title: Workflow for automated quantification of cerebromicrovascular gelatinase activity.
abstract: 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.
authors.affiliation: Research Programs Unit, Molecular Neurology, and Department of Clinical Neurosciences, University of Helsinki, Helsinki, Finland. Electronic address: olli.s.mattila@helsinki.fi.
authors.forename: Olli S
authors.initials: OS
authors.lastname: Mattila
authors.affiliation: Research Programs Unit, Genome-Scale Biology, Institute of Biomedicine, Biochemistry and Developmental Biology, University of Helsinki, Finland.
authors.forename: Ville
authors.initials: V
authors.lastname: Rantanen
authors.affiliation: Research Programs Unit, Molecular Neurology, and Department of Clinical Neurosciences, University of Helsinki, Helsinki, Finland.
authors.forename: Jani
authors.initials: J
authors.lastname: Saksi
authors.affiliation: Department of Neurology, Helsinki University Central Hospital, Finland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Strbian
authors.affiliation: Research Programs Unit, Molecular Neurology, and Department of Clinical Neurosciences, University of Helsinki, Helsinki, Finland.
authors.forename: Tero
authors.initials: T
authors.lastname: Pikkarainen
authors.affiliation: Research Programs Unit, Genome-Scale Biology, Institute of Biomedicine, Biochemistry and Developmental Biology, University of Helsinki, Finland.
authors.forename: Sampsa
authors.initials: S
authors.lastname: Hautaniemi
authors.affiliation: Research Programs Unit, Molecular Neurology, and Department of Clinical Neurosciences, University of Helsinki, Helsinki, Finland; Department of Neurology, Helsinki University Central Hospital, Finland.
authors.forename: Perttu J
authors.initials: PJ
authors.lastname: Lindsberg
mesh_terms: D000818:Animals; D057205:Automation, Laboratory; D001921:Brain / Q000098:blood supply*; D004195:Disease Models, Animal; D018093:Gelatinases / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D020244:Infarction, Middle Cerebral Artery / Q000201:enzymology*; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods*; D055806:Microvessels / Q000201:enzymology*; D017208:Rats, Wistar; D015203:Reproducibility of Results; D057188:Workflow
chemical_list: D018093:Gelatinases
keywords: Automated analysis; Bioinformatics; Experimental stroke; Gelatinases; Image analysis