Muscle Histology Characterization Using H&E Staining and Muscle Fiber Type Classification Using Immunofluorescence Staining

• Published in: Bio-protocol Vol. 7; no. 10
• Main Authors: Wang, Chao, Yue, Feng, Kuang, Shihuan
• Format: Journal Article
• Language: English
• Published: United States Bio-Protocol 20.05.2017
• Subjects: Biology; Methods; Myofiber type; Histology; H&E staining; Immunostaining; Skeletal muscle
• ISSN: 2331-8325; 2331-8325
• DOI: 10.21769/bioprotoc.2279

Muscle function is determined by its structure and fiber type composition. Here we describe a protocol to examine muscle histology and myofiber types using hematoxylin and eosin (H&E) and immunofluorescence staining, respectively. H&E stain nucleus in blue and cytoplasm in red, therefore allowing for morphological analyses, such as myofiber diameter, the presence of degenerated and regenerated myofibers, and adipocytes and fibrotic cells. Muscle fibers in adult skeletal muscles of rodents are classified into 4 subtypes based on the expression of myosin heavy chain proteins: Myh7 (type I fiber), Myh2 (type IIA fiber), Myh1 (type IIX fiber), Myh4 (type IIB fiber). A panel of monoclonal antibodies can be used to specifically label these muscle fiber subtypes. These protocols are commonly used in the study of muscle development, growth and regeneration (for example: Wang , 2015; Nie , 2016; Yue , 2016; Wang , 2017).