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Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles

Fluorescence probes have widely been used for detecting and imaging Ca(2+)‐enriched parts of cells but more rarely for quantitative determination of concentrations. In this study we show how this can be achieved by a novel approach using hydrogel particles. In a microfluidic co‐flow arrangement sphe...

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Autores principales: Kronfeld, Klaus‐Peter, Ellinger, Thomas, Köhler, Johann Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8456324/
https://www.ncbi.nlm.nih.gov/pubmed/34584516
http://dx.doi.org/10.1002/elsc.202100023
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author Kronfeld, Klaus‐Peter
Ellinger, Thomas
Köhler, Johann Michael
author_facet Kronfeld, Klaus‐Peter
Ellinger, Thomas
Köhler, Johann Michael
author_sort Kronfeld, Klaus‐Peter
collection PubMed
description Fluorescence probes have widely been used for detecting and imaging Ca(2+)‐enriched parts of cells but more rarely for quantitative determination of concentrations. In this study we show how this can be achieved by a novel approach using hydrogel particles. In a microfluidic co‐flow arrangement spherical droplets were generated from an aqueous solution of acrylamide, N,N'‐methylenebisacrylamide crosslinker and photoinitiator and subsequently photo‐cured in situ yielding gel particles in a sub millimeter range. These particles were separated, dried under reduced pressure and re‐swollen in water containing Rhod‐5N tri potassium salt as calcium ion selective fluorescence probe. After that the particles were dried again and stored for further investigations. Upon exposure of dried particles to calcium chloride solutions they swell and take up Ca(2+)‐ions forming a strong fluorescing complex with Rhod‐5N. Thus, fluorescence intensity increases with calcium ion concentration. Up to ca. 0.50 mM the enhancement effect is strong and then becomes considerably weaker. The intensity‐concentration‐dependence is well described by an equation derived from the equilibrium of the formation of a 1:1 Ca(2+):Rhod‐5N complex. The particles allow for a fast optical determination of Ca(2+)‐concentrations up to 0.50 mM in analyte volumes down to below 10 μL.
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spelling pubmed-84563242021-09-27 Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles Kronfeld, Klaus‐Peter Ellinger, Thomas Köhler, Johann Michael Eng Life Sci Research Articles Fluorescence probes have widely been used for detecting and imaging Ca(2+)‐enriched parts of cells but more rarely for quantitative determination of concentrations. In this study we show how this can be achieved by a novel approach using hydrogel particles. In a microfluidic co‐flow arrangement spherical droplets were generated from an aqueous solution of acrylamide, N,N'‐methylenebisacrylamide crosslinker and photoinitiator and subsequently photo‐cured in situ yielding gel particles in a sub millimeter range. These particles were separated, dried under reduced pressure and re‐swollen in water containing Rhod‐5N tri potassium salt as calcium ion selective fluorescence probe. After that the particles were dried again and stored for further investigations. Upon exposure of dried particles to calcium chloride solutions they swell and take up Ca(2+)‐ions forming a strong fluorescing complex with Rhod‐5N. Thus, fluorescence intensity increases with calcium ion concentration. Up to ca. 0.50 mM the enhancement effect is strong and then becomes considerably weaker. The intensity‐concentration‐dependence is well described by an equation derived from the equilibrium of the formation of a 1:1 Ca(2+):Rhod‐5N complex. The particles allow for a fast optical determination of Ca(2+)‐concentrations up to 0.50 mM in analyte volumes down to below 10 μL. John Wiley and Sons Inc. 2021-06-04 /pmc/articles/PMC8456324/ /pubmed/34584516 http://dx.doi.org/10.1002/elsc.202100023 Text en © 2021 The Authors. Engineering in Life Sciences published by Wiley‐VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Kronfeld, Klaus‐Peter
Ellinger, Thomas
Köhler, Johann Michael
Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles
title Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles
title_full Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles
title_fullStr Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles
title_full_unstemmed Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles
title_short Micro flow photochemical synthesis of Ca‐sensitive fluorescent sensor particles
title_sort micro flow photochemical synthesis of ca‐sensitive fluorescent sensor particles
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8456324/
https://www.ncbi.nlm.nih.gov/pubmed/34584516
http://dx.doi.org/10.1002/elsc.202100023
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