TIE Hieng Chiong

Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass

Published date : 08 Oct 2017

The Golgi complex consists of serially stacked membrane cisternae which can be further categorized into sub-Golgi regions, including the cis-Golgi, medial-Golgi, trans-Golgi and trans-Golgi network. Cellular functions of the Golgi are determined by the characteristic distribution of its resident proteins. The spatial resolution of conventional light microscopy is too low to resolve sub-Golgi structure or cisternae. Thus, the immuno-gold electron microscopy is a method of choice to localize a protein at the sub-Golgi level.

type
Journal Paper
journal
JoVE Journal of Visualized Experiments, 2017, doi:10.3791/55996

A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes

Published date : 13 Jan 2016

Cellular functions of the Golgi are determined by the unique distribution of its resident proteins. Currently, electron microscopy is required for the localization of a Golgi protein at the sub-Golgi level. We developed a quantitative sub-Golgi localization method based on centers of fluorescence masses of nocodazole-induced Golgi ministacks under conventional optical microscopy. Our method is rapid, convenient, and quantitative, and it yields a practical localization resolution of ∼30 nm. The method was validated by the previous electron microscopy data.

type
Journal Paper
journal
Molecular Biology Of The Cell, Vol. 27, No. 5, Mar 1 2016, pg 848-861, doi: 10.1091/mbc
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