Subdiffraction resolution microscopy methods for analyzing centrosomes organization

Vito Mennella; Rachel Hanna; Moshe Kim

doi:10.1016/bs.mcb.2015.03.009

Subdiffraction resolution microscopy methods for analyzing centrosomes organization

Methods Cell Biol. 2015:129:129-152. doi: 10.1016/bs.mcb.2015.03.009. Epub 2015 May 27.

Authors

Vito Mennella¹, Rachel Hanna², Moshe Kim²

Affiliations

¹ Department of Biochemistry, University of Toronto, Toronto, ON, Canada; Cell Biology Program, The Hospital for Sick Children, Toronto, ON, Canada; Peter Gilgan Centre for Research and Learning, Toronto, ON, Canada.
² Department of Biochemistry, University of Toronto, Toronto, ON, Canada; Cell Biology Program, The Hospital for Sick Children, Toronto, ON, Canada.

PMID: 26175437
DOI: 10.1016/bs.mcb.2015.03.009

Abstract

In this chapter, we describe the current methods of examining the structure of centrosomes by fluorescence subdiffraction microscopy. By using recently developed microscopy techniques, centrosomal proteins can now be examined in cells with a resolution of only a few nanometers, a level of molecular detail beyond the reach of traditional cell biology methods as confocal and widefield microscopy. We emphasize imaging by three-dimensional structured illumination microscopy, stochastic optical reconstruction microscopy, and quantitative approaches to image data analysis.

Keywords: 3DSIM; Averaging; Centrosome structure; Fluorescence light microscopy; Macromolecular assembly; PCM; STORM; Subdiffraction imaging; Superresolution imaging.

MeSH terms

Animals
Cell Line
Centrosome / physiology
Centrosome / ultrastructure*
Drosophila melanogaster
Fluorescent Antibody Technique, Indirect
Microscopy, Fluorescence / methods