Book Chapter

Application of flow cytometry to analyze intracellular location and trafficking of cargo in cell populations

WH Toh, FJ Houghton, CP Chia, YM Ramdzan, DM Hatters, PA Gleeson

Membrane Trafficking: Second Edition | Methods in Molecular Biology | Humana Press | Published : 2015

Abstract

© Springer Science+Business Media New York 2015. All rights are reserved.Pulse shape analysis (PulSA) is a flow cytometry-based method that involves the measurement of the pulse width and height of a fluorescently labeled molecule simultaneously, enabling a multidimensional analysis of protein localization in a cell at high speed and throughput. We have used the method to detect morphological changes in organelles such as Golgi fragmentation, track protein trafficking from the cell surface, and also discriminate cells with different target protein localizations such as the Golgi, lyso-endosomal network, and the plasma membrane. Here, we describe the basic experimental setup and analytical me..

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