Epifluorescence, confocal and total internal reflection microscopy for single-molecule experiments: a quantitative comparison
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Updated: Aug 7, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
Published on: October 28, 2018
1Experimental Physics IV and BIMF, University of Bayreuth, 95440 Bayreuth, Germany.
Abstract:
Epifluorescence, confocal and total internal reflection microscopy are the most widely used techniques for optical single-molecule experiments. Employing these methods, we recorded the emission intensity of the same single molecule as a function of the excitation rate under otherwise identical experimental conditions. Evaluation of these data provides a quantitative comparison of the signal-to-background ratios that can be achieved for the three microscopic techniques.
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