Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer

Free zinc is involved in signal transduction within mammalian cells, acting as a second messenger. Gold standard for its analysis is currently the use of metal-responsive fluorescent probes. The present study elucidates the impact of instrumentation used for measuring the resulting fluorescence. The...

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Detalles Bibliográficos
Autores principales: Alker, Wiebke, Haase, Hajo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8055576/
https://www.ncbi.nlm.nih.gov/pubmed/32865725
http://dx.doi.org/10.1007/s12011-020-02355-w
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author Alker, Wiebke
Haase, Hajo
author_facet Alker, Wiebke
Haase, Hajo
author_sort Alker, Wiebke
collection PubMed
description Free zinc is involved in signal transduction within mammalian cells, acting as a second messenger. Gold standard for its analysis is currently the use of metal-responsive fluorescent probes. The present study elucidates the impact of instrumentation used for measuring the resulting fluorescence. The free zinc concentration of THP-1 cells loaded with the fluorescent probes Zinpyr-1 (ZP1) or Fluozin-3 AM (FZ3) was determined using a microplate reader (MPR) and a flow cytometer (FC). Depending on the instrumentation, either low nanomolar (MPR) or picomolar (FC) concentrations of free zinc were observed. The concentrations measured from identical samples by MPR were about 40 (ZP1) or 165 (FZ3) times higher compared with FC. These results demonstrate that the choice of instrumentation has a fundamental impact on the determination of intracellular free zinc concentrations by low molecular weight fluorescent probes.
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spelling pubmed-80555762021-05-05 Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer Alker, Wiebke Haase, Hajo Biol Trace Elem Res Article Free zinc is involved in signal transduction within mammalian cells, acting as a second messenger. Gold standard for its analysis is currently the use of metal-responsive fluorescent probes. The present study elucidates the impact of instrumentation used for measuring the resulting fluorescence. The free zinc concentration of THP-1 cells loaded with the fluorescent probes Zinpyr-1 (ZP1) or Fluozin-3 AM (FZ3) was determined using a microplate reader (MPR) and a flow cytometer (FC). Depending on the instrumentation, either low nanomolar (MPR) or picomolar (FC) concentrations of free zinc were observed. The concentrations measured from identical samples by MPR were about 40 (ZP1) or 165 (FZ3) times higher compared with FC. These results demonstrate that the choice of instrumentation has a fundamental impact on the determination of intracellular free zinc concentrations by low molecular weight fluorescent probes. Springer US 2020-08-31 2021 /pmc/articles/PMC8055576/ /pubmed/32865725 http://dx.doi.org/10.1007/s12011-020-02355-w Text en © The Author(s) 2020 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Alker, Wiebke
Haase, Hajo
Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer
title Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer
title_full Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer
title_fullStr Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer
title_full_unstemmed Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer
title_short Comparison of Free Zinc Levels Determined by Fluorescent Probes in THP1 Cells Using Microplate Reader and Flow Cytometer
title_sort comparison of free zinc levels determined by fluorescent probes in thp1 cells using microplate reader and flow cytometer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8055576/
https://www.ncbi.nlm.nih.gov/pubmed/32865725
http://dx.doi.org/10.1007/s12011-020-02355-w
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