Introduction: Skeletal muscle is composed of muscle fibers that are huge and multinuclear cells. Several muscle fibers are surrounded by a perimysium, membranous structure. In this study, we developed a method observing perimysium using a combination of SEM, immunofluorescent staining and picro-Sirius red staining, and checked the validity of the method.
Materials and methods: The soleus muscles of ICR mice were used for observation. Serial paraffin sections were made and one section was observed under SEM. The paraffin section, serial to the SEM specimen, was stained with anti-collagen type IV antibody (a marker for basement membrane). After the observation, the same section was stained using the picro-Sirius red staining method.
Results: Using a stereo pair of SEM images, we confirmed that the perimysium shows a plate-shaped layer. The plate-shaped layer was stained red with the picro-Sirius red staining method, but was negative for anti-collagen type IV antibody.
Discussion: In this study, the three-dimensional image of the perimysium of SEM showed a plate-shaped layer that is consistent with those of prior studies. Moreover, the perimysium stained red with the picro-Sirius red staining method is also consistent with those of prior studies. In this study, the perimysium was negative for anti-collagen type IV antibody (a marker for basement membrane). This means that the structure recognized as perimysium is not an ablated basement membrane, artifact. Based on these results, our method was confirmed as an appropriate technique for observing the perimysium. Using our methods, the perimysium was easily visualized in detail.
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