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

Authors:

Agnes Koerfer, Francesco Reina, Christian Eggeling

Keywords:

STED-FCS, surface cleaning, imaging

Abstract:

Molecular 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.

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