Influence of different surface cleaning methods on STED‐FCS and scanning STED‐FCS calibration measurements

Author:

Koerfer Agnes12,Reina Francesco12,Eggeling Christian123ORCID

Affiliation:

1. Institute of Applied Optics and Biophysics Friedrich Schiller University Jena Jena Germany

2. Leibniz Institute of Photonic Technology eV, member of the Leibniz Centre for Photonics in Infection Research (LPI) Jena Germany

3. Jena Center for Soft Matter (JCSM) Jena Germany

Abstract

AbstractMolecular mobility is an important measure in biological functionality, as molecules have to diffuse to meet and interact and perform actions. Measurement of mobility requires specific tools such as fluorescence correlation spectroscopy (FCS). Especially, combination with superresolution stimulated emission depletion microscopy (STED‐FCS), whether in a point‐ or beam‐scanning mode, has proven valuable for determination of anomalous diffusion. STED‐FCS however relies on an accurate calibration of the effective observation spot formed for different laser powers of the additional STED laser. This poster article highlights the need for calibration measurements and outlines that rather simple procedures involving acetone cover‐glass surface cleaning only, instead of piranha cover‐glass surface cleaning, and point instead of more complex scanning STED‐FCS are sufficient for calibration.

Funder

Deutsche Forschungsgemeinschaft

Bundesministerium für Bildung und Frauen

Publisher

Wiley

Subject

Histology,Pathology and Forensic Medicine

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