques; fluorescence; light polarisation; macromolecules; molecular biophysics; muscle; optical microscopy; proteins; biological macromolecules; chemo-mechanical energy transduction; functionally relevant conformational changes; individual biomolecules; lever arm; light chain domain; orientation; real time structural dynamics; single-molecule polarization microscopy technique; site specific fluorescently labeled skeletal muscle myosin; Energy measurement; Energy resolution; Fluorescence; Microscopy; Molecular biophysics; Monitoring; Muscles; Optical polarization; Optical reflection; Position measurement;

fLanguage

English

Publisher

ieee

Conference_Titel

[Engineering in Medicine and Biology, 1999. 21st Annual Conference and the 1999 Annual Fall Meetring of the Biomedical Engineering Society] BMES/EMBS Conference, 1999. Proceedings of the First Joint

ISSN

1094-687X

Print_ISBN

0-7803-5674-8

Type

conf

DOI

10.1109/IEMBS.1999.802112

Filename

802112