Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa.

Schneidereit D, Vass H, Reischl B, Allen RJ, Friedrich O (2016)

Publication Language: English

Publication Status: Published

Publication Type: Journal article

Publication year: 2016

Journal

PLoS ONE Public Library of Science

Book Volume: 11

Pages Range: e0164509

Journal Issue: 10

DOI: 10.1371/journal.pone.0164509

Abstract

The fluorescent Ca2+ sensitive dyes Fura Red (ratiometric) and Fluo-4 (non-ratiometric) are widely utilized for the optical assessment of Ca2+ fluctuations in vitro as well as in situ. The fluorescent behavior of these dyes is strongly depends on temperature, pH, ionic strength and pressure. It is crucial to understand the response of these dyes to pressure when applying calcium imaging technologies in the field of high pressure bioscience. Therefore, we use an optically accessible pressure vessel to pressurize physiological Ca2+-buffered solutions at different fixed concentrations of free Ca2+ (1 nM to 25.6 μM) and a specified dye concentration (12 μM) to pressures of 200 MPa, and record dye fluorescence intensity. Our results show that Fluo-4 fluorescence intensity is reduced by 31% per 100 MPa, the intensity of Fura Red is reduced by 10% per 100 MPa. The mean reaction volume for the dissociation of calcium from the dye molecules [Formula: see text] is determined to -17.8 ml mol-1 for Fluo-4 and -21.3 ml mol-1 for Fura Red. Additionally, a model is presented that is used to correct for pressure-dependent changes in pH and binding affinity of Ca2+ to EGTA, as well as to determine the influence of these changes on dye fluorescence.

Authors with CRIS profile

Dominik Schneidereit Lehrstuhl für Medizinische Biotechnologie Barbara Reischl Lehrstuhl für Medizinische Biotechnologie Oliver Friedrich Lehrstuhl für Medizinische Biotechnologie

Related research project(s)

Skeletal Muscle Function under Pressure: Elucidating Mechanisms of Muscle Ca2+ Signaling Failure and Ultrastructure Disintegration using a novel High-Pressure Multiphoton Microscopy Technology (PiezoGRIN) March 1, 2014 - July 31, 2018

Involved external institutions

University of Edinburgh GB United Kingdom (GB)

How to cite

APA:

Schneidereit, D., Vass, H., Reischl, B., Allen, R.J., & Friedrich, O. (2016). Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa. PLoS ONE, 11(10), e0164509. https://dx.doi.org/10.1371/journal.pone.0164509

MLA:

Schneidereit, Dominik, et al. "Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa." PLoS ONE 11.10 (2016): e0164509.

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