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Biomed Opt Express. 2017 May 15;8(6):2965-2975. doi: 10.1364/BOE.8.002965. eCollection 2017 Jun 1.

Photothermal imaging of skeletal muscle mitochondria.

Author information

1
Advanced Ultrafast Laser Research Center, The University of Electro-Communications, 1-5-1 Chofugaoka, Chofu, Tokyo 182-8585, Japan.
2
Brain Science Inspired Life Support Research Center, The University of Electro-Communications, 1-5-1 Chofugaoka, Chofu, Tokyo 182-8585, Japan.
3
Advanced Ultrafast Laser Research Center, Department of Electrophysics, National Chiao-Tung University, 1001 Ta Hsinchu Rd., Hsinchu 300, Taiwan.

Abstract

The morphology and topology of mitochondria provide useful information about the physiological function of skeletal muscle. Previous studies of skeletal muscle mitochondria are based on observation with transmission, scanning electron microscopy or fluorescence microscopy. In contrast, photothermal (PT) microscopy has advantages over the above commonly used microscopic techniques because of no requirement for complex sample preparation by fixation or fluorescent-dye staining. Here, we employed the PT technique using a simple diode laser to visualize skeletal muscle mitochondria in unstained and stained tissues. The fine mitochondrial network structures in muscle fibers could be imaged with the PT imaging system, even in unstained tissues. PT imaging of tissues stained with toluidine blue revealed the structures of subsarcolemmal (SS) and intermyofibrillar (IMF) mitochondria and the swelling behavior of mitochondria in damaged muscle fibers with sufficient image quality. PT image analyses based on fast Fourier transform (FFT) and Grey-level co-occurrence matrix (GLCM) were performed to derive the characteristic size of mitochondria and to discriminate the image patterns of normal and damaged fibers.

KEYWORDS:

(170.0110) Imaging systems; (170.0170) Medical optics and biotechnology; (350.5340) Photothermal effects

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