The analysis of fluorophore orientation by multiphoton fluorescence microscopy

Leeder, J.M. and Andrews, D.L. (2010) The analysis of fluorophore orientation by multiphoton fluorescence microscopy. In: Progress in Biomedical Optics and Imaging - Proceedings of SPIE. UNSPECIFIED. ISBN 9780819479655

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Abstract

The accessibility of tunable, ultrafast laser sources has spurred the development and wide application of specialized microscopy techniques based on chromophore fluorescence following two- and three-photon absorption. The attendant advantages of such methods, which have led to a host of important applications including three-dimensional biological imaging, include some features that have as yet received relatively little attention. In the investigation of cellular or subcellular processes, it is possible to discern not only on the location, concentration, and lifetime of molecular species, but also the orientations of key fluorophores. Detailed information can be secured on the degree of orientational order in specific cellular domains, or the lifetimes associated with the rotational motions of individual fluorophores; both are accessible from polarization-resolved measurements. This paper reports the equations that are required for any such investigation, determined by robust quantum electrodynamical derivation. The general analysis, addressing a system of chromophores oriented in three dimensions, determines the fluorescence signal produced by the nonlinear polarizations that are induced by multiphoton absorption, allowing for any rotational relaxation. The results indicate that multiphoton imaging can be further developed as a diagnostic tool, either to selectively discriminate micro-domains in vivo, or to monitor dynamical changes in intracellular fluorophore orientation.

Item Type: Book Section
Uncontrolled Keywords: multiphoton fluorescence microscopy,absorption,luminescence,microscopy,molecules,diagnostics,laser sources,photons,polarization
Faculty \ School: Faculty of Science > School of Chemistry
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Depositing User: Pure Connector
Date Deposited: 12 May 2014 15:18
Last Modified: 22 Apr 2020 10:42
URI: https://ueaeprints.uea.ac.uk/id/eprint/48229
DOI: 10.1117/12.841695

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