title: High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry.
abstract: The Golgi apparatus is a dynamic organelle, which regulates the vesicular trafficking. While cellular trafficking requires active changes of the Golgi membranes, these are not accompanied by changes in the general Golgi's structure. However, cellular processes such as mitosis, apoptosis and migration require fragmentation of the Golgi complex. Currently, these changes are most commonly studied by basic immunofluorescence and quantified by manual and subjective classification of the Golgi structure in 100-500 stained cells. Several other high-throughput methods exist as well, but those are either complicated or do not provide enough morphological information. Therefore, a simple and informative high content methodology should be beneficial for the study of Golgi architecture. Here we describe the use of high-throughput imaging flow cytometry for quantification of Golgi fragmentation, which provides a simple way to analyze the changes in an automated, quantitative and non-biased manner. Furthermore, it provides a rapid and accurate way to analyze more than 50,000 cells per sample. Our results demonstrate that this method is robust and statistically powerful, thus, providing a much-needed analytical tool for future studies on Golgi dynamics, and can be adapted to other experimental systems.
authors.affiliation: Dept. of Biological Regulation, the Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Inbal
authors.initials: I
authors.lastname: Wortzel
authors.affiliation: Dept. Of Molecular Cell Biology, the Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Koifman
authors.affiliation: Dept. Of Molecular Cell Biology, the Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Varda
authors.initials: V
authors.lastname: Rotter
authors.affiliation: Dept. of Biological Regulation, the Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Rony
authors.initials: R
authors.lastname: Seger
authors.affiliation: Dept. of Life Sciences Core Facilities, the Weizmann Institute of Science, Rehovot, Israel. Ziv.porat@weizmann.ac.il.
authors.forename: Ziv
authors.initials: Z
authors.lastname: Porat
mesh_terms: D000818:Animals; D015415:Biomarkers; D019556:COS Cells; D002522:Chlorocebus aethiops; D005434:Flow Cytometry; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism*; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D051379:Mice; D008938:Mitosis; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology
chemical_list: D015415:Biomarkers