Abstract
Total Internal Reflection Fluorescence Microscopy is a powerful application of fluorescence microscopy that allows selective imaging of fluorescent molecules that are either in or close to the plasma membrane of a cell. Thus, it is ideally suited to imaging the trafficking of molecules to and from the plasma membrane. Here we describe its use to study the trafficking and exocytosis of the glucose transporter (GLUT4) in response to insulin stimulation.
Key words
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Burchfield JG, Lopez JA, Mele K, Vallotton P, Hughes WE (2010) Exocytotic vesicle behaviour assessed by total internal reflection fluorescence microscopy. Traffic 11:429–439
Lopez JA et al (2009) Identification of a distal GLUT4 trafficking event controlled by actin polymerization. Mol Biol Cell 20:3918–3929
Ng Y et al (2010) Cluster analysis of insulin action in adipocytes reveals a key role for Akt at the plasma membrane. J Biol Chem 285:2245–2257
Zhao P et al (2009) Variations in the requirement for v-SNAREs in GLUT4 trafficking in adipocytes. J Cell Sci 122:3472–3480
Rasband WS, ImageJ, U.S. National Institutes of Health. (Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997–2010)
Mele K, Burchfield JG, James DE, Vallotton P, Hughes WE (2009) Towards fully automated identification of vesicle-membrane fusion events in TIRF microscopy. Int J Comput Aided Eng Technol 1:502–515
Mele K et al (2009) Automatic identification of fusion events in TIRF microscopy image sequences. In: ICCV09 1st workshop on Video-oriented Object and Event Classification (VOEC’09), Kyoto, Japan, 2009
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Burchfield, J.G., Lopez, J.A., Hughes, W.E. (2012). Using Total Internal Reflection Fluorescence Microscopy (TIRFM) to Visualise Insulin Action. In: Badoer, E. (eds) Visualization Techniques. Neuromethods, vol 70. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-897-9_5
Download citation
DOI: https://doi.org/10.1007/978-1-61779-897-9_5
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-896-2
Online ISBN: 978-1-61779-897-9
eBook Packages: Springer Protocols