Abstract
Many commercial organizations currently use the Fluorometric Imaging Plate Reader (FLIPR®: Molecular Devices, Sunnyvale, CA) to conduct highthroughput measurements of intracellular Ca2+ concentration (see Chapter 7 ), taking advantage of its rapid kinetics, reliability, and compatibility for automation. For the majority of industrial applications, the primary limitation of FLIPR (i.e., its requirement for single wavelength fluorescent probes using visible light excitation) is not a significant issue. Indeed, visible light probes offer certain benefits over their ultraviolet (UV)-excited ratiometric counterparts, such as reduced sample autofluorescence and higher absorbance, thereby allowing relatively low concentrations of dye to be used. However, under certain circumstances researchers may prefer to conduct high-throughput experiments with ratiometric dyes, particularly when issues of dye leakage, photobleaching, or signal-to-noise ratio become a concern.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Reference
Grynkiewicz, G., Poenie, M., and Tsien, R. Y. (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260, 3440–3450.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Marshall, I.C.B., Boyfield, I., McNulty, S. (2006). Ratiometric Ca2+ Measurements Using the FlexStation® Scanning Fluorometer. In: Lambert, D.G. (eds) Calcium Signaling Protocols. Methods in Molecular Biology™, vol 312. Humana Press. https://doi.org/10.1385/1-59259-949-4:119
Download citation
DOI: https://doi.org/10.1385/1-59259-949-4:119
Publisher Name: Humana Press
Print ISBN: 978-1-58829-442-5
Online ISBN: 978-1-59259-949-3
eBook Packages: Springer Protocols