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Although several methodological approaches have been used to study these adaptive responses at the muscle fiber level, one of the most commonly used techniques is immunohistochemistry. Skeletal muscle is a plastic tissue that rapidly adjusts to external demands and environmental stimuli such as exercise, inactivity, and disease ( 9, 13). Thus, the Muscle2View is a viable tool for researchers aiming to quantify immunohistochemical variables from skeletal muscle biopsies. In addition to fiber typing, myonuclei counting, and the quantification of fiber type-specific morphological measurements, the Muscle2View pipeline can identify the complex capillary-to-fiber network from a batch of images within minutes. NEW & NOTEWORTHY Here, we developed a freely available CellProfiler-based pipeline termed Muscle2View, which provides unbiased, high-content analysis of muscle cross-sectional immunohistochemistry images. #Cellprofiler load time series multi page tiff softwareThis robust analysis is done in one single run within a user-friendly and flexible environment based on the free and widely used image software CellProfiler. In addition to the classical morphological measurements, the Muscle2View can identify the complex capillary-to-fiber network and myonuclear density in a fiber type-specific manner. Collectively, we demonstrate that the Muscle2View pipeline can provide unbiased and high-content analysis of muscle cross-sectional immunohistochemistry images. #Cellprofiler load time series multi page tiff manualFor several variables, however, there were differences (5–15%) between values computed by manual counting and Muscle2View, suggesting that the methods should not necessarily be used interchangeably. When comparing the Muscle2View pipeline to manual or semiautomatic analysis, overall the results revealed strong correlations. ![]() The novel identification of the capillary-to-fiber interface allowed for the calculation of microvascular factors such as capillary contacts (CC), individual capillary-to-fiber ratio (C/Fi), and capillary-to-fiber perimeter exchange (CFPE) index. Provided that the images are of sufficient quality and the settings are configured for the specific study, the pipeline allows for automatic and unsupervised analysis of fiber borders, myonuclei, capillaries, and morphometric parameters in a fiber type-specific manner from large batches of images in <10 min/tissue sample. Here, we developed Muscle2View, a free CellProfiler-based pipeline that integrates all key fiber-morphological variables, including the novel quantification of the capillary-to-fiber interface, in one single tool. Because manual immunohistochemical analysis of features such as skeletal muscle fiber typing, capillaries, myonuclei, and fiber size-related parameters is time consuming and prone to user subjectivity, automatic computational methods could allow for faster and more objective evaluation. ![]()
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