Protocol to segment and characterize the morphometry of individual cross-sectioned striated muscle fibers from adult murine models

Muscle fibre morphometric analysis (MusMA) correlates with muscle function and cardiovascular risk prognosis

“…The semiautomated MusMA tool used in this study can provide an accurate screening of muscle health by providing a complete description of the size and shape of muscle fibres. 24 This analysis was performed using MusMA, which provided an overview of the health of skeletal and cardiac muscle tissue through the separation of muscle fibres into six subpopulations, namely, normal (regular) fibres, regular hypertrophic, irregular hypertrophic, irregular, irregular atrophic and regular atrophic. Based on this segmentation, we found associations between biological and molecular data from different experimental models.…”

“…As main points of attention, we can mention the quality of the histological sections, the staining of the slides with haematoxylin and eosin and the condition of the acquired images. 24 It should be noted that in the literature, the significant variability of labelling methods and sorting of fibre morphometry can interfere with results obtained.…”

“…The MusMA evaluation technique was developed by our research group based on Nuclear Morphometric Analysis (NMA), to obtain information about the size and shape of the cell nucleus in vitro and in tissue stained with haematoxylin and eosin 35,36 . The MusMA protocol was described in detail in a previous publication 24 . Briefly, transversally sectioned muscle fibres from skeletal striated muscle and cardiac striated muscle stained with haematoxylin and eosin were segmented using Image Pro Plus 6.0, followed by the acquisition of size and shape measurements for individual cells.…”

“…35,36 The MusMA protocol was described in detail in a previous publication. 24 Briefly, transversally sectioned muscle fibres from skeletal striated muscle and cardiac striated muscle stained with haematoxylin and eosin were segmented using Image Pro Plus 6.0, followed by the acquisition of size and shape measurements for individual cells. The variable 'area' was obtained to assess the size of each muscle fibre.…”

“…However, several of these techniques are complex, expensive, or limited to measuring fibre area 20–23 . Recently, we developed a new method to evaluate muscle fibre size and shape, the Muscle Fibre Morphometric Analysis (MusMA) 24 . Based on the MusMA method, in this study, we associated morphometry data obtained by using MusMA with parameters of skeletal muscle function and cardiovascular risk.…”