Dynamics of astrocytes Ca2+ 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 Ca2+ signalling in astrocytes. The created platform, consisting of a spe...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2024.1320672/full |
| _version_ | 1827376313885786112 |
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| author | Rosa Musotto Ulderico Wanderlingh Angela D’Ascola Michela Spatuzza Maria Vincenza Catania Maurizio De Pittà Maurizio De Pittà Maurizio De Pittà Maurizio De Pittà Giovanni Pioggia |
| author_facet | Rosa Musotto Ulderico Wanderlingh Angela D’Ascola Michela Spatuzza Maria Vincenza Catania Maurizio De Pittà Maurizio De Pittà Maurizio De Pittà Maurizio De Pittà Giovanni Pioggia |
| author_sort | Rosa Musotto |
| collection | DOAJ |
| description | 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 Ca2+ 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 Ca2+ ions in astrocytes. To demonstrate the usefulness of the workflow, we applied the methodology to several in vitro 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). |
| first_indexed | 2024-03-08T12:08:04Z |
| format | Article |
| id | doaj.art-c46df195b2e342dfa68a1ce04b754141 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-03-08T12:08:04Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-c46df195b2e342dfa68a1ce04b7541412024-01-23T04:44:33ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2024-01-011210.3389/fcell.2024.13206721320672Dynamics of astrocytes Ca2+ signaling: a low-cost fluorescence customized system for 2D culturesRosa Musotto0Ulderico Wanderlingh1Angela D’Ascola2Michela Spatuzza3Maria Vincenza Catania4Maurizio De Pittà5Maurizio De Pittà6Maurizio De Pittà7Maurizio De Pittà8Giovanni Pioggia9Institute for Biomedical Research and Innovation, National Research Council (IRIB-CNR), Messina, ItalyDepartment of Mathematical and Computer Sciences, Physical Sciences and Earth Sciences, University of Messina, Messina, ItalyDepartment of Clinical and Experimental Medicine, University of Messina, Policlinico Universitario, Messina, ItalyInstitute for Biomedical Research and Innovation, National Research Council (IRIB-CNR), Catania, ItalyInstitute for Biomedical Research and Innovation, National Research Council (IRIB-CNR), Catania, ItalyDivision of Clinical and Computational Neurosciences, Krembil Research Institute, University Health Network, Toronto, ON, CanadaDepartment of Physiology, Temerty Faculty of Medicine, University of Toronto, Toronto, ON, CanadaBasque Center for Applied Mathematics, Bilbao, SpainDepartment of Neurosciences, Faculty of Medicine, The University of the Basque Country (UPV/EHU), Leioa, SpainInstitute for Biomedical Research and Innovation, National Research Council (IRIB-CNR), Messina, ItalyIn 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 Ca2+ 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 Ca2+ ions in astrocytes. To demonstrate the usefulness of the workflow, we applied the methodology to several in vitro 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).https://www.frontiersin.org/articles/10.3389/fcell.2024.1320672/fullastrocytescalcium wavesfluorescencecustomized systemanalysis |
| spellingShingle | Rosa Musotto Ulderico Wanderlingh Angela D’Ascola Michela Spatuzza Maria Vincenza Catania Maurizio De Pittà Maurizio De Pittà Maurizio De Pittà Maurizio De Pittà Giovanni Pioggia Dynamics of astrocytes Ca2+ signaling: a low-cost fluorescence customized system for 2D cultures Frontiers in Cell and Developmental Biology astrocytes calcium waves fluorescence customized system analysis |
| title | Dynamics of astrocytes Ca2+ signaling: a low-cost fluorescence customized system for 2D cultures |
| title_full | Dynamics of astrocytes Ca2+ signaling: a low-cost fluorescence customized system for 2D cultures |
| title_fullStr | Dynamics of astrocytes Ca2+ signaling: a low-cost fluorescence customized system for 2D cultures |
| title_full_unstemmed | Dynamics of astrocytes Ca2+ signaling: a low-cost fluorescence customized system for 2D cultures |
| title_short | Dynamics of astrocytes Ca2+ signaling: a low-cost fluorescence customized system for 2D cultures |
| title_sort | dynamics of astrocytes ca2 signaling a low cost fluorescence customized system for 2d cultures |
| topic | astrocytes calcium waves fluorescence customized system analysis |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2024.1320672/full |
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