Journal of Fluorescence

, Volume 20, Issue 1, pp 401–405 | Cite as

Direct Visualization of Vesicle Maturation and Plasma Membrane Protein Trafficking

Short Communication


Internalization and intracellular trafficking of membrane proteins are now recognized as essential mechanisms that contribute to a number of cellular processes. Current methods lack the ability to specifically label the plasma membrane of a live cell, follow internalization of labeled membrane molecules, and conclusively differentiate newly formed membrane-derived vesicles from pre-existing endocytic or secretory structures in the cytoplasm. Here, we detail a visualization method for surface biotinylation of plasma membrane-derived vesicles that allows us to follow their progress from membrane to cytosol at specific time points. Using the transmembrane receptor RET as a model, we demonstrate how this method can be applied to identify plasma membrane-derived vesicle maturation, determine RET’s presence within these structures, and monitor RET’s recycling to the cell surface. This method improves on static and less discriminatory methods, providing a tool for analysis of real-time vesicle trafficking that is applicable to many systems.


Membrane protein Internalization Recycling Biotinylation RET 

Supplementary material

10895_2009_548_MOESM1_ESM.doc (42 kb)
Table 1Data used to define theoretical point spread functions for deconvolution. (DOC 42 kb)


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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Department of Pathology and Molecular Medicine, Division of Cancer Biology and Genetics, Cancer Research InstituteQueen’s UniversityKingstonCanada

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