A new web-based method for automated analysis of muscle histology

BMC Musculoskeletal Disorders

Technical Advance

Hanns Lochmüller¹  Stefan Pfeifer²  Erich Wintermantel²  Markus Eblenkamp³  Maggie C Walter⁴  Cordula Pertl⁴  Anja Pertl⁴  Sabine Krause⁴  Christian Thirion⁵ 

[1] Institute of Genetic Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom;Institute of Medical and Polymer Engineering, Technische Universität München, Munich, Germany;Institute of Medical and Polymer Engineering, Technische Universität München, Munich, Germany;S.CO LifeScience GmbH, Munich, Germany;Laboratory of Molecular Myology, Friedrich-Baur-Institute, Department of Neurology, Ludwig-Maximilians-Universität München, Marchioninistraße 17, 81377, Munich, Germany;SIRION BIOTECH GmbH, Planegg/Martinsried, Germany;

关键词: Duchenne muscular dystrophy;    md;    Histological muscle fibre analysis;    Standardised and automated quantitative analysis;    Minimal Feret’s diameter;   

DOI  :  10.1186/1471-2474-14-26

 received in 2012-06-18, accepted in 2013-01-13,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundDuchenne Muscular Dystrophy is an inherited degenerative neuromuscular disease characterised by rapidly progressive muscle weakness. Currently, curative treatment is not available. Approaches for new treatments that improve muscle strength and quality of life depend on preclinical testing in animal models. The mdx mouse model is the most frequently used animal model for preclinical studies in muscular dystrophy research. Standardised pathology-relevant parameters of dystrophic muscle in mdx mice for histological analysis have been developed in international, collaborative efforts, but automation has not been accessible to most research groups. A standardised and mainly automated quantitative assessment of histopathological parameters in the mdx mouse model is desirable to allow an objective comparison between laboratories.MethodsImmunological and histochemical reactions were used to obtain a double staining for fast and slow myosin. Additionally, fluorescence staining of the myofibre membranes allows defining the minimal Feret’s diameter. The staining of myonuclei with the fluorescence dye bisbenzimide H was utilised to identify nuclei located internally within myofibres. Relevant structures were extracted from the image as single objects and assigned to different object classes using web-based image analysis (MyoScan). Quantitative and morphometric data were analysed, e.g. the number of nuclei per fibre and minimal Feret’s diameter in 6 month old wild-type C57BL/10 mice and mdx mice.ResultsIn the current version of the module “MyoScan”, essential parameters for histologic analysis of muscle sections were implemented including the minimal Feret’s diameter of the myofibres and the automated calculation of the percentage of internally nucleated myofibres. Morphometric data obtained in the present study were in good agreement with previously reported data in the literature and with data obtained from manual analysis.ConclusionsA standardised and mainly automated quantitative assessment of histopathological parameters in the mdx mouse model is now available. Automated analysis of histological parameters is more rapid and less time-consuming. Moreover, results are unbiased and more reliable. Efficacy of therapeutic interventions, e.g. within the scope of a drug screening or therapeutic exon skipping, can be monitored. The automatic analysis system MyoScan used in this study is not limited exclusively to dystrophin-deficient mice but also represents a useful tool for applications in the research of other dystrophic pathologies, various other skeletal muscle diseases and degenerative neuromuscular disorders.

【 授权许可】

CC BY   
© Pertl et al.; licensee BioMed Central Ltd. 2013