Work

Estimating the size and number of autophagic bodies by electron microscopy

Year: 2013

Type: article

Abstract: Much recent and ongoing research is focused on understanding the mechanisms and regulation of autophagy, a cellular self-degradation pathway with many links to human health. Although many assays exist to measure the total magnitude of autophagy, electron microscopy remains the tool of choice for the determination of the size and the number of autophagosomes formed in a given mutant or under given induction conditions. Here we present a detailed protocol for measuring autophagic bodies in the yeast Saccharomyces cerevisiae by electron microscopy. Furthermore, we present an improved mathematical method for estimating body size and a new method for estimating body number. Finally, we include a discussion of the merits and limitations of these methods and an example of their application to autophagic bodies formed in the ume6∆ strain.

Source: Autophagy

DOI: 10.4161/auto.26856

Authors: Steven K Backues, Dachuan Chen, Jishou Ruan, Zhiping Xie, Daniel J Klionsky

Institutions: University of Michigan, Life Sciences Institute, Nankai University, State Key Laboratory of Medicinal Chemical Biology

Language: English

Topic: Autophagy in Disease and Therapy

Subfield: Epidemiology

Field: Medicine

Domain: Health Sciences

Open Access status: bronze