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...

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Autores principales:	Inbal Wortzel, Gabriela Koifman, Varda Rotter, Rony Seger, Ziv Porat
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4796301aa11b4508800e2f0e548c2abb
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spelling	oai:doaj.org-article:4796301aa11b4508800e2f0e548c2abb2021-12-02T12:32:16ZHigh Throughput Analysis of Golgi Structure by Imaging Flow Cytometry10.1038/s41598-017-00909-y2045-2322https://doaj.org/article/4796301aa11b4508800e2f0e548c2abb2017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00909-yhttps://doaj.org/toc/2045-2322Abstract 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.Inbal WortzelGabriela KoifmanVarda RotterRony SegerZiv PoratNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Inbal Wortzel Gabriela Koifman Varda Rotter Rony Seger Ziv Porat High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry
description	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.
format	article
author	Inbal Wortzel Gabriela Koifman Varda Rotter Rony Seger Ziv Porat
author_facet	Inbal Wortzel Gabriela Koifman Varda Rotter Rony Seger Ziv Porat
author_sort	Inbal Wortzel
title	High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry
title_short	High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry
title_full	High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry
title_fullStr	High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry
title_full_unstemmed	High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry
title_sort	high throughput analysis of golgi structure by imaging flow cytometry
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/4796301aa11b4508800e2f0e548c2abb
work_keys_str_mv	AT inbalwortzel highthroughputanalysisofgolgistructurebyimagingflowcytometry AT gabrielakoifman highthroughputanalysisofgolgistructurebyimagingflowcytometry AT vardarotter highthroughputanalysisofgolgistructurebyimagingflowcytometry AT ronyseger highthroughputanalysisofgolgistructurebyimagingflowcytometry AT zivporat highthroughputanalysisofgolgistructurebyimagingflowcytometry
_version_	1718394091162566656

High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry

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