Article ; Online: Ultimate use of two-photon fluorescence microscopy to map orientational behavior of fluorophores.
2014 Volume 106, Issue 11, Page(s) 2330–2339
Abstract: ... that polarized two-photon fluorescence based on tunable linear dichroism allows for the retrieval ... The orientational distribution of fluorophores is an important reporter of the structure and ... to the sharpness of the angular constraint, and to the evidence of an isotropic population of fluorophores ...
| Abstract | The orientational distribution of fluorophores is an important reporter of the structure and function of their molecular environment. Although this distribution affects the fluorescence signal under polarized-light excitation, its retrieval is limited to a small number of parameters. Because of this limitation, the need for a geometrical model (cone, Gaussian, etc.) to effect such retrieval is often invoked. In this work, using a symmetry decomposition of the distribution function of the fluorescent molecules, we show that polarized two-photon fluorescence based on tunable linear dichroism allows for the retrieval of this distribution with reasonable fidelity and without invoking either an a priori knowledge of the system to be investigated or a geometrical model. We establish the optimal level of detail to which any distribution can be retrieved using this technique. As applied to artificial lipid vesicles and cell membranes, the ability of this method to identify and quantify specific structural properties that complement the more traditional molecular-order information is demonstrated. In particular, we analyze situations that give access to the sharpness of the angular constraint, and to the evidence of an isotropic population of fluorophores within the focal volume encompassing the membrane. Moreover, this technique has the potential to address complex situations such as the distribution of a tethered membrane protein label in an ordered environment. |
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| MeSH term(s) | Animals ; COS Cells ; Cell Membrane/chemistry ; Chlorocebus aethiops ; Fluorescent Dyes/chemistry ; Liposomes/chemistry ; Microscopy, Fluorescence, Multiphoton/methods ; Models, Chemical ; Pyridinium Compounds/chemistry | ||||||||||
| Chemical Substances | 1-(3-sulfonatopropyl)-4-(beta-(2-(di-n-octylamino)-6-naphthyl)vinyl)pyridinium betaine ; Fluorescent Dyes ; Liposomes ; Pyridinium Compounds | ||||||||||
| Language | English | ||||||||||
| Publishing date | 2014-06-04 | ||||||||||
| Publishing country | United States | ||||||||||
| Document type | Journal Article ; Research Support, Non-U.S. Gov't | ||||||||||
| ZDB-ID | 218078-9 | ||||||||||
| ISSN | 1542-0086 ; 0006-3495 | ||||||||||
| ISSN (online) | 1542-0086 | ||||||||||
| ISSN | 0006-3495 | ||||||||||
| DOI | 10.1016/j.bpj.2014.04.011 | ||||||||||
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| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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