Statistical and Economical Efficiency in Assessment of Liver Regeneration Using Defined Sample Size and Selection in Combination With a Fully Automated Image Analysis System
Statistical and Economical Efficiency in Assessment of Liver Regeneration Using Defined Sample Size and Selection in Combination With a Fully Automated Image Analysis System
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DOI：
10.1369/jhc.2009.953869
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发表时间：
2009-11-01
影响因子：
3.2
通讯作者：
Dahmen, Uta
中科院分区：
文献类型：
--
作者：
Deng, Meihong;Kleinert, Robert;Dahmen, Uta
肝再生的定量通常基于确定5-溴-2-脱氧尿苷标记指数（BrdU-LI）。定量结果受分析前、分析后和分析后变量（如感兴趣区域（ROI））的影响。我们的目的是介绍我们新开发和验证的自动计算机图像分析系统（AnalySIS-Macro），并标准化ROI的选择和样本量。对BrdU标记和化学染色肝脏切片的图像进行常规分析，并使用新开发的AnalysIS-Macro进行分析，并用于系统验证。自动定量与人工计数结果相关性良好（r=0.9976）。我们的AnalysIS-Macro验证显示其高灵敏度（>90%）和特异性。同一肝脏内BrdU-LI范围为11%-57%（32.96 +/- 11.94%），反映了肝细胞增殖的高度可变空间分布。每个肝叶至少需要2000个肝细胞（10个图像，200倍放大）作为样本量，以获得代表性BrdU-LI。此外，中央周围区的数目应等于门静脉周围区的数目。我们的AnalysIS-Macro与ROI选择和大小规则的组合代表了用于测定BrdU-LI和增殖肝细胞空间分布的准确、灵敏、特异和有效的诊断工具。（J Histochem Cytochem 57：1075-1085，2009）
Quantification of liver regeneration is frequently based on determining the 5-bromo-2-deoxyuridine labeling index (BrdU-LI). The quantitative result is influenced by preanalytical, analytical, and postanalytical variables such as the region of interest (ROI). We aimed to present our newly developed and validated automatic computer-based image analysis system (AnalySIS-Macro), and to standardize the selection and sample size of ROIs. Images from BrdU-labeled and immunohistochemically stained liver sections were analyzed conventionally and with the newly developed AnalySIS-Macro and used for validation of the system. Automatic quantification correlated well with the manual counting result (r=0.9976). Validation of our AnalySIS-Macro revealed its high sensitivity (>90%) and specificity. The BrdU-LI ranged from 11% to 57% within the same liver (32.96 +/- 11.94%), reflecting the highly variable spatial distribution of hepatocyte proliferation. At least 2000 hepatocytes(10 images at 200 x magnification) per lobe were required as sample size for achieving a representative BrdU-LI. Furthermore, the number of pericentral areas should be equal to that of periportal areas. The combination of our AnalySIS-Macro with rules for the selection and size of ROIs represents an accurate, sensitive, specific, and efficient diagnostic tool for the determination of the BrdU-LI and the spatial distribution of proliferating hepatocytes. (J Histochem Cytochem 57:1075-1085, 2009)