Workflow for automated quantification of cerebromicrovascular gelatinase activity

The gelatinase enzymes, matrix metalloproteinases -2 and -9, are central mediators of blood–brain barrier disruption, actively studied in experimental models of neurological disease. Staining with in situ zymography (ISZ) allows visualization of gelatinase activity directly in brain tissue sections....

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Bibliographic Details
Published in:Microvascular research 2015-01, Vol.97, p.19-24
Main Authors: Mattila, Olli S., Rantanen, Ville, Saksi, Jani, Strbian, Daniel, Pikkarainen, Tero, Hautaniemi, Sampsa, Lindsberg, Perttu J.
Format: Article
Language:English
Subjects:
Animals
Automated analysis
Automation, Laboratory
Bioinformatics
Brain - blood supply
Disease Models, Animal
Experimental stroke
Gelatinases
Gelatinases - metabolism
Image analysis
Image Processing, Computer-Assisted - methods
Immunohistochemistry - methods
Infarction, Middle Cerebral Artery - enzymology
Male
Microscopy, Fluorescence - methods
Microvessels - enzymology
Rats, Wistar
Reproducibility of Results
Workflow
Citations: Items that this one cites
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Summary:The gelatinase enzymes, matrix metalloproteinases -2 and -9, are central mediators of blood–brain barrier disruption, actively studied in experimental models of neurological disease. Staining with in situ zymography (ISZ) allows visualization of gelatinase activity directly in brain tissue sections. However, quantifying microvascular gelatinase activity from ISZ-images is challenging and time consuming, as surrounding cell types often show significant confounding activity. We describe validation and performance of a workflow for automated image analysis of cerebromicrovascular gelatinase activity, now released for open-access use. In comparison to manual analysis, the automated workflow showed superior accuracy, was faster to execute and allows for more detailed analysis of heterogeneity in the microvasculature. We further suggest recommendations for quantifying and reporting this type of activity in experimental studies, focusing on ischemic stroke. •In situ zymography can be used to show gelatinase activity of the microvasculature.•Automated image analysis outperforms manual counting of ISZ positive vessels.•We provide a downloadable analysis workflow to improve methodology of quantifying microvascular ISZ activity.•The workflow enables consistent measurements and reporting of intensity and distribution of ISZ activity.
ISSN:0026-2862
1095-9319
DOI:10.1016/j.mvr.2014.08.009