Springer Protocols: Abstract: Visualizing Calcium Signaling in Cells by Digitized Wide-Field and Confocal Fluorescent Microscopy

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

By: Michael Wm. Roe², Jerome F. Fiekers³, Louis H. Philipson², Vytautas P. Bindokas⁴

Abstract

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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.

Affiliation(s): (2) Department of Medicine, University of Chicago, Chicago, IL
(3) Department of Anatomy and Neurobiology, University of Vermont, Burlington, VT
(4) Department of Neurobiology, Pharmacology, and Physiology, University of Chicago, Chicago, IL

Book Title: Cell Imaging Techniques: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 319 | Pub. Date: Nov-01-2005 | Page Range: 37-66 | DOI: 10.1007/978-1-59259-993-6_3

Subject: Imaging/Radiology

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

References

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