Advanced

Quantitative measurement of fluorescence brightness of single molecules

Tian, Yuxi LU ; Halle, Johannes; Wojdyr, Michal; Sahoo, Dibakar LU and Scheblykin, Ivan LU (2014) In Methods and Applications in Fluorescence 2(3).
Abstract
Single-molecule fluorescence spectroscopy and imaging probe many characteristics of the fluorescence from individual molecules like relative intensity, polarization, lifetime and spectrum. However, such an important and fundamental parameter as absolute fluorescence intensity (or in other words fluorescence brightness), which is proportional to the absorption cross section and fluorescence quantum yield, has not yet been sufficiently exploited in the field. One reason for that is the difficulty of absolute fluorescence brightness measurements. In the present work a detailed description of fluorescence brightness measurements of single molecules is given. We discuss several important factors like the power density and polarization of... (More)
Single-molecule fluorescence spectroscopy and imaging probe many characteristics of the fluorescence from individual molecules like relative intensity, polarization, lifetime and spectrum. However, such an important and fundamental parameter as absolute fluorescence intensity (or in other words fluorescence brightness), which is proportional to the absorption cross section and fluorescence quantum yield, has not yet been sufficiently exploited in the field. One reason for that is the difficulty of absolute fluorescence brightness measurements. In the present work a detailed description of fluorescence brightness measurements of single molecules is given. We discuss several important factors like the power density and polarization of excitation light, the substrates and the local environment. It is shown that the fluorescence brightness of a single molecule indeed can be measured with sufficient accuracy and used as a powerful parameter for characterization of materials at single molecule/particle level. The brightness of a single object can give similar information as the fluorescence quantum yield that is crucial for understanding the photophysical properties for individual multi-chromophoric systems in inhomogeneous environments. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
single molecule detection, brightness, fluorescence quantum yield, excitation power, polarization
in
Methods and Applications in Fluorescence
volume
2
issue
3
publisher
IOP Publishing
external identifiers
  • wos:000353362000004
  • scopus:84974622089
ISSN
2050-6120
DOI
10.1088/2050-6120/2/3/035003
language
English
LU publication?
yes
id
97f60cc6-ca07-400d-886c-3ce580ddbdb3 (old id 5401197)
date added to LUP
2015-05-19 11:19:47
date last changed
2017-09-24 04:07:55
@article{97f60cc6-ca07-400d-886c-3ce580ddbdb3,
  abstract     = {Single-molecule fluorescence spectroscopy and imaging probe many characteristics of the fluorescence from individual molecules like relative intensity, polarization, lifetime and spectrum. However, such an important and fundamental parameter as absolute fluorescence intensity (or in other words fluorescence brightness), which is proportional to the absorption cross section and fluorescence quantum yield, has not yet been sufficiently exploited in the field. One reason for that is the difficulty of absolute fluorescence brightness measurements. In the present work a detailed description of fluorescence brightness measurements of single molecules is given. We discuss several important factors like the power density and polarization of excitation light, the substrates and the local environment. It is shown that the fluorescence brightness of a single molecule indeed can be measured with sufficient accuracy and used as a powerful parameter for characterization of materials at single molecule/particle level. The brightness of a single object can give similar information as the fluorescence quantum yield that is crucial for understanding the photophysical properties for individual multi-chromophoric systems in inhomogeneous environments.},
  articleno    = {035003},
  author       = {Tian, Yuxi and Halle, Johannes and Wojdyr, Michal and Sahoo, Dibakar and Scheblykin, Ivan},
  issn         = {2050-6120},
  keyword      = {single molecule detection,brightness,fluorescence quantum yield,excitation power,polarization},
  language     = {eng},
  number       = {3},
  publisher    = {IOP Publishing},
  series       = {Methods and Applications in Fluorescence},
  title        = {Quantitative measurement of fluorescence brightness of single molecules},
  url          = {http://dx.doi.org/10.1088/2050-6120/2/3/035003},
  volume       = {2},
  year         = {2014},
}