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Quantitative measurement of fluorescence brightness of single molecules

Tian, Yuxi LU ; Halle, Johannes ; Wojdyr, Michal ; Sahoo, Dibakar LU and Scheblykin, Ivan LU orcid (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)
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author
; ; ; and
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
article number
035003
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
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Chemical Physics (S) (011001060)
id
97f60cc6-ca07-400d-886c-3ce580ddbdb3 (old id 5401197)
date added to LUP
2016-04-01 14:23:32
date last changed
2021-10-10 04:03:02
@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.},
  author       = {Tian, Yuxi and Halle, Johannes and Wojdyr, Michal and Sahoo, Dibakar and Scheblykin, Ivan},
  issn         = {2050-6120},
  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},
  doi          = {10.1088/2050-6120/2/3/035003},
  volume       = {2},
  year         = {2014},
}