A double fluorescence staining protocol to determine the cross-sectional area of myofibers using image analysis
A double fluorescence staining protocol to determine the cross-sectional area of myofibers using image analysis
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DOI：
10.3109/10520299609117143
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发表时间：
1996-03-01
影响因子：
1.6
通讯作者：
Cassens, RG
中科院分区：
文献类型：
--
作者：
Mozdziak, PE;Fassel, TA;Cassens, RG
开发了一种双荧光染色方案，以便于基于计算机的图像分析，来自实验处理的肌纤维用抗肌球蛋白抗体MF-20标记（辐照的）和对照生长的火鸡骨骼肌，并用荧光素-5-异硫氰酸酯（FITC）检测。用伴刀豆球蛋白A（ConA）-德克萨斯红染色细胞外材料，从MF-20-FITC图像中减去每个感兴趣区域的ConA-Texas红色图像后，通过计算覆盖每个肌纤维的像素数（0.83 μ m（2））来确定肌纤维的横截面积。与预期的一样，照射肌肉中的肌纤维比未照射肌肉中的肌纤维小（P < 0.05）。这种双荧光染色结合图像分析的方案是准确的，并且比用于确定肌纤维横截面积的经典方法劳动强度更小。
A double fluorescence staining protocol was developed to facilitate computer based image analysis, Myofibers from experimentally treated (irradiated) and control growing turkey skeletal muscle were labeled with the anti-myosin antibody MF-20 and detected using fluorescein-5-isothiocyanate (FITC), Extracellular material was stained with concanavalin A (ConA)-Texas red, The cross-sectional area of the myofibers was determined by calculating the number of pixels (0.83 mu m(2)) overlying each myofiber after subtracting the ConA-Texas red image from the MF-20-FITC image for each region of interest. As expected, myofibers in the irradiated muscle were smaller (P < 0.05) than those in the non-irradiated muscle, This double fluorescence staining protocol combined with image analysis is accurate and less labor-intensive than classical procedures for determining the cross-sectional area of myofibers.