AUTOMATED MUSCLE FIBER TYPE POPULATION ANALYSIS WITH IMAGEJ OF WHOLE RAT MUSCLES USING RAPID MYOSIN HEAVY CHAIN IMMUNOHISTOCHEMISTRY
AUTOMATED MUSCLE FIBER TYPE POPULATION ANALYSIS WITH IMAGEJ OF WHOLE RAT MUSCLES USING RAPID MYOSIN HEAVY CHAIN IMMUNOHISTOCHEMISTRY
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DOI：
10.1002/mus.25033
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发表时间：
2016-08-01
期刊：
影响因子：
3.4
通讯作者：
Aszmann, Oskar C.
中科院分区：
文献类型：
--
作者：
Bergmeister, Konstantin D.;Groeger, Marion;Aszmann, Oskar C.
简介：骨骼肌由不同类型的纤维组成，适应运动，衰老，疾病或创伤。在这里，我们提出了一个快速染色，自动采集和量化的纤维群体与ImageJ协议。方法：从Sprague-Dawley大鼠身上获取肱二头肌和蚯蚓肌。使用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：该方案为自动图像采集提供了快速可靠的染色。人工和自动数据之间的分析表明，皮尔森相关系数二头肌为0.645-0.841和0.564-0.673蚓状肌。相对纤维种群的准确度为+/-4%。结论：该协议提供了一个可靠的工具，定量肌纤维种群。使用免费的软件，它减少了分析整个肌肉切片所需的时间。
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.