Identification of single molecules in aqueous solution by time-resolved fluorescence anisotropy

Citation
J. Schaffer et al., Identification of single molecules in aqueous solution by time-resolved fluorescence anisotropy, J PHYS CH A, 103(3), 1999, pp. 331-336
Citations number
26
Categorie Soggetti
Physical Chemistry/Chemical Physics
Journal title
JOURNAL OF PHYSICAL CHEMISTRY A
ISSN journal
10895639 → ACNP
Volume
103
Issue
3
Year of publication
1999
Pages
331 - 336
Database
ISI
SICI code
1089-5639(19990121)103:3<331:IOSMIA>2.0.ZU;2-3
Abstract
Using a confocal epi-illuminated microscope with a polarizing beam splitter and dual-channel detection of single-molecule fluorescence induced by puls ed laser excitation, a new application of the three-dimensional, real-time spectroscopic technique BIFL (burst integrated fluorescence lifetime) is in troduced. BIFL allows simultaneous registration of fluorescence intensity, lifetime, and anisotropy. It is shown to be well-suited to identify the fre ely diffusing fluorescent molecule Rhodamine 123 and the Enhanced Yellow Fl uorescent Protein via their characteristic fluorescence anisotropy using a time-resolved analysis. Furthermore, data analysis is discussed and rotatio nal correlation times of single molecules are determined. Applications for multidimensional single-molecule identification are outlined.