Eaton Samantha L, Cumyn Elizabeth, King Declan, Kline Rachel A, Carpanini Sarah M, Del-Pozo Jorge, Barron Rona, Wishart Thomas M
Roslin Institute, University of Edinburgh, Edinburgh, UK.
Royal (Dick) School of Veterinary Studies, University of Edinburgh, Edinburgh, UK.
J Anat. 2016 Jan;228(1):203-13. doi: 10.1111/joa.12398. Epub 2015 Oct 29.
Quantification of immunohistochemically (IHC) labelled tissue sections typically yields semi-quantitative results. Visualising infrared (IR) 'tags', with an appropriate scanner, provides an alternative system where the linear nature of the IR fluorophore emittance enables realistic quantitative fluorescence IHC (QFIHC). Importantly, this new technology enables entire tissue sections to be scanned, allowing accurate area and protein abundance measurements to be calculated from rapidly acquired images. Here, some of the potential benefits of using IR-based tissue imaging are examined, and the following are demonstrated. Firstly, image capture and analysis using IR-based scanning technology yields comparable area-based quantification to those obtained from a modern high-resolution digital slide scanner. Secondly, IR-based dual target visualisation and expression-based quantification is rapid and simple. Thirdly, IR-based relative protein abundance QIHC measurements are an accurate reflection of tissue sample protein abundance, as demonstrated by comparison with quantitative fluorescent Western blotting data. In summary, it is proposed that IR-based QFIHC provides an alternative method of rapid whole-tissue section low-resolution imaging for the production of reliable and accurate quantitative data.
免疫组织化学（IHC）标记的组织切片定量通常会产生半定量结果。使用合适的扫描仪对红外（IR）“标签”进行可视化，提供了一种替代系统，其中IR荧光团发射的线性特性使得能够进行实际的定量荧光免疫组织化学（QFIHC）。重要的是，这项新技术能够对整个组织切片进行扫描，从而可以根据快速获取的图像计算出准确的面积和蛋白质丰度测量值。在此，我们研究了使用基于IR的组织成像的一些潜在益处，并展示了以下内容。首先，使用基于IR的扫描技术进行图像捕获和分析，所获得的基于面积的定量结果与现代高分辨率数字载玻片扫描仪获得的结果相当。其次，基于IR的双靶点可视化和基于表达的定量快速且简单。第三，与定量荧光蛋白质印迹数据相比表明，基于IR的相对蛋白质丰度QIHC测量准确反映了组织样本中的蛋白质丰度。总之，有人提出基于IR的QFIHC为生成可靠且准确的定量数据提供了一种快速全组织切片低分辨率成像的替代方法。
