Automated Measurement of Blood Vessels in Tissues from Microscopy Images

Neil J. Kelly1, Nadine Dandachi1, Dmitry A. Goncharov2, Andressa Z. Pena2, Josiah E. Radder1, Alyssa D. Gregory1, Yen‐Chun Lai2, Adriana S. Leme1, Mark T. Gladwin2, Elena A. Goncharova2, Claudette M. St. Croix3, Steven D. Shapiro1

1 Department of Medicine, University of Pittsburgh and University of Pittsburgh Medical Center, Pittsburgh, 2 Vascular Medicine Institute, University of Pittsburgh and University of Pittsburgh Medical Center, Pittsburgh, 3 Center for Biologic Imaging, University of Pittsburgh and University of Pittsburgh Medical Center, Pittsburgh
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 12.44
DOI:  10.1002/cpcy.10
Online Posting Date:  October, 2016
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The quantification of tunica media thickness in histological cross sections is a ubiquitous exercise in cardiopulmonary research, yet the methods for quantifying medial wall thickness have never been rigorously examined with modern image analysis tools. As a result, inaccurate and cumbersome manual measurements of discrete wall regions along the vessel periphery have become common practice for wall thickness quantification. The aim of this study is to introduce, validate, and facilitate the use of an improved method for medial wall thickness quantification. We describe a novel method of wall thickness calculation based on image skeletonization and compare its results to those of common techniques. Using both theoretical and empirical approaches, we demonstrate the accuracy and superiority of the skeleton‐based method for measuring wall thickness while discussing its interpretation and limitations. Finally, we present a new freely available software tool, the VMI Calculator, to facilitate wall thickness measurements using our novel method. © 2016 by John Wiley & Sons, Inc.

Keywords: morphometry; pulmonary hypertension; wall thickness

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Table of Contents

  • Commentary
  • Literature Cited
  • Figures
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Basic Protocol 1:

  • A computer running Windows (version 7.0 or greater) or Macintosh (version OSX or greater). An Internet connection is required to download the software but is not required for its use. The software can be run through an existing installation of MATLAB (version R2012a or greater) or by installing the software prepackaged with the MATLAB Runtime; a MATLAB license is not required for the latter installation. The minimum software requirements are 500 MB of hard disk space and 4 GB of memory.
  • Image files in RGB format (.tif or .jpg are recommended) showing vessel cross sections to be analyzed. Immunofluorescent images are recommended to provide maximum contrast for automated identification of vessel areas; other staining techniques can be used but may require manual identification of vessel areas. At least one image file in a set should have a scale bar for conversion of pixel widths to microns.
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Literature Cited

Literature Cited
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