title: Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes.
abstract: As glial cells in the brain, astrocytes have diverse functional roles in the central nervous system. In the presence of harmful stimuli, astrocytes modify their functional and structural properties, a condition called reactive astrogliosis. Here, a protocol for assessment of the morphological properties of astrocytes is presented. This protocol includes quantification of 12 different parameters i.e. the surface area and volume of the tissue covered by an astrocyte (astrocyte territory), the entire astrocyte including branches, cell body, and nucleus, as well as total length and number of branches, the intensity of fluorescence immunoreactivity of antibodies used for astrocyte detection, and astrocyte density (number/1,000 µm(2)). For this purpose three-dimensional (3D) confocal microscopic images were created, and 3D image analysis software such as Volocity 6.3 was used for measurements. Rat brain tissue exposed to amyloid beta1-40 (Aβ1-40) with or without a therapeutic intervention was used to present the method. This protocol can also be used for 3D morphometric analysis of other cells from either in vivo or in vitro conditions.
authors.affiliation: Department of Clinical and Experimental Medicine, Linköping University; bagheri_ph@yahoo.com.
authors.forename: Maryam
authors.initials: M
authors.lastname: Bagheri
authors.affiliation: Department of Clinical and Experimental Medicine, Linköping University.
authors.forename: Arjang
authors.initials: A
authors.lastname: Rezakhani
authors.affiliation: Neurophysiology Research Center, Shahed University.
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Roghani
authors.affiliation: Cellular and Molecular Research Center, Iran University of Medical Sciences; School of Advanced Technologies in Medicine, Iran University of Medical Sciences.
authors.forename: Mohammad T
authors.initials: MT
authors.lastname: Joghataei
authors.affiliation: Department of Clinical and Experimental Medicine, Linköping University.
authors.forename: Simin
authors.initials: S
authors.lastname: Mohseni
mesh_terms: D016229:Amyloid beta-Peptides / Q000494:pharmacology; D000818:Animals; D001253:Astrocytes / Q000166:cytology*; D001921:Brain / Q000166:cytology / Q000187:drug effects; D021621:Imaging, Three-Dimensional / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods*; D010446:Peptide Fragments / Q000494:pharmacology; D051381:Rats
chemical_list: D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; C078592:amyloid beta-protein (1-40)