Related Experiment Video
Updated: Jun 4, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
Published on: October 28, 2018
Mechanisms and advancement of antifading agents for fluorescence microscopy and single-molecule spectroscopy
Thorben Cordes1, Andreas Maiser, Christian Steinhauer
1Applied Physics-Biophysics & Center for NanoScience (CeNS), Ludwig Maximilian University of Munich, Amalienstr. 54, 80799 Munich, Germany. thorben.cordes@physik.lmu.de
Abstract:
Modern fluorescence microscopy applications go along with increasing demands for the employed fluorescent dyes. In this work, we compared antifading formulae utilizing a recently developed reducing and oxidizing system (ROXS) with commercial antifading agents. To systematically test fluorophore performance in fluorescence imaging of biological samples, we carried out photobleaching experiments using fixed cells labeled with various commonly used organic dyes, such as Alexa 488, Alexa 594, Alexa 647, Cy3B, ATTO 550, and ATTO 647N. Quantitative evaluation of (i) photostability, (ii) brightness, and (iii) storage stability of fluorophores in samples mounted in different antifades (AFs) reveal optimal combinations of dyes and AFs. Based on these results we provide guidance on which AF should preferably be used with a specific dye. Finally, we studied the antifading mechanisms of the commercial AFs using single-molecule spectroscopy and reveal that these empirically selected AFs exhibit similar properties to ROXS AFs.
Related Concept Videos
Super-resolution Fluorescence Microscopy
Protein Dynamics in Living Cells
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Two-Dimensional Microscopy in Microbiology
Fluorescence and Phosphorescence: Instrumentation

