Axial organization of muscular myosin identified by the optical and computational pipeline FAMOUS

C. Lefort; M. Chalvidal; F. Baraige; A. Parente; E. Ferrandon; V. Blanquet; H. Massias; L. Magnol; E. Chouzenoux

doi:10.1117/12.2621004

20 May 2022 Axial organization of muscular myosin identified by the optical and computational pipeline FAMOUS

C. Lefort, M. Chalvidal, F. Baraige, A. Parente, E. Ferrandon, V. Blanquet, H. Massias, L. Magnol, E. Chouzenoux

Proceedings Volume 12136, Unconventional Optical Imaging III; 1213602 (2022) https://doi.org/10.1117/12.2621004
Event: SPIE Photonics Europe, 2022, Strasbourg, France

Abstract

In a medical use, ultrastructure of muscle is currently revealed by images of resected samples achieved thanks to Electron Microscopy (EM), requiring freezing and paraffin sections, with a set of histological, molecular and biochemical analyses. The resection, slicing and labelling steps cause an alteration of the phenotypic and volumetric information compared to their initial integrity. Starting from this statement, we have developed an original pipeline resting on the imbrication of an optical and computational strategy for imaging 3D biomedical structures without resorting to slicing, freezing and labelling steps. The assembly of myosin of a whole muscle is revealed thanks to the second harmonic generation (SHG) recorded with a multiphoton microscope, all along the entire 180 μm of thickness of the Extensor digitorum longus (EDL) of a wild mouse. During the SHG recording, the Point-Spread-Function (PSF) of the multiphoton microscope is recorded all along the imaging depth. This step highlights an axial broadening of the PSF while maintaining a constant planar PSF throughout the whole depth of the recordings. Then, a fitting algorithm estimated a mathematical model of the PSF, highlighting its variability into the whole image depth. Finally, a computational image restauration is led thanks to the fast image deblurring algorithm BD3MG accounting for the depth variant PSF. The non-stationary distortions all along the recorded image is employed thus correcting accurately the image distortion. The axial organization of the myosin is revealed for the first time, highlighting tubular organization of myosin into the myofibrils.

Conference Presentation

Citation Download Citation

C. Lefort, M. Chalvidal, F. Baraige, A. Parente, E. Ferrandon, V. Blanquet, H. Massias, L. Magnol, and E. Chouzenoux "Axial organization of muscular myosin identified by the optical and computational pipeline FAMOUS", Proc. SPIE 12136, Unconventional Optical Imaging III, 1213602 (20 May 2022); https://doi.org/10.1117/12.2621004

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