Simultaneous multicolor fluorescence cross-correlation spectroscopy to detect higher order molecular interactions using single wavelength laser excitation.

Biophysical Journal
Ling Chin HwangThorsten Wohland

Abstract

Fluorescence cross-correlation spectroscopy is a powerful method for the study of molecular interactions and dynamics in solution and even in living cells. Usually, in the optical setup, either two laser beams have to be superimposed in their respective confocal volumes or two-photon excitation is used for a dual-color detection system. It has been shown recently that fluorescence cross correlation can be achieved with spectrally similar fluorophores using single wavelength excitation fluorescence cross-correlation spectroscopy (SW-FCCS). In this study, we show that SW-FCCS allows the simultaneous excitation of up to three fluorophores in which the cross correlation of their fluctuation signals is detected separately in three detection channels. The experimental and theoretical model to describe triple pairwise cross correlations incorporating cross talk and possible changes in emission characteristics such as quenching upon binding are outlined. The effectiveness of SW-FCCS to detect binding of three interacting partners is experimentally verified with a standard ligand-receptor model, biotin-streptavidin, where differently labeled biotin ligands and their binding to a third-color labeled streptavidin are studied. The cross-co...Continue Reading

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Citations

Sep 18, 2007·Cell Biochemistry and Biophysics·Ling Chin Hwang, Thorsten Wohland
Apr 11, 2012·Journal of the American Chemical Society·Daria M ShcherbakovaVladislav V Verkhusha
Jan 5, 2011·The Journal of Physical Chemistry. B·Joseph P SkinnerSergey Y Tetin
Aug 8, 2012·Proceedings of the National Academy of Sciences of the United States of America·William K RidgewayJames R Williamson
May 5, 2007·Annual Review of Biophysics and Biomolecular Structure·Elke Haustein, Petra Schwille
Jun 14, 2013·The Journal of Chemical Physics·Ligang FengHans-Jürgen Butt
Apr 5, 2008·Trends in Endocrinology and Metabolism : TEM·Ping LiuThorsten Wohland
Aug 27, 2013·Biophysical Journal·Jelle HendrixDon C Lamb
Jun 17, 2009·Biophysical Journal·Daniel J NeedlemanTimothy J Mitchison
May 18, 2016·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Hernán E GreccoEli Zamir
Jun 8, 2007·The Review of Scientific Instruments·Xiaotao PanThorsten Wohland
Apr 21, 2018·Integrative Biology : Quantitative Biosciences From Nano to Macro·Bianca K StöckerSven Rahmann
Jun 10, 2010·Analytical Chemistry·Stefan Wennmalm, Jerker Widengren
Mar 7, 2012·Journal of Cell Science·Jiji ChenJoseph M K Irudayaraj
Jan 25, 2018·Chemical Reviews·Sujay RayNils G Walter

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