MEASUREMENT OF COLOCALIZATION OF OBJECTS IN DUAL-COLOR CONFOCAL IMAGES
MEASUREMENT OF COLOCALIZATION OF OBJECTS IN DUAL-COLOR CONFOCAL IMAGES
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DOI：
10.1111/j.1365-2818.1993.tb03313.x
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发表时间：
1993-03-01
期刊：
影响因子：
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通讯作者：
ATEN, JA
中科院分区：
文献类型：
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作者：
MANDERS, EMM;VERBEEK, FJ;ATEN, JA
提出了一种测量共聚焦双色图像中物体共定位程度的方法。该图像分析产生了两个系数，表示双通道图像中每个分量中共定位对象的比例。生成具有高斯强度分布的测试对象，在双通道图像的两个组件的明确位置，允许对程序的可靠性进行准确的调查。为此，在使用背景、串扰和泊松噪声对图像进行降噪之前，先确定共定位系数。这些合成的图像劣化源代表了在实际共聚焦成像中必须处理的劣化源，即暗电流、荧光探针的非特异性结合和交叉反应性、光串扰和光子噪声。通过滤波和串扰校正对退化图像进行恢复。恢复图像的共定位系数与未退化图像的共定位系数差异不显著。最后，我们在真实生物标本的图像上对该程序进行了测试。这些试验的结果与文献中的数据相符。我们得出结论，共定位系数可以提供生物物体或过程之间位置关系的相关定量信息。
A method to measure the degree of co-localization of objects in confocal dual-colour images has been developed. This image analysis produced two coefficients that represent the fraction of co-localizing objects in each component of a dual-channel image. The generation of test objects with a Gaussian intensity distribution, at well-defined positions in both components of dual-channel images, allowed an accurate investigation of the reliability of the procedure. To do that, the co-localization coefficients were determined before degrading the image with background, cross-talk and Poisson noise. These synthesized sources of image deterioration represent sources of deterioration that must be dealt with in practical confocal imaging, namely dark current, non-specific binding and cross-reactivity of fluorescent probes, optical cross-talk and photon noise. The degraded images were restored by filtering and cross-talk correction. The co-localization coefficients of the restored images were not significantly different from those of the original undegraded images. Finally, we tested the procedure on images of real biological specimens. The results of these tests correspond with data found in the literature. We conclude that the co-localization coefficients can provide relevant quantitative information about the positional relation between biological objects or processes.