Validation of computer-assisted, pixel-based analysis of multiple-colour immunofluorescence histology
Validation of computer-assisted, pixel-based analysis of multiple-colour immunofluorescence histology
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DOI：
10.1016/j.jim.2005.05.005
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发表时间：
2005-07-01
影响因子：
2.2
通讯作者：
Bailey, M
中科院分区：
文献类型：
--
作者：
Inman, CF;Rees, LEN;Bailey, M
免疫组织学的发展允许在组织切片上常规同时使用三种单克隆抗体，用不同的荧光染料标记。这种染色可以识别七种不同的细胞群，限制因素是快速，可靠和可重复性分析。未来对计算机辅助数字化图像分析的依赖取决于对人工计数的验证，这通常被视为“黄金标准”。在这项研究中，从正常的猪皮肤、发炎的皮肤和扁桃体切片中数字化图像，同时用三种单克隆抗体染色。通过四次人工计数和基于像素的面积测量来定量染色组合。在单个图像上，自动和手动测量之间的相关性很差。尽管如此，在组织的平均值和方差方面，人工和自动测量之间的一致性很好，两种技术都确定了炎症组织与正常组织的相同变化。此外，基于像素的计数允许组织切片中细胞类型共定位的统计分析。我们得出的结论是，与人工计数相比，自动计数对组织的评估是可以接受的，它更快，并且为观察者的变化提供了更少的机会。我们还证明，该技术适用于使用三种以上的荧光染料，使人工计数基本上不可能。(c) 2005 Elsevier B.V.版权所有
Developments in immunohistology allow the routine simultaneous use on tissue sections of three monoclonal antibodies, tagged with different fluorochromes. Such staining can identify seven different cell populations and the limiting factor is rapid, reliable and reproducible analysis. Future reliance on computer-assisted analysis of digitised images depends on validation against manual counting, often viewed as the 'gold standard'.In this study images were digitised from sections of normal porcine skin, inflamed skin and tonsil, simultaneously stained with three monoclonal antibodies. Combinations of staining were quantified by four manual counts and by pixel-based area measurement. On individual images, the correlation between automated and manual measurements was poor. Despite this, the concordance between manual and automated measurements in the means and variances of tissues was good, and both techniques identified the same changes in inflamed versus normal tissues. In addition, pixel-based counting permitted statistical analysis of co-localisation of cell types in tissue sections.We conclude that automated counting is acceptable for the assessment of tissues, is faster and provides less opportunity for observer variation than manual counting. We also demonstrate that the technique is applicable where more than three fluorochromes are used such that manual counting becomes essentially impossible. (c) 2005 Elsevier B.V. All rights reserved.