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

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), 2024. Published by Science Publishing Group

Keywords

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

References
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    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
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    PY  - 2017
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    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
  • FEMTO-ST Institute, UMR CNRS 6174, Université Bourgogne Franche-Comté, Besan?on, France

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

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

  • Center of Excellence in Molecular Imaging, Chiang Mai University, Chiang Mai, Thailand

  • FEMTO-ST Institute, UMR CNRS 6174, Université Bourgogne Franche-Comté, Besan?on, France; INSERM CIC 1431, Besan?on University Hospital, Besan?on, France

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