دورية أكاديمية
أعرض في EDS

Assessing Different Histological Preparations for Reconstruction of Astrocyte Tridimensional Structure.

التفاصيل البيبلوغرافية
العنوان: Assessing Different Histological Preparations for Reconstruction of Astrocyte Tridimensional Structure.
المؤلفون: Barsanti S; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Viana JF; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Veiga A; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Machado JL; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Abreu DS; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Dias JD; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Monteiro S; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Silva NA; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Pinto L; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal., Oliveira JF; Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 4710-057 Braga, Portugal.; ICVS/3B's-PT Government Associate Laboratory, 4710-057 Braga/Guimarães, Portugal.; IPCA-EST-2Ai, Applied Artificial Intelligence Laboratory, Polytechnic Institute of Cávado and Ave, Campus of IPCA, 4750-810 Barcelos, Portugal.
المصدر: Cells [Cells] 2024 Jun 03; Vol. 13 (11). Date of Electronic Publication: 2024 Jun 03.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: MDPI Country of Publication: Switzerland NLM ID: 101600052 Publication Model: Electronic Cited Medium: Internet ISSN: 2073-4409 (Electronic) Linking ISSN: 20734409 NLM ISO Abbreviation: Cells Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Basel, Switzerland : MDPI
مواضيع طبية MeSH: Astrocytes*/metabolism , Astrocytes*/cytology, Animals ; Mice ; Hippocampus/cytology ; Glial Fibrillary Acidic Protein/metabolism ; Mice, Inbred C57BL ; Male ; Staining and Labeling/methods
مستخلص: 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.
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معلومات مُعتمدة: LCF/PR/HR21/52410024 "la Caixa" Foundation
فهرسة مساهمة: Keywords: antibody penetration; astrocyte; immunofluorescence labeling; tridimensional reconstruction
المشرفين على المادة: 0 (Glial Fibrillary Acidic Protein)
تواريخ الأحداث: Date Created: 20240619 Date Completed: 20240619 Latest Revision: 20240620
رمز التحديث: 20240620
مُعرف محوري في PubMed: PMC11171983
DOI: 10.3390/cells13110969
PMID: 38891101
قاعدة البيانات: MEDLINE
الوصف
تدمد:2073-4409
DOI:10.3390/cells13110969