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Live-Cell Imaging of the Estrogen Receptor by Total Internal Reflection Fluorescence Microscopy

Citation

Kisler, K and Dominguez, R, Live-Cell Imaging of the Estrogen Receptor by Total Internal Reflection Fluorescence Microscopy, Estrogen Receptors: Methods and Protocols, Humana Press, KM Eyster (ed), New York, pp. 175-187. ISBN 978-1-4939-3127-9 (2016) [Research Book Chapter]

Copyright Statement

Copyright 2016 Springer Science+Business Media New York

DOI: doi:10.1007/978-1-4939-3127-9_14

Abstract

© Springer Science+Business Media New York 2016. Trafficking studies of plasma membrane-localized intracellular estrogen receptors have mainly relied on biochemical and histological techniques to locate the receptor before and after estradiol stimulation. More often than not these experiments were performed using postmortem, lysed, or fixed tissue samples, whose tissue or cellular structure is typically severely altered or at times completely lost, making the definitive localization of estrogen receptors difficult to ascertain. To overcome this limitation we began using total internal reflection fluorescence microscopy (TIRFM) to study the trafficking of plasma membrane estrogen receptors. This real-time imaging approach, described in this chapter, permits observation of live, intact cells while allowing visualization of the steps (in time and spatial distribution) involved in receptor activation by estradiol and movements on and near the membrane. TIRFM yields high-contrast real-time images of fluorescently labeled E6BSA molecules on and just below the cell surface and is ideal for studying estrogen receptor trafficking in living cells.

Item Details

Item Type:Research Book Chapter
Keywords:Live cell imaging, TIRF, trafficking, estrogen receptor
Research Division:Biological Sciences
Research Group:Zoology
Research Field:Animal Cell and Molecular Biology
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Biological Sciences
Author:Dominguez, R (Dr Reymundo Dominguez)
ID Code:122239
Year Published:2016
Deposited By:Medicine (Discipline)
Deposited On:2017-11-06
Last Modified:2017-11-20
Downloads:0

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