title: High-Content Analysis of the Golgi Complex by Correlative Screening Microscopy.
abstract: The Golgi complex plays a central role in a number of diverse cellular processes, and numerous regulators that control these functions and/or morphology of the Golgi complex are known by now. Many of them were identified by large-scale experiments, such as RNAi-based screening. However, high-throughput experiments frequently provide only initial information that a particular protein might play a role in regulating structure and function of the Golgi complex. Multiple follow-up experiments are necessary to functionally characterize the selected hits. In order to speed up the discovery, we have established a system for correlative screening microscopy that combines rapid data collection and high-resolution imaging in one experiment. We describe here a combination of wide-field microscopy and dual-color direct stochastical optical reconstruction microscopy (dSTORM). We apply the technique to simultaneously capture and differentiate alterations of the cis- and trans-Golgi network when depleting several proteins in a singular and combinatorial manner.
authors.affiliation: BioQuant, University of Heidelberg, 69120, Heidelberg, Germany.
authors.forename: Manuel
authors.initials: M
authors.lastname: Gunkel
authors.affiliation: BioQuant, University of Heidelberg, 69120, Heidelberg, Germany. holger.erfle@bioquant.uni-heidelberg.de.
authors.forename: Holger
authors.initials: H
authors.lastname: Erfle
authors.affiliation: BioQuant, University of Heidelberg, 69120, Heidelberg, Germany.|Department of Biochemistry and Molecular Biology, Faculty of Natural Sciences, Joint Life Sciences Center, University of Vilnius, Vilnius, Lithuania.
authors.forename: Vytaute
authors.initials: V
authors.lastname: Starkuviene
mesh_terms: D006367:HeLa Cells; D006801:Humans; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D021601:trans-Golgi Network / Q000378:metabolism*
keywords: AP3S1; Correlative microscopy; Golgi complex; Golgin-97; dSTORM