Three-dimensional orientation of single molecules observed by far- and near-field fluorescence microscopy

We present a simple and straightforward method for determining absolute spatial orientations of transition dipole moments of single fluorescent molecules. Far-field polarization microscopy provides angles of the dipole moments projected in the plane of the sample. Optical field near total internal r...

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Bibliographic Details
Published in:The Journal of chemical physics 2003-03, Vol.118 (12), p.5279-5282
Main Authors: Vacha, Martin, Kotani, Masahiro
Format: Article
Language:English
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Summary:We present a simple and straightforward method for determining absolute spatial orientations of transition dipole moments of single fluorescent molecules. Far-field polarization microscopy provides angles of the dipole moments projected in the plane of the sample. Optical field near total internal reflection surfaces has a strong component perpendicular to the sample and, for a given in-plane angle, provides unambiguous orientation of the molecular dipole moment. Experimentally, both excitation modes are alternated to monitor real-time conformational dynamics of tetramethylrhodamine molecules covalently attached to a quartz substrate.
ISSN:0021-9606
1089-7690
DOI:10.1063/1.1562194