CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses

Abstract Intracellular calcium acts as a secondary messenger in a wide variety of crucial biological signaling processes. Advances in fluorescence microscopy and calcium sensitive dyes has led to the routine quantification of calcium responses in non-excitable cells. However, the automatization of g...

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Autores principales:	Angela M. Lee, Huw Colin-York, Marco Fritzsche
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/962b383b6a2c4f46985f169ee6eaa090
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spelling	oai:doaj.org-article:962b383b6a2c4f46985f169ee6eaa0902021-12-02T15:05:37ZCalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses10.1038/s41598-017-05322-z2045-2322https://doaj.org/article/962b383b6a2c4f46985f169ee6eaa0902017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05322-zhttps://doaj.org/toc/2045-2322Abstract Intracellular calcium acts as a secondary messenger in a wide variety of crucial biological signaling processes. Advances in fluorescence microscopy and calcium sensitive dyes has led to the routine quantification of calcium responses in non-excitable cells. However, the automatization of global intracellular calcium analysis at the single-cell level within a large population simultaneously remains challenging. One software, CalQuo (Calcium Quantification), offers some automatic features in calcium analysis. Here, we present an advanced version of the software package: CalQuo 2 . CalQuo 2 analyzes the calcium response in the Fourier-domain, allowing the number of user-defined filtering parameters to be reduced to one and a greater diversity of calcium responses to be recognized, compared to CalQuo that directly interprets the calcium intensity signal. CalQuo 2 differentiates cells that release a single calcium response and those that release oscillatory calcium fluxes. We have demonstrated the use of CalQuo 2 by measuring the calcium response in genetically modified Jurkat T-cells under varying ligand conditions, in which we show that peptide:MHCs and anti-CD3 antibodies trigger a fraction of T cells to release oscillatory calcium fluxes that increase with increasing koff rates. These results show that CalQuo 2 is a robust and user-friendly tool for characterizing global, single cell calcium responses.Angela M. LeeHuw Colin-YorkMarco FritzscheNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Angela M. Lee Huw Colin-York Marco Fritzsche CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses
description	Abstract Intracellular calcium acts as a secondary messenger in a wide variety of crucial biological signaling processes. Advances in fluorescence microscopy and calcium sensitive dyes has led to the routine quantification of calcium responses in non-excitable cells. However, the automatization of global intracellular calcium analysis at the single-cell level within a large population simultaneously remains challenging. One software, CalQuo (Calcium Quantification), offers some automatic features in calcium analysis. Here, we present an advanced version of the software package: CalQuo 2 . CalQuo 2 analyzes the calcium response in the Fourier-domain, allowing the number of user-defined filtering parameters to be reduced to one and a greater diversity of calcium responses to be recognized, compared to CalQuo that directly interprets the calcium intensity signal. CalQuo 2 differentiates cells that release a single calcium response and those that release oscillatory calcium fluxes. We have demonstrated the use of CalQuo 2 by measuring the calcium response in genetically modified Jurkat T-cells under varying ligand conditions, in which we show that peptide:MHCs and anti-CD3 antibodies trigger a fraction of T cells to release oscillatory calcium fluxes that increase with increasing koff rates. These results show that CalQuo 2 is a robust and user-friendly tool for characterizing global, single cell calcium responses.
format	article
author	Angela M. Lee Huw Colin-York Marco Fritzsche
author_facet	Angela M. Lee Huw Colin-York Marco Fritzsche
author_sort	Angela M. Lee
title	CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses
title_short	CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses
title_full	CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses
title_fullStr	CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses
title_full_unstemmed	CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses
title_sort	calquo 2 : automated fourier-space, population-level quantification of global intracellular calcium responses
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/962b383b6a2c4f46985f169ee6eaa090
work_keys_str_mv	AT angelamlee calquo2automatedfourierspacepopulationlevelquantificationofglobalintracellularcalciumresponses AT huwcolinyork calquo2automatedfourierspacepopulationlevelquantificationofglobalintracellularcalciumresponses AT marcofritzsche calquo2automatedfourierspacepopulationlevelquantificationofglobalintracellularcalciumresponses
_version_	1718388782137344000

CalQuo 2 : Automated Fourier-space, population-level quantification of global intracellular calcium responses

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