Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.

Ca²⁺ release into the cytosol through inositol 1,4,5-trisphosphate receptors (IP₃Rs) plays a relevant role in numerous physiological processes. IP₃R-mediated Ca²⁺ signals involve Ca²⁺-induced Ca²⁺-release (CICR) whereby Ca²⁺ release through one open IP₃R induces the opening of other channels. IP₃Rs...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Estefanía Piegari, Lucía Lopez, Emiliano Perez Ipiña, Silvina Ponce Dawson
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a85807205c9b4c199190155cde01edc1
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a85807205c9b4c199190155cde01edc1
record_format	dspace
spelling	oai:doaj.org-article:a85807205c9b4c199190155cde01edc12021-11-18T08:21:27ZFluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.1932-620310.1371/journal.pone.0095860https://doaj.org/article/a85807205c9b4c199190155cde01edc12014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24776736/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Ca²⁺ release into the cytosol through inositol 1,4,5-trisphosphate receptors (IP₃Rs) plays a relevant role in numerous physiological processes. IP₃R-mediated Ca²⁺ signals involve Ca²⁺-induced Ca²⁺-release (CICR) whereby Ca²⁺ release through one open IP₃R induces the opening of other channels. IP₃Rs are apparently organized in clusters. The signals can remain localized (i.e., Ca²⁺ puffs) if CICR is limited to one cluster or become waves that propagate between clusters. Ca²⁺ puffs are the building blocks of Ca²⁺ waves. Thus, there is great interest in determining puff properties, especially in view of the current controversy on the spatial distribution of activatable IP₃Rs. Ca²⁺ puffs have been observed in intact cells with optical techniques proving that they are intrinsically Ca²⁺ dyes, slow exogenous buffers (e.g., EGTA) to disrupt inter-cluster CICR and UV-photolyzable caged IP3. Single-wavelength dyes increase their fluorescence upon calcium binding producing images that are strongly dependent on their kinetic, transport and photophysical properties. Determining the artifacts that the imaging setting introduces is particularly relevant when trying to analyze the smallest Ca²⁺ signals. In this paper we introduce a method to estimate the expected signal-to-noise ratio of Ca²⁺ imaging experiments that use single-wavelength dyes. The method is based on the Number and rightness technique. It involves the performance of a series of experiments and their subsequent analysis in terms of a fluorescence fluctuation model with which the model parameters are quantified. Using the model, the expected signal-to-noise ratio is then computed. Equivalence classes between different experimental conditions that produce images with similar signal-to-noise ratios can then be established. The method may also be used to estimate the smallest signals that can reliably be observed with each setting.Estefanía PiegariLucía LopezEmiliano Perez IpiñaSilvina Ponce DawsonPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 4, p e95860 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Estefanía Piegari Lucía Lopez Emiliano Perez Ipiña Silvina Ponce Dawson Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.
description	Ca²⁺ release into the cytosol through inositol 1,4,5-trisphosphate receptors (IP₃Rs) plays a relevant role in numerous physiological processes. IP₃R-mediated Ca²⁺ signals involve Ca²⁺-induced Ca²⁺-release (CICR) whereby Ca²⁺ release through one open IP₃R induces the opening of other channels. IP₃Rs are apparently organized in clusters. The signals can remain localized (i.e., Ca²⁺ puffs) if CICR is limited to one cluster or become waves that propagate between clusters. Ca²⁺ puffs are the building blocks of Ca²⁺ waves. Thus, there is great interest in determining puff properties, especially in view of the current controversy on the spatial distribution of activatable IP₃Rs. Ca²⁺ puffs have been observed in intact cells with optical techniques proving that they are intrinsically Ca²⁺ dyes, slow exogenous buffers (e.g., EGTA) to disrupt inter-cluster CICR and UV-photolyzable caged IP3. Single-wavelength dyes increase their fluorescence upon calcium binding producing images that are strongly dependent on their kinetic, transport and photophysical properties. Determining the artifacts that the imaging setting introduces is particularly relevant when trying to analyze the smallest Ca²⁺ signals. In this paper we introduce a method to estimate the expected signal-to-noise ratio of Ca²⁺ imaging experiments that use single-wavelength dyes. The method is based on the Number and rightness technique. It involves the performance of a series of experiments and their subsequent analysis in terms of a fluorescence fluctuation model with which the model parameters are quantified. Using the model, the expected signal-to-noise ratio is then computed. Equivalence classes between different experimental conditions that produce images with similar signal-to-noise ratios can then be established. The method may also be used to estimate the smallest signals that can reliably be observed with each setting.
format	article
author	Estefanía Piegari Lucía Lopez Emiliano Perez Ipiña Silvina Ponce Dawson
author_facet	Estefanía Piegari Lucía Lopez Emiliano Perez Ipiña Silvina Ponce Dawson
author_sort	Estefanía Piegari
title	Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.
title_short	Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.
title_full	Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.
title_fullStr	Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.
title_full_unstemmed	Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.
title_sort	fluorescence fluctuations and equivalence classes of ca²⁺ imaging experiments.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/a85807205c9b4c199190155cde01edc1
work_keys_str_mv	AT estefaniapiegari fluorescencefluctuationsandequivalenceclassesofca2imagingexperiments AT lucialopez fluorescencefluctuationsandequivalenceclassesofca2imagingexperiments AT emilianoperezipina fluorescencefluctuationsandequivalenceclassesofca2imagingexperiments AT silvinaponcedawson fluorescencefluctuationsandequivalenceclassesofca2imagingexperiments
_version_	1718421879135404032

Fluorescence fluctuations and equivalence classes of Ca²⁺ imaging experiments.

Ejemplares similares