Ex-dSTORM and automated quantitative image analysis of expanded filamentous structures

Fabian U Zwettler; Sebastian Reinhard; Markus Sauer

doi:10.1016/bs.mcb.2020.05.004

Ex-dSTORM and automated quantitative image analysis of expanded filamentous structures

Methods Cell Biol. 2021:161:317-340. doi: 10.1016/bs.mcb.2020.05.004. Epub 2020 Jun 10.

Authors

Fabian U Zwettler¹, Sebastian Reinhard¹, Markus Sauer²

Affiliations

¹ Department of Biotechnology and Biophysics, Biocenter, University of Würzburg, Würzburg, Germany.
² Department of Biotechnology and Biophysics, Biocenter, University of Würzburg, Würzburg, Germany. Electronic address: m.sauer@uni-wuerzburg.de.

PMID: 33478695
DOI: 10.1016/bs.mcb.2020.05.004

Abstract

This chapter provides a step-by-step protocol how to prepare expansion microcoscopy (ExM) treated biological samples for imaging with single-molecule localization microscopy (SMLM). For this purpose, the protocol describes the stabilization of expanded hydrogels that enables addition of photoswitching buffer without shrinkage of the sample. In addition, a guide for automated image analysis and expansion factor determination of expanded fiber-like structures is provided at the end of the chapter.

Keywords: Expansion microscopy; LineProfiler; Super-resolution microscopy; dSTORM.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Image Processing, Computer-Assisted*
Single Molecule Imaging*