Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes
2015 (English)In: Journal of Visualized Experiments, ISSN 1940-087X, E-ISSN 1940-087X, no 106, e53113- p.Article in journal (Refereed) Epub ahead of printText
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 mu m(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 beta(1-40) (A beta(1-40)) 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.
Place, publisher, year, edition, pages
JOURNAL OF VISUALIZED EXPERIMENTS , 2015. no 106, e53113- p.
Neuroscience; Issue 106; Amyloid beta; Astrocyte size; Confocal microscope; Surface area; Three-dimensional morphometry; Volume
Medical Image Processing
IdentifiersURN: urn:nbn:se:liu:diva-125167DOI: 10.3791/53113ISI: 000368574400013PubMedID: 26709729OAI: oai:DiVA.org:liu-125167DiVA: diva2:903378
Funding Agencies|County Council of Ostergotland, (Sweden); Cellular and Molecular Research Center at Iran University of Medical Sciences (Tehran, Iran)2016-02-152016-02-152016-08-19