Open-CSAM, a new tool for semi-automated analysis of myofiber cross-sectional area in regenerating adult skeletal muscle
Open-CSAM, a new tool for semi-automated analysis of myofiber cross-sectional area in regenerating adult skeletal muscle
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DOI：
10.1186/s13395-018-0186-6
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发表时间：
2019-01-08
期刊：
影响因子：
4.9
通讯作者：
Juban, Gaetan
中科院分区：
文献类型：
--
作者：
Desgeorges, Thibaut;Liot, Sophie;Juban, Gaetan
成人骨骼肌在急性损伤后能够完全再生。评估肌肉再生效果的主要参数是肌纤维的横截面积(CSA)，因为肌纤维的大小与肌力相关。CSA分析可能很耗时，并且在手动执行时可能会引发结果的变异性。这就是为什么开发程序来完全自动化CSA的分析，如SMASH、MyoVision或MuscleJ软件。虽然这些软件可以有效地测量正常肌肉或肥大/萎缩肌肉的CSA，但它们无法有效地测量再生肌肉的CSA。我们开发了一种名为Open-CSAM的ImageJ宏，用于在不同的实验条件下对骨骼肌上的CSA进行高通量半自动分析。宏允许实验者调整分析并纠正自动化所犯的错误，这在全自动化程序中是不可能的。我们发现，与SMASH、MyoVision和MuscleJ软件相比，Open-CSAM在测量再生肌肉和营养不良肌肉中的CSA时更准确，而且实验者之间的变异性可以忽略不计。我们还表明，为了获得具有代表性的CSA测量，有必要分析整个肌肉切片，而不是随机选择图片，这一过程使用Open-CSAM可以轻松而准确地执行。总之，我们在这里展示了一种简单且由实验者控制的工具，可以在任何实验条件下测量肌肉中的CSA，包括再生肌肉。
Adult skeletal muscle is capable of complete regeneration after an acute injury. The main parameter studied to assess muscle regeneration efficacy is the cross-sectional area (CSA) of the myofibers as myofiber size correlates with muscle force. CSA analysis can be time-consuming and may trigger variability in the results when performed manually. This is why programs were developed to completely automate the analysis of the CSA, such as SMASH, MyoVision, or MuscleJ softwares. Although these softwares are efficient to measure CSA on normal or hypertrophic/atrophic muscle, they fail to efficiently measure CSA on regenerating muscles. We developed Open-CSAM, an ImageJ macro, to perform a high throughput semi-automated analysis of CSA on skeletal muscle from various experimental conditions. The macro allows the experimenter to adjust the analysis and correct the mistakes done by the automation, which is not possible with fully automated programs. We showed that Open-CSAM was more accurate to measure CSA in regenerating and dystrophic muscles as compared with SMASH, MyoVision, and MuscleJ softwares and that the inter-experimenter variability was negligible. We also showed that, to obtain a representative CSA measurement, it was necessary to analyze the whole muscle section and not randomly selected pictures, a process that was easily and accurately be performed using Open-CSAM. To conclude, we show here an easy and experimenter-controlled tool to measure CSA in muscles from any experimental condition, including regenerating muscle.