3D ex vivo imaging and characterization of neuromuscular junctions

Your Needs:
  • Study of neuromuscular pathologies or atrophy models
  • Preclinical evaluation of compound efficacy

Our Solutions:
  • Light sheet microscopy and clearing to visualize neuromuscular junctions on whole muscle in 3D
  • Automated image processing for quantification

General Procedure

Prior to sample collection by Imactiv-3D:
  • In vivo labeling by infusion with a fluorescent toxin
  • Formalin fixation of extracted sample

Image acquisition:
  • Sample clearing
  • 3D light sheet fluorescence microscopy
  • Multi-position acquisition

Image processing and analysis:
  • Quantitative characterization of all types of leg muscle:
    • Estimation of muscle volume
    • Absolute number and density of neuromuscular junctions
  • 3D visualization with surface and volume rendering:
    • Reconstruction of the whole sample
    • Visualization of the spatial distribution of junctions
    • Advanced display using 3D animations

Application example

  • Aim: comparison of a wide range of muscle types in the rat leg in terms of neuromuscular junction density
  • Technical specifications:
    • Whole sample acquisition using 3 to 10 fields of view
    • Number of junctions ranges from 3 000 to 20 000 depending on the muscle type
    • Junction average size is about 60 x 30 x 10 µm³
  • Analysis of the neuromuscular junction density following this procedure on 30 muscles produced interesting results among different types of muscle