Zinchuk Vadim, Zinchuk Olga, Okada Teruhiko
Department of Anatomy and Cell Biology, Kochi University Faculty of Medicine, Okoh-cho, Nankoku, Kochi 783-8505, Japan.
Acta Histochem Cytochem. 2007 Aug 30;40(4):101-11. doi: 10.1267/ahc.07002.
Quantitative colocalization analysis is an advanced digital imaging tool to examine antigens of interest in immunofluorescence images obtained using confocal microscopes. It employs specialized algorithms to estimate the degree of overlap of fluorescence signals and thus enables acquiring important new information not otherwise obtainable using qualitative approaches alone. As raw confocal images have high levels of background, they should be prepared to become suitable for reliable calculation of colocalization coefficients by correcting it. We provide concise theoretical basis of quantitative colocalization analysis, discuss its limitations, and describe proper use of the technique. The use of quantitative colocalization analysis is demonstrated by studying bile salt export pump and multidrug resistance associated protein 2 in the liver and major basic protein and platelet activating factor receptor antigens in conjunctiva. The review is focused on the applicability and correct interpretation of the results of colocalization coefficients calculations.
定量共定位分析是一种先进的数字成像工具，用于检查在使用共聚焦显微镜获得的免疫荧光图像中感兴趣的抗原。它采用专门的算法来估计荧光信号的重叠程度，从而能够获取仅使用定性方法无法获得的重要新信息。由于原始共聚焦图像具有高水平的背景，应通过校正使其适合可靠地计算共定位系数。我们提供了定量共定位分析的简明理论基础，讨论了其局限性，并描述了该技术的正确使用方法。通过研究肝脏中的胆盐输出泵和多药耐药相关蛋白2以及结膜中的主要碱性蛋白和血小板活化因子受体抗原，展示了定量共定位分析的应用。本综述重点关注共定位系数计算结果的适用性和正确解释。
