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Ratiometric Ca2+ Measurements Using the FlexStation®Scanning Fluorometer

  • Ian C. B. Marshall
  • Izzy Boyfield
  • Shaun McNulty
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 937)

Abstract

The FlexStation ® Scanning Fluorometer is a fluorescence plate reader that can measure intracellular Ca2+ concentration using both single-wavelength and dual-wavelength fluorescent probes. The FlexStation uses a Xenon flashlamp and monochromators for both excitation and emission light to allow the use of a wide range of fluorescent indicators. The system incorporates a fluid transfer system for addition of test compounds from a source plate to the cell plate during data acquisition. Both plates are contained within a temperature-controlled unit that can be controlled accurately between room temperature and 45°C. The FlexStation can be configured to read a range of plate sizes. In this chapter generic methods for assessing intracellular Ca2+ on the FlexStation using ratiometric dyes are described.

Key words

Calcium Flexstation High throughput 

Reference

  1. 1.
    Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Ian C. B. Marshall
    • 1
  • Izzy Boyfield
    • 1
  • Shaun McNulty
    • 1
  1. 1.Neurology & GI Centre of Excellence for Drug Discovery, GlaxoSmithKline Research and Development LimitedHarlowUK

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