Protocol

Measurement of Calibration Constants for Quantitative Calcium Fluorimetry

  1. Erwin Neher
Adapted from Imaging: A Laboratory Manual (ed. Yuste). CSHL Press, Cold Spring Harbor, NY, USA, 2011.

Abstract

A careful calibration of the fluorescence setup is essential when using Fura-2 for quantitative fluorimetry. The calibration procedure described here is used for experiments in which the indicator dye is introduced into the cell through a patch-clamp pipette. The fluorescence ratio is determined in whole-cell recordings for three different calibration parameters that reflect different [Ca2+]: very high (R1), intermediate (Keff), and zero (R0).

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  1. doi:10.1101/pdb.prot078212 Cold Spring Harb Protoc 2013: pdb.prot078212- © 2013 Cold Spring Harbor Laboratory Press
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