{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T04:02:58Z","timestamp":1770523378868,"version":"3.49.0"},"reference-count":39,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2024,6,3]],"date-time":"2024-06-03T00:00:00Z","timestamp":1717372800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Foundation for Science and Technology (FCT)","award":["CEECINST\/00018\/2021"],"award-info":[{"award-number":["CEECINST\/00018\/2021"]}]},{"name":"Foundation for Science and Technology (FCT)","award":["LCF\/PR\/HR21\/52410024"],"award-info":[{"award-number":["LCF\/PR\/HR21\/52410024"]}]},{"name":"Foundation for Science and Technology (FCT)","award":["UIDB\/50026\/2020"],"award-info":[{"award-number":["UIDB\/50026\/2020"]}]},{"name":"Foundation for Science and Technology (FCT)","award":["UIDP\/50026\/2020"],"award-info":[{"award-number":["UIDP\/50026\/2020"]}]},{"name":"Foundation for Science and Technology (FCT)","award":["LA\/P\/0050\/2020"],"award-info":[{"award-number":["LA\/P\/0050\/2020"]}]},{"name":"\u201cla Caixa\u201d Foundation","award":["CEECINST\/00018\/2021"],"award-info":[{"award-number":["CEECINST\/00018\/2021"]}]},{"name":"\u201cla Caixa\u201d Foundation","award":["LCF\/PR\/HR21\/52410024"],"award-info":[{"award-number":["LCF\/PR\/HR21\/52410024"]}]},{"name":"\u201cla Caixa\u201d Foundation","award":["UIDB\/50026\/2020"],"award-info":[{"award-number":["UIDB\/50026\/2020"]}]},{"name":"\u201cla Caixa\u201d Foundation","award":["UIDP\/50026\/2020"],"award-info":[{"award-number":["UIDP\/50026\/2020"]}]},{"name":"\u201cla Caixa\u201d Foundation","award":["LA\/P\/0050\/2020"],"award-info":[{"award-number":["LA\/P\/0050\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Astrocytes are ubiquitous in the brain and spinal cord and display a complex morphology important for the local interactions with neighboring cells, resulting in the modulation of circuit function. Thus, studies focusing on astrocyte physiology in the healthy and diseased brain generally present analyses of astrocytic structure. The labeling method used to visualize the astrocytic structure defines the morphological level to observe and may vary depending on the anatomical sub-regions. The method choice may significantly affect our understanding of their structural diversity. The main goal of this work was to identify a straightforward and efficient protocol for labeling and reconstructing a detailed astrocytic structure to apply and validate in different brain tissue preparations across laboratories. For that, we explored different tissue processing protocols before GFAP labeling to determine the most effective method for reconstructing astrocytic backbones in the mouse hippocampus. Our results show that the reconstruction of astrocytic structure in vibratome sections labeled by free-floating immunofluorescence protocol provides a more practical method to achieve a higher level of detail and arbor complexity in astrocyte backbone reconstruction. Free-floating immunofluorescence labeling is the most reliable method for obtaining better antibody penetration and more detailed astrocyte structure. Finally, we also show that introducing an antigen retrieval step appears useful for visualizing more complete structural details.<\/jats:p>","DOI":"10.3390\/cells13110969","type":"journal-article","created":{"date-parts":[[2024,6,3]],"date-time":"2024-06-03T12:29:47Z","timestamp":1717417787000},"page":"969","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Assessing Different Histological Preparations for Reconstruction of Astrocyte Tridimensional Structure"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6394-8734","authenticated-orcid":false,"given":"Sara","family":"Barsanti","sequence":"first","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Jo\u00e3o Filipe","family":"Viana","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Alexandra","family":"Veiga","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Jo\u00e3o Lu\u00eds","family":"Machado","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Daniela Sofia","family":"Abreu","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Jos\u00e9 Duarte","family":"Dias","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3037-9635","authenticated-orcid":false,"given":"Susana","family":"Monteiro","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1061-1716","authenticated-orcid":false,"given":"Nuno A.","family":"Silva","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7724-0446","authenticated-orcid":false,"given":"Lu\u00edsa","family":"Pinto","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Jo\u00e3o Filipe","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal"},{"name":"ICVS\/3B\u2019s-PT Government Associate Laboratory, 4710-057 Braga\/Guimar\u00e3es, Portugal"},{"name":"IPCA-EST-2Ai, Applied Artificial Intelligence Laboratory, Polytechnic Institute of C\u00e1vado and Ave, Campus of IPCA, 4750-810 Barcelos, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2024,6,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1016\/j.neuron.2014.02.007","article-title":"Gliotransmitters Travel in Time and Space","volume":"81","author":"Araque","year":"2014","journal-title":"Neuron"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/j.tins.2015.07.006","article-title":"Do Stars Govern Our Actions? 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