Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing

Rutjapan Kateklum; Bernard Gauthier-Manuel; Christian Pieralli; Samlee Mankhetkorn; Bruno Wacogne

doi:doi:10.11648/j.ijpp.20170102.11

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Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing

Rutjapan Kateklum, Bernard Gauthier-Manuel, Christian Pieralli, Samlee Mankhetkorn, Bruno Wacogne

Published in International Journal of Photochemistry and Photobiology (Volume 1, Issue 2)

Received: 23 February 2017 Accepted: 21 March 2017 Published: 14 April 2017

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Abstract

A simple and easy to implement numerical method is proposed in order to considerably simplify the experimental calibration procedure of C-SNARF-1 indicator used for ratiometric pH sensing. Usually, calibration is based on the measurement of fluorescence spectra using perfectly calibrated equipment at extreme pH values. The calibration solutions must be extremely well controlled in terms of indicator concentration and path length. Also, the optical equipment used must be well controlled and excitation energy as well as fluorescence collection efficiency must be perfectly constant over the whole calibration procedure. The method we propose is based on the fact that the emission fluorescence energy does not only depend on pH but also on the excitation wavelength. In this paper, we propose a model describing the evolution of the emitted energy as a function of pH and excitation wavelength. We show that the emitted energy evolves linearly with pH and we express this linear evolution as a function of the excitation wavelength. We also show the evolution of the isosbestic (or isoemitting) point as a function of the excitation wavelength. Knowing the linear dependence of the emitted energy as a function of excitation wavelength allows post-processing calibration spectra obtained with basic optical equipment where the excitation energy, fluorescence collection efficiency, indicator concentration and path length can vary over the calibration session. Because the calibration procedure becomes independent of the above mentioned parameters, the post-processing we propose considerably simplify indicators calibration. This method can easily be transposed, not only to other ratiometric pH indicators, but also to ion sensing fluorescent indicators exhibiting dual emission peaks.

Published in	International Journal of Photochemistry and Photobiology (Volume 1, Issue 2)
DOI	10.11648/j.ijpp.20170102.11
Page(s)	36-43
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2017. Published by Science Publishing Group

Keywords

Fluorescence pH Sensing, Ratiometric Measurements, Calibration, C-SNARF-1

References

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APA Style

Rutjapan Kateklum, Bernard Gauthier-Manuel, Christian Pieralli, Samlee Mankhetkorn, Bruno Wacogne. (2017). Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing. International Journal of Photochemistry and Photobiology, 1(2), 36-43. https://doi.org/10.11648/j.ijpp.20170102.11

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ACS Style

Rutjapan Kateklum; Bernard Gauthier-Manuel; Christian Pieralli; Samlee Mankhetkorn; Bruno Wacogne. Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing. Int. J. Photochem. Photobiol. 2017, 1(2), 36-43. doi: 10.11648/j.ijpp.20170102.11

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AMA Style

Rutjapan Kateklum, Bernard Gauthier-Manuel, Christian Pieralli, Samlee Mankhetkorn, Bruno Wacogne. Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing. Int J Photochem Photobiol. 2017;1(2):36-43. doi: 10.11648/j.ijpp.20170102.11

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@article{10.11648/j.ijpp.20170102.11,
  author = {Rutjapan Kateklum and Bernard Gauthier-Manuel and Christian Pieralli and Samlee Mankhetkorn and Bruno Wacogne},
  title = {Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing},
  journal = {International Journal of Photochemistry and Photobiology},
  volume = {1},
  number = {2},
  pages = {36-43},
  doi = {10.11648/j.ijpp.20170102.11},
  url = {https://doi.org/10.11648/j.ijpp.20170102.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijpp.20170102.11},
  abstract = {A simple and easy to implement numerical method is proposed in order to considerably simplify the experimental calibration procedure of C-SNARF-1 indicator used for ratiometric pH sensing. Usually, calibration is based on the measurement of fluorescence spectra using perfectly calibrated equipment at extreme pH values. The calibration solutions must be extremely well controlled in terms of indicator concentration and path length. Also, the optical equipment used must be well controlled and excitation energy as well as fluorescence collection efficiency must be perfectly constant over the whole calibration procedure. The method we propose is based on the fact that the emission fluorescence energy does not only depend on pH but also on the excitation wavelength. In this paper, we propose a model describing the evolution of the emitted energy as a function of pH and excitation wavelength. We show that the emitted energy evolves linearly with pH and we express this linear evolution as a function of the excitation wavelength. We also show the evolution of the isosbestic (or isoemitting) point as a function of the excitation wavelength. Knowing the linear dependence of the emitted energy as a function of excitation wavelength allows post-processing calibration spectra obtained with basic optical equipment where the excitation energy, fluorescence collection efficiency, indicator concentration and path length can vary over the calibration session. Because the calibration procedure becomes independent of the above mentioned parameters, the post-processing we propose considerably simplify indicators calibration. This method can easily be transposed, not only to other ratiometric pH indicators, but also to ion sensing fluorescent indicators exhibiting dual emission peaks.},
 year = {2017}
}

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TY  - JOUR
T1  - Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing
AU  - Rutjapan Kateklum
AU  - Bernard Gauthier-Manuel
AU  - Christian Pieralli
AU  - Samlee Mankhetkorn
AU  - Bruno Wacogne
Y1  - 2017/04/14
PY  - 2017
N1  - https://doi.org/10.11648/j.ijpp.20170102.11
DO  - 10.11648/j.ijpp.20170102.11
T2  - International Journal of Photochemistry and Photobiology
JF  - International Journal of Photochemistry and Photobiology
JO  - International Journal of Photochemistry and Photobiology
SP  - 36
EP  - 43
PB  - Science Publishing Group
SN  - 2640-429X
UR  - https://doi.org/10.11648/j.ijpp.20170102.11
AB  - A simple and easy to implement numerical method is proposed in order to considerably simplify the experimental calibration procedure of C-SNARF-1 indicator used for ratiometric pH sensing. Usually, calibration is based on the measurement of fluorescence spectra using perfectly calibrated equipment at extreme pH values. The calibration solutions must be extremely well controlled in terms of indicator concentration and path length. Also, the optical equipment used must be well controlled and excitation energy as well as fluorescence collection efficiency must be perfectly constant over the whole calibration procedure. The method we propose is based on the fact that the emission fluorescence energy does not only depend on pH but also on the excitation wavelength. In this paper, we propose a model describing the evolution of the emitted energy as a function of pH and excitation wavelength. We show that the emitted energy evolves linearly with pH and we express this linear evolution as a function of the excitation wavelength. We also show the evolution of the isosbestic (or isoemitting) point as a function of the excitation wavelength. Knowing the linear dependence of the emitted energy as a function of excitation wavelength allows post-processing calibration spectra obtained with basic optical equipment where the excitation energy, fluorescence collection efficiency, indicator concentration and path length can vary over the calibration session. Because the calibration procedure becomes independent of the above mentioned parameters, the post-processing we propose considerably simplify indicators calibration. This method can easily be transposed, not only to other ratiometric pH indicators, but also to ion sensing fluorescent indicators exhibiting dual emission peaks.
VL  - 1
IS  - 2
ER  -

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Author Information

Rutjapan Kateklum

FEMTO-ST Institute, UMR CNRS 6174, Université Bourgogne Franche-Comté, Besan?on, France
Bernard Gauthier-Manuel

FEMTO-ST Institute, UMR CNRS 6174, Université Bourgogne Franche-Comté, Besan?on, France
Christian Pieralli

FEMTO-ST Institute, UMR CNRS 6174, Université Bourgogne Franche-Comté, Besan?on, France
Samlee Mankhetkorn

Center of Excellence in Molecular Imaging, Chiang Mai University, Chiang Mai, Thailand
Bruno Wacogne

FEMTO-ST Institute, UMR CNRS 6174, Université Bourgogne Franche-Comté, Besan?on, France

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APA Style

Rutjapan Kateklum, Bernard Gauthier-Manuel, Christian Pieralli, Samlee Mankhetkorn, Bruno Wacogne. (2017). Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing. International Journal of Photochemistry and Photobiology, 1(2), 36-43. https://doi.org/10.11648/j.ijpp.20170102.11

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ACS Style

Rutjapan Kateklum; Bernard Gauthier-Manuel; Christian Pieralli; Samlee Mankhetkorn; Bruno Wacogne. Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing. Int. J. Photochem. Photobiol. 2017, 1(2), 36-43. doi: 10.11648/j.ijpp.20170102.11

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AMA Style

Rutjapan Kateklum, Bernard Gauthier-Manuel, Christian Pieralli, Samlee Mankhetkorn, Bruno Wacogne. Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing. Int J Photochem Photobiol. 2017;1(2):36-43. doi: 10.11648/j.ijpp.20170102.11

Copy | Download

@article{10.11648/j.ijpp.20170102.11,
  author = {Rutjapan Kateklum and Bernard Gauthier-Manuel and Christian Pieralli and Samlee Mankhetkorn and Bruno Wacogne},
  title = {Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing},
  journal = {International Journal of Photochemistry and Photobiology},
  volume = {1},
  number = {2},
  pages = {36-43},
  doi = {10.11648/j.ijpp.20170102.11},
  url = {https://doi.org/10.11648/j.ijpp.20170102.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijpp.20170102.11},
  abstract = {A simple and easy to implement numerical method is proposed in order to considerably simplify the experimental calibration procedure of C-SNARF-1 indicator used for ratiometric pH sensing. Usually, calibration is based on the measurement of fluorescence spectra using perfectly calibrated equipment at extreme pH values. The calibration solutions must be extremely well controlled in terms of indicator concentration and path length. Also, the optical equipment used must be well controlled and excitation energy as well as fluorescence collection efficiency must be perfectly constant over the whole calibration procedure. The method we propose is based on the fact that the emission fluorescence energy does not only depend on pH but also on the excitation wavelength. In this paper, we propose a model describing the evolution of the emitted energy as a function of pH and excitation wavelength. We show that the emitted energy evolves linearly with pH and we express this linear evolution as a function of the excitation wavelength. We also show the evolution of the isosbestic (or isoemitting) point as a function of the excitation wavelength. Knowing the linear dependence of the emitted energy as a function of excitation wavelength allows post-processing calibration spectra obtained with basic optical equipment where the excitation energy, fluorescence collection efficiency, indicator concentration and path length can vary over the calibration session. Because the calibration procedure becomes independent of the above mentioned parameters, the post-processing we propose considerably simplify indicators calibration. This method can easily be transposed, not only to other ratiometric pH indicators, but also to ion sensing fluorescent indicators exhibiting dual emission peaks.},
 year = {2017}
}

Copy | Download

TY  - JOUR
T1  - Simplifying Ratiometric C-SNARF-1 pH Calibration Procedures with a Simple Post-Processing
AU  - Rutjapan Kateklum
AU  - Bernard Gauthier-Manuel
AU  - Christian Pieralli
AU  - Samlee Mankhetkorn
AU  - Bruno Wacogne
Y1  - 2017/04/14
PY  - 2017
N1  - https://doi.org/10.11648/j.ijpp.20170102.11
DO  - 10.11648/j.ijpp.20170102.11
T2  - International Journal of Photochemistry and Photobiology
JF  - International Journal of Photochemistry and Photobiology
JO  - International Journal of Photochemistry and Photobiology
SP  - 36
EP  - 43
PB  - Science Publishing Group
SN  - 2640-429X
UR  - https://doi.org/10.11648/j.ijpp.20170102.11
AB  - A simple and easy to implement numerical method is proposed in order to considerably simplify the experimental calibration procedure of C-SNARF-1 indicator used for ratiometric pH sensing. Usually, calibration is based on the measurement of fluorescence spectra using perfectly calibrated equipment at extreme pH values. The calibration solutions must be extremely well controlled in terms of indicator concentration and path length. Also, the optical equipment used must be well controlled and excitation energy as well as fluorescence collection efficiency must be perfectly constant over the whole calibration procedure. The method we propose is based on the fact that the emission fluorescence energy does not only depend on pH but also on the excitation wavelength. In this paper, we propose a model describing the evolution of the emitted energy as a function of pH and excitation wavelength. We show that the emitted energy evolves linearly with pH and we express this linear evolution as a function of the excitation wavelength. We also show the evolution of the isosbestic (or isoemitting) point as a function of the excitation wavelength. Knowing the linear dependence of the emitted energy as a function of excitation wavelength allows post-processing calibration spectra obtained with basic optical equipment where the excitation energy, fluorescence collection efficiency, indicator concentration and path length can vary over the calibration session. Because the calibration procedure becomes independent of the above mentioned parameters, the post-processing we propose considerably simplify indicators calibration. This method can easily be transposed, not only to other ratiometric pH indicators, but also to ion sensing fluorescent indicators exhibiting dual emission peaks.
VL  - 1
IS  - 2
ER  -

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