Title: Multiparameter microscopy and spectroscopy for single-molecule analytics
Authors : Prummer, Michael
Sick, Beate
Renn, Alois
Wild, Urs P.
Published in : Analytical Chemistry
Volume(Issue) : 76
Issue : 6
Pages : 1633
Pages to: 1640
Publisher / Ed. Institution : American Chemical Society
Issue Date: 2004
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (publication)
Language : English
Subjects : Optics; Bayes statistics; Single molecule; Multi parameter
Subject (DDC) : 540: Chemistry
Abstract: The ability to monitor several parameters simultaneously from distinct individual fluorescent reporter molecules facilitates the disentanglement of complex and interacting systems and opens new perspectives in areas from basic science to biopharmaceutical technology. By combining annular illumination microscopy, time-correlated singlephoton counting, and multichannel detection, we were able to determine 14 independent parameters from one individual fluorophore. The whole set of parameters was deduced from the few properties of the fluorescence photons, i.e., arrival time, wavelength, and polarization. With this approach, the intensity, the polarization, and the spectral dynamics can be analyzed on a nanosecond time scale and the mean values can be monitored with submillisecond time resolution. Nanosecond spectral dynamics of single molecules has been observed, to the best of our knowledge, for the first time. From our experience, we can determine all parameters for more than 30% of the illuminated fluorophores in biological samples and for more than 80% in doped polymeric films.
Departement: School of Engineering
Organisational Unit: Institute of Data Analysis and Process Design (IDP)
Publication type: Article in scientific journal
DOI : 10.1021/ac034976g
ISSN: 0003-2700
URI: https://digitalcollection.zhaw.ch/handle/11475/14032
Appears in Collections:Publikationen School of Engineering

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