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Using Total Internal Reflection Fluorescence Microscopy (TIRFM) to Visualise Insulin Action

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Visualization Techniques

Part of the book series: Neuromethods ((NM,volume 70))

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.

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References

  1. 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

    Article  PubMed  CAS  Google Scholar 

  2. Lopez JA et al (2009) Identification of a distal GLUT4 trafficking event controlled by actin polymerization. Mol Biol Cell 20:3918–3929

    Article  PubMed  CAS  Google Scholar 

  3. 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

    Article  PubMed  CAS  Google Scholar 

  4. Zhao P et al (2009) Variations in the requirement for v-SNAREs in GLUT4 trafficking in adipocytes. J Cell Sci 122:3472–3480

    Article  PubMed  CAS  Google Scholar 

  5. Rasband WS, ImageJ, U.S. National Institutes of Health. (Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997–2010)

  6. 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

    Article  Google Scholar 

  7. 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

    Google Scholar 

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Correspondence to James G. Burchfield .

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© 2012 Springer Science+Business Media, LLC

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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

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  • DOI: https://doi.org/10.1007/978-1-61779-897-9_5

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-896-2

  • Online ISBN: 978-1-61779-897-9

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