title: Automated muscle fiber type population analysis with ImageJ of whole rat muscles using rapid myosin heavy chain immunohistochemistry.
abstract: Skeletal muscle consists of different fiber types which adapt to exercise, aging, disease, or trauma. Here we present a protocol for fast staining, automatic acquisition, and quantification of fiber populations with ImageJ.
  
  
  
  Biceps and lumbrical muscles were harvested from Sprague-Dawley rats. Quadruple immunohistochemical staining was performed on single sections using antibodies against myosin heavy chains and secondary fluorescent antibodies. Slides were scanned automatically with a slide scanner. Manual and automatic analyses were performed and compared statistically.
  
  
  
  The protocol provided rapid and reliable staining for automated image acquisition. Analyses between manual and automatic data indicated Pearson correlation coefficients for biceps of 0.645-0.841 and 0.564-0.673 for lumbrical muscles. Relative fiber populations were accurate to a degree of ± 4%.
  
  
  
  This protocol provides a reliable tool for quantification of muscle fiber populations. Using freely available software, it decreases the required time to analyze whole muscle sections. Muscle Nerve 54: 292-299, 2016.
authors.affiliation: CD Laboratory for the Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Spitalgasse 23, A-1090, Austria.
authors.forename: Konstantin D
authors.identifier: 0000-0003-3910-9727
authors.initials: KD
authors.lastname: Bergmeister
authors.affiliation: Core Facility Imaging, Core Facilities, Medical University Vienna, Austria.
authors.forename: Marion
authors.initials: M
authors.lastname: Gröger
authors.affiliation: CD Laboratory for the Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Spitalgasse 23, A-1090, Austria.
authors.forename: Martin
authors.initials: M
authors.lastname: Aman
authors.affiliation: CD Laboratory for the Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Spitalgasse 23, A-1090, Austria.
authors.forename: Anna
authors.initials: A
authors.lastname: Willensdorfer
authors.affiliation: CD Laboratory for the Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Spitalgasse 23, A-1090, Austria.
authors.forename: Krisztina
authors.initials: K
authors.lastname: Manzano-Szalai
authors.affiliation: CD Laboratory for the Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Spitalgasse 23, A-1090, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Salminger
authors.affiliation: CD Laboratory for the Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Spitalgasse 23, A-1090, Austria.
authors.forename: Oskar C
authors.initials: OC
authors.lastname: Aszmann
mesh_terms: D000818:Animals; D003936:Diagnosis, Computer-Assisted; D003952:Diagnostic Imaging; D007150:Immunohistochemistry; D008297:Male; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000145:classification / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012044:Regression Analysis
chemical_list: D018995:Myosin Heavy Chains
keywords: ImageJ; automated muscle fiber analysis; fiber population; muscle fiber type; myosin heavy chain; rat