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Bioluminescence Imaging of Calcium Oscillations Inside Intracellular Organelles

  • Carlos Villalobos
  • María Teresa Alonso
  • Javier García-Sancho
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 574)

Abstract

Ca2+ oscillations inside intracellular organelles are important for regulation of functions such as gene expression at the nucleus, respiration at mitochondria or protein processing at the endoplasmic reticulum. Targeted aequorins are excellent calcium probes for subcellular analysis, but single-cell imaging has proven difficult because of low light yield. Here we describe a procedure that combines virus-based expression of targeted aequorins with photon-counting imaging. This methodology allows real-time resolution of changes of cytosolic, mitochondrial or nuclear Ca2+ signals at the single-cell level.

Key words

Bioluminescence imaging aequorin herpes simplex virus calcium oscillations mitochondria nucleus endoplasmic reticulum anterior pituitary pancreatic islets of Langerhans β cells 

Notes

Acknowledgements

Financial support from the Spanish Ministerio de Educación y Ciencia (grant BFU-2006-60157) is gratefully acknowledged.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Carlos Villalobos
    • 1
  • María Teresa Alonso
    • 1
  • Javier García-Sancho
    • 1
  1. 1.Instituto de Biología y Genética Molecular (IBGM)Universidad de Valladolid and Consejo Superior de Investigaciones Científicas (CSIC)ValladolidSpain

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