Muscle Fibre IHC Analysis

Determination of individual muscle fibre parameters and types within tissues can generate valuable quantitative information on changes during various diseases and/or drug treatments. Neuromuscular disease encompasses many diseases and ailments that impair the functioning of the muscles, either directly, being pathologies of the muscle, or indirectly, being pathologies of nerves or neuromuscular junctions. Muscular dystrophies, including Duchenne’s and Becker’s, are a large group of diseases, many of them hereditary or resulting from genetic mutations, where the muscle integrity is disrupted.

Left: Muscle tissue stained with spectrin (white) and Type IIB fibers (green). Right: Detection of Type IIB (purple) and Type I/IIA (blue) individual fibers

Analysis Approach

Immunofluoresence and IHC can be used to highlight different parameters in the tissue including the cytoskeletal protein spectrin and individual muscle fibre types. Digital image analysis can then be used to determine the structure of individual fibres and quantify various parameters. In the example shown, a customised image analysis algorithm is developed to detect the spectrin layer (yellow overlay), which defines the shape of each muscle fibre object. Fibre types are then automatically classified as either Type IIB (green stained, purple overlay) or Type I/IIA (unstained, blue overlay). Finally, each tissue slide was manually reviewed for exclusion of detected artefacts (grey overlay) before the cross sectional area (µm2) of each classified fibre was determined.

Quantitative Readouts

  • Number of individual muscle fibres present in whole tissue section
  • Classification of muscle fibres based on cross sectional area, length, circumference, shape index etc
  • Classification of muscle fibre types based on IF staining (i.e. Type II, Type I/IIA)

Left: Whole muscle tissue stained with spectrin (white) and Type IIB fibers (green). Right: Detection of Type IIB (purple) and Type I/IIA (blue) individual fibers across whole section

Benefits

  • Receive highly detailed data quantifying morphological features of individual muscle fibres across a whole tissue section or within defined regions of interest.
  • Our automated batch analysis service objectively analyses each section using the same algorithm to reduce variability and improve data quality and interpretation