Fast fluorescence dynamics in non-ratiometric calcium indicators
M. Gersbach, D. L. Boiko, C. Niclass, C. Petersen, E. Charbon
TL;DR
This paper analyzes the fluorescence decay of Oregon Green BAPTA-1 with high temporal resolution, revealing a triple exponential decay and proposing a new model for precise calcium concentration measurement.
Contribution
It introduces a novel model for accurately measuring free Ca2+ concentration using non-ratiometric indicators based on detailed fluorescence decay analysis.
Findings
Fluorescence decay of OGB-1 is best described by a triple exponential model.
A new measurement model improves accuracy of free Ca2+ concentration.
Provides dissociation constants for non-ratiometric calcium indicators.
Abstract
A fluorescence decay of high-affinity non-ratiometric Ca2+ indicator Oregon Green BAPTA-1 (OGB-1) is analyzed with unprecedented temporal resolution in the two-photon excitation regime. A triple exponential decay is shown to best fit the fluorescence dynamics of OGB-1. We provide a new model for accurate measurements of the free Ca2+ concentration and dissociation constants of non-ratiometric calcium indicators.
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