Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/86003
Title: Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass
Authors: Tie, Hieng Chiong
Chen, Bing
Sun, Xiuping
Cheng, Li
Lu, Lei
Keywords: Golgi
Cellular Biology
Issue Date: 2017
Source: Tie, H. C., Chen, B., Sun, X., Cheng, L., & Lu, L. (2017). Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass. Journal of Visualized Experiments, (126), e55996-.
Series/Report no.: Journal of Visualized Experiments
Abstract: 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. However, the technique and instrument are beyond the capability of most cell biology labs. We describe here our recently developed super-resolution method called Golgi protein localization by imaging centers of mass (GLIM) to systematically and quantitatively localize a Golgi protein. GLIM is based on standard fluorescence labeling protocols and conventional wide-field or confocal microscopes. It involves the calibration of chromatic-shift aberration of the microscopic system, the image acquisition and the post-acquisition analysis. The sub-Golgi localization of a test protein is quantitatively expressed as the localization quotient. There are four main advantages of GLIM; it is rapid, based on conventional methods and tools, the localization result is quantitative, and it affords ~ 30 nm practical resolution along the Golgi axis. Here we describe the detailed protocol of GLIM to localize a test Golgi protein.
URI: https://hdl.handle.net/10356/86003
http://hdl.handle.net/10220/43915
DOI: 10.3791/55996
Schools: School of Biological Sciences 
Rights: © 2017 Journal of Visualized Experiments. This paper was published in Journal of Visualized Experiments and is made available as an electronic reprint (preprint) with permission of Journal of Visualized Experiments. The published version is available at: [http://dx.doi.org/10.3791/55996]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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