The analysis of fluorophore orientation by multiphoton fluorescence microscopy

Jamie M. Leeder; David L. Andrews

doi:10.1117/12.841695

26 February 2010 The analysis of fluorophore orientation by multiphoton fluorescence microscopy

Jamie M. Leeder, David L. Andrews

Proceedings Volume 7569, Multiphoton Microscopy in the Biomedical Sciences X; 756922 (2010) https://doi.org/10.1117/12.841695
Event: SPIE BiOS, 2010, San Francisco, California, United States

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.

Citation Download Citation

Jamie M. Leeder and David L. Andrews "The analysis of fluorophore orientation by multiphoton fluorescence microscopy", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756922 (26 February 2010); https://doi.org/10.1117/12.841695

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available