Lipid imaging of human skeletal muscle using TOF-SIMS with bismuth cluster ion as a primary ion source.
Journal article, 2008
Intramyocellular lipids are of importance in lipid-related diseases. The techniques in this field are limited because of a lack of adequate tools for localization of various lipids. The most usual methods for the localization of lipid distribution in the skeletal muscle are histochemistry and fluorescence probes. Different chromatography methods and mass spectrometry techniques have also been used for lipid identification. Our aim was to localize the spatial distribution of lipids in their native forms by using static time-of-flight secondary-ion mass spectrometry (TOF-SIMS). Human percutaneous skeletal muscle biopsies were obtained from the middle part of the lateral vastus muscle in the right leg of healthy adolescents with a body mass index >30. Samples were prepared by high-pressure freezing, freeze-fracturing and freeze-drying, and analysed by imaging TOF-SIMS equipped with a Bi3+ cluster ion gun. In the positive spectra, we identified phosphocholine, cholesterol, diacylglycerol, phospholipids and triacylglycerol. Phosphocholine was localized to the edge of the fibre, representing the sarcoplasma or endomysium. Weak cholesterol signals were observed in the intracellular areas. High diacylglycerol and low triacylglycerol signal intensities were seen in intracellular spaces of the transversal area of the muscle fibre. In the negative spectra, we identified fatty acids. We observed co-localization of fatty acids and diacylglycerol, which may indicate lipid-storing parts of the skeletal muscle. Thus, TOF-SIMS imaging can be used to depict the heterogeneous localization of lipids in human skeletal muscle.
analysis
Freeze Drying
methods
Mass
Freeze Fracturing
Secondary Ion
Cholesterol
Adolescent
analysis
analysis
Diglycerides
Microscopy
Phosphorylcholine
analysis
ultrastructure
Fast-Twitch
Slow-Twitch
ultrastructure
chemistry
Fatty Acids
analysis
Scanning
Spectrometry
Humans
Lipids
Muscle Fibers
Quadriceps Muscle
Bismuth
chemistry
Muscle Fibers
ultrastructure
Electron
Triglycerides
chemistry
analysis