High-Content Analysis of the Golgi Complex by Correlative Screening Microscopy.

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.

Citation

Manuel Gunkel, Holger Erfle, Vytaute Starkuviene. High-Content Analysis of the Golgi Complex by Correlative Screening Microscopy. Methods in molecular biology (Clifton, N.J.). 2016;1496:111-21

PMID: 27632005

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