title: Automated Protein Localization of Blood Brain Barrier Vasculature in Brightfield IHC Images.
abstract: In this paper, we present an objective method for localization of proteins in blood brain barrier (BBB) vasculature using standard immunohistochemistry (IHC) techniques and bright-field microscopy. Images from the hippocampal region at the BBB are acquired using bright-field microscopy and subjected to our segmentation pipeline which is designed to automatically identify and segment microvessels containing the protein glucose transporter 1 (GLUT1). Gabor filtering and k-means clustering are employed to isolate potential vascular structures within cryosectioned slabs of the hippocampus, which are subsequently subjected to feature extraction followed by classification via decision forest. The false positive rate (FPR) of microvessel classification is characterized using synthetic and non-synthetic IHC image data for image entropies ranging between 3 and 8 bits. The average FPR for synthetic and non-synthetic IHC image data was found to be 5.48% and 5.04%, respectively.
authors.affiliation: Department of Electrical Engineering, Drexel University, Philadelphia, United States of America.
authors.forename: Rajath E
authors.initials: RE
authors.lastname: Soans
authors.affiliation: Department of Medicine, University of Pennsylvania, Philadelphia, United States of America.
authors.forename: Diane C
authors.initials: DC
authors.lastname: Lim
authors.affiliation: Center for Sleep and Circadian Neurobiology, University of Pennsylvania, Philadelphia, United States of America.
authors.forename: Brendan T
authors.initials: BT
authors.lastname: Keenan
authors.affiliation: Department of Medicine, University of Pennsylvania, Philadelphia, United States of America.
authors.forename: Allan I
authors.initials: AI
authors.lastname: Pack
authors.affiliation: Department of Electrical Engineering, Drexel University, Philadelphia, United States of America.
authors.forename: James A
authors.initials: JA
authors.lastname: Shackleford
mesh_terms: D000465:Algorithms; D000818:Animals; D001331:Automation; D001812:Blood-Brain Barrier / Q000378:metabolism*; D016000:Cluster Analysis; D019277:Entropy; D051272:Glucose Transporter Type 1 / Q000378:metabolism*; D006624:Hippocampus / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008297:Male; D008810:Mice, Inbred C57BL; D055806:Microvessels / Q000378:metabolism*; D021381:Protein Transport; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D051272:Glucose Transporter Type 1
references.citation: Curr Alzheimer Res. 2007 Apr;4(2):191-7
references.pmid: 17430246
references.citation: Drug Metab Dispos. 2012 Jun;40(6):1164-9
references.pmid: 22401960
references.citation: Brain. 2007 Feb;130(Pt 2):521-34
references.pmid: 17124188
references.citation: Neuron. 2008 Jan 24;57(2):178-201
references.pmid: 18215617
references.citation: J Drug Target. 2004;12(9-10):635-41
references.pmid: 15621689
references.citation: Anal Bioanal Chem. 2013 May;405(14):4719-28
references.pmid: 23515611
references.citation: Anal Methods. 2013;5(6):1623-1628
references.pmid: 23544036
references.citation: Brain Res. 2007 Feb 23;1134(1):1-11
references.pmid: 17196184
references.citation: Ann Neurol. 2005 Feb;57(2):176-9
references.pmid: 15668963
references.citation: Am J Physiol Heart Circ Physiol. 2005 Jul;289(1):H212-9
references.pmid: 15708958
references.citation: Nat Biotechnol. 2011 Apr;29(4):341-5
references.pmid: 21423189
references.citation: Prog Neurobiol. 2009 Apr;87(4):212-51
references.pmid: 19395337
references.citation: Anal Chem. 2013 Aug 6;85(15):7191-6
references.pmid: 23829295
references.citation: J Neurochem. 2011 Apr;117(2):333-45
references.pmid: 21291474
references.citation: J Pharm Sci. 2013 Sep;102(9):3343-55
references.pmid: 23650139
references.citation: Nat Biotechnol. 2011 Apr;29(4):325-6
references.pmid: 21478846
references.citation: Biomaterials. 2012 Jan;33(2):704-12
references.pmid: 22019122