Abstract

The multifrequency phase and modulation (MPM) technique is used in concert with total internal reflection fluorescence (TIRF) to probe <i>in situ</i> the distribution of free and analyte-bound sites of a fluorescently labeled, silica-immobilized <i>F</i>_(ab') antibody fragment. Using this new technique, one is able to recover the affinity constant (<i>K</i>_<i>f</i>) for the immobilized antibody, determine the effects of storage time on <i>K</i>_<i>f</i> and the excited-state fluorescence decay parameters, and follow time-dependent changes in antibody conformation at the silica substrata. This new technique adds a powerful tool to the study of biosensor interfaces.

Fluorescence correlation spectroscopy with a total internal reflection fluorescence STED microscope (TIRF-STED-FCS)

Marcel Leutenegger, Christian Ringemann, Theo Lasser, Stefan W. Hell, and Christian Eggeling
Opt. Express 20(5) 5243-5263 (2012)

Around-the-objective total internal reflection fluorescence microscopy

Thomas P. Burghardt, Andrew D. Hipp, and Katalin Ajtai
Appl. Opt. 48(32) 6120-6131 (2009)

Phase shifts that accompany total internal reflection at a dielectric–dielectric interface

R. M. A. Azzam
J. Opt. Soc. Am. A 21(8) 1559-1563 (2004)

Absorption effects on total-internal-reflection fluorescence spectroscopy

Minoru Toriumi, Masatoshi Yanagimachi, and Hiroshi Masuhara
Appl. Opt. 31(30) 6376-6382 (1992)

Total-internal-reflection fluorescence microscopy with W-shaped axicon mirrors

Ming Lei and Andreas Zumbusch
Opt. Lett. 35(23) 4057-4059 (2010)