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Visualizing Calcium Signaling in Cells by Digitized Wide-Field and Confocal Fluorescent Microscopy

  • Michael Wm. Roe
  • Jerome F. Fiekers
  • Louis H. Philipson
  • Vytautas P. Bindokas
Part of the Methods in Molecular Biology™ book series (MIMB, volume 319)

Abstract

Calcium (Ca2+) is a fundamentally important component of cellular signal transduction. Dynamic changes in the concentration of Ca2+ ([Ca2+]) in the cytoplasm and within organelles are tightly controlled and regulate a diverse array of biological activities, including fertilization, cell division, gene expression, cellular metabolism, protein biosynthesis, secretion, muscle contraction, intercellular communication, and cell death. Measurement of intracellular [Ca2+] is essential to understanding the role of Ca2+ and for defining the underlying regulatory mechanisms in any cellular process. A broad range of synthetic and biosynthetic fluorescent Ca2+ sensors are available that enable the visualization and quantification of subcellular spatio-temporal [Ca2+] gradients. This chapter describes the application of wide-field digitized video fluorescence microfluorometry and confocal microscopy to quantitatively image Ca2+ in cells with high temporal and spatial resolution.

Key Words

Intracellular calcium signal transduction cellular imaging fluorescence confocal microscopy Fura-2 Fluo-3 cameleon pericam biosensor transfection 

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Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Michael Wm. Roe
    • 1
  • Jerome F. Fiekers
    • 2
  • Louis H. Philipson
    • 1
  • Vytautas P. Bindokas
    • 3
  1. 1.Department of MedicineUniversity of ChicagoChicago
  2. 2.Department of Anatomy and NeurobiologyUniversity of VermontBurlington
  3. 3.Department of Neurobiology, Pharmacology, and PhysiologyUniversity of ChicagoChicago

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