Dynamics of astrocytes Ca 2+ signaling: a low-cost fluorescence customized system for 2D cultures

In an effort to help reduce the costs of fluorescence microscopy and expand the use of this valuable technique, we developed a low-cost platform capable of visualising and analysing the spatio-temporal dynamics of intracellular Ca signalling in astrocytes. The created platform, consisting of a speci...

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Published inFrontiers in cell and developmental biology Vol. 12; p. 1320672
Main Authors Musotto, Rosa, Wanderlingh, Ulderico, D'Ascola, Angela, Spatuzza, Michela, Catania, Maria Vincenza, De Pittà, Maurizio, Pioggia, Giovanni
Format Journal Article
LanguageEnglish
Published Switzerland 2024
Subjects
calcium waves
fluorescence
customized system
astrocytes
analysis
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Summary:In an effort to help reduce the costs of fluorescence microscopy and expand the use of this valuable technique, we developed a low-cost platform capable of visualising and analysing the spatio-temporal dynamics of intracellular Ca signalling in astrocytes. The created platform, consisting of a specially adapted fluorescence microscope and a data analysis procedure performed with Imagej Fiji software and custom scripts, allowed us to detect relative changes of intracellular Ca ions in astrocytes. To demonstrate the usefulness of the workflow, we applied the methodology to several astrocyte preparations, specifically immortalised human astrocyte cells and wild-type mouse cells. To demonstrate the reliability of the procedure, analyses were conducted by stimulating astrocyte activity with the agonist dihydroxyphenylglycine (DHPG), alone or in the presence of the antagonist 2-methyl-6-phenylethyl-pyridine (MPEP).
ISSN:2296-634X
2296-634X