使用快速肌球蛋白重链免疫组织化学方法，通过ImageJ对全大鼠肌肉进行自动肌纤维类型群体分析。
Automated muscle fiber type population analysis with ImageJ of whole rat muscles using rapid myosin heavy chain immunohistochemistry.
作者信息
Bergmeister Konstantin D, Gröger Marion, Aman Martin, Willensdorfer Anna, Manzano-Szalai Krisztina, Salminger Stefan, Aszmann Oskar C
机构信息
CD Laboratory for the Restoration of Extremity Function, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University of Vienna, Spitalgasse 23, A-1090, Austria.
Core Facility Imaging, Core Facilities, Medical University Vienna, Austria.
出版信息
Muscle Nerve. 2016 Aug;54(2):292-9. doi: 10.1002/mus.25033. Epub 2016 Jun 13.
INTRODUCTION
Skeletal muscle consists of different fiber types which adapt to exercise, aging, disease, or trauma. Here we present a protocol for fast staining, automatic acquisition, and quantification of fiber populations with ImageJ.
METHODS
Biceps and lumbrical muscles were harvested from Sprague-Dawley rats. Quadruple immunohistochemical staining was performed on single sections using antibodies against myosin heavy chains and secondary fluorescent antibodies. Slides were scanned automatically with a slide scanner. Manual and automatic analyses were performed and compared statistically.
RESULTS
The protocol provided rapid and reliable staining for automated image acquisition. Analyses between manual and automatic data indicated Pearson correlation coefficients for biceps of 0.645-0.841 and 0.564-0.673 for lumbrical muscles. Relative fiber populations were accurate to a degree of ± 4%.
CONCLUSIONS
This protocol provides a reliable tool for quantification of muscle fiber populations. Using freely available software, it decreases the required time to analyze whole muscle sections. Muscle Nerve 54: 292-299, 2016.
引言
骨骼肌由不同类型的纤维组成，这些纤维会适应运动、衰老、疾病或创伤。在此，我们介绍一种使用ImageJ对纤维群体进行快速染色、自动采集和定量分析的方法。
方法
从Sprague-Dawley大鼠身上获取肱二头肌和蚓状肌。使用抗肌球蛋白重链抗体和二级荧光抗体对单个切片进行四重免疫组织化学染色。用载玻片扫描仪自动扫描载玻片。进行手动和自动分析并进行统计学比较。
结果
该方法为自动图像采集提供了快速且可靠的染色。手动和自动数据之间的分析表明，肱二头肌的皮尔逊相关系数为0.645 - 0.841，蚓状肌为0.564 - 0.673。相对纤维群体的误差在±4%以内。
结论
该方法为肌肉纤维群体的定量分析提供了可靠工具。使用免费软件，它减少了分析整个肌肉切片所需的时间。《肌肉与神经》54: 292 - 299, 2016年。
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