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

Episode von Medizin - Open Access LMU - Teil 21/22 von Ludwig-Maximilians-Universität München · 1. Januar 2013

Background: Duchenne 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.

Methods: Immunological 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.

Results: In 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.

Conclusions: A 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.

Weitere Episoden von „Medizin - Open Access LMU - Teil 21/22“

• A genotype-specific, randomized controlled behavioral intervention to improve the neuroemotional outcome of cardiac surgery: study protocol for a randomized controlled trial 1. Januar 2013
• Acceptance of routine or case-based inquiry for intimate partner violence: a mixed method study 1. Januar 2013
• Activation of axonal Kv7 channels in human peripheral nerve by flupirtine but not placebo - therapeutic potential for peripheral neuropathies: results of a randomised controlled trial 1. Januar 2013
• Aequilibrium prudentis: on the necessity for ethics and policy studies in the scientific and technological education of medical professionals 1. Januar 2013
• An AUC-based permutation variable importance measure for random forests 1. Januar 2013
• Autosomal recessive spastic ataxia of Charlevoix Saguenay (ARSACS): expanding the genetic, clinical and imaging spectrum 1. Januar 2013
• `Burden to others' as a public concern in advanced cancer: a comparative survey in seven European countries 1. Januar 2013
• Characterization of in vivo chemoresistant human hepatocellular carcinoma cells with transendothelial differentiation capacities 1. Januar 2013
• Conscious Presence and Self Control as a measure of situational awareness in soldiers - A validation study 1. Januar 2013
• Contrast enhanced transesophageal echocardiography in patients with atrial fibrillation referred to electrical cardioversion improves atrial thrombus detection and may reduce associated thromboembolic events 1. Januar 2013
• Current experience with applying the GRADE approach to public health interventions: an empirical study 1. Januar 2013
• Current management of patients hospitalized with community-acquired pneumonia across Europe: outcomes from REACH 1. Januar 2013
• Diversity of the var gene family of Indonesian Plasmodium falciparum isolates 1. Januar 2013
• Driving forces for changes in geographical distribution of Ixodes ricinus ticks in Europe 1. Januar 2013
• A comparative analysis of the properties of regulated promoter systems commonly used for recombinant gene expression in Escherichia coli 1. Januar 2013

Alle Episoden von Medizin - Open Access LMU - Teil 21/22 · Mehr Bildung-Podcasts