Automated Compensation of Light Attenuation in Confocal Microscopy by Exact Histogram Specification
Automated Compensation of Light Attenuation in Confocal Microscopy by Exact Histogram Specification
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DOI：
10.1002/jemt.20767
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发表时间：
2010-03-01
影响因子：
2.5
通讯作者：
Coltuc, Dinu
中科院分区：
文献类型：
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作者：
Stanciu, Stefan G.;Stanciu, George A.;Coltuc, Dinu
共聚焦激光扫描显微镜(CLSM)使我们能够捕获代表样品体积上的光学切片的图像。从不同层获得的图像具有不同的对比度：从标本的更深层获得的图像相对于从最上层获得的图像具有较低的对比度。造成上述影响的主要原因是光通过的体积中包含的原子和分子对光的吸收和散射。此外，光衰减也可能由观察表面的倾斜引起。对于倾斜较大的表面，反射光的方向将与探测器的方向不同。我们提出了一种基于直方图运算的数字图像处理技术，可以用来补偿光衰减的影响。对CLSM得到的图像序列进行精确的直方图规定化和均衡化处理。在这种情况下，为了实现准确的直方图建模，必须在像素之间引入严格的排序。处理后的图像最终将精确地具有指定的直方图，而不是具有与指定形状相似的形状的直方图，这与经典的直方图规范算法的情况相同。讨论了诱导排序的实验结果和理论方面，并对几种直方图建模技术在处理共焦显微镜获得的图像序列方面进行了比较。Microsc.技术资源。2010年，73：165-175。(C)2009年Wiley-Liss，Inc.
Confocal laser scanning microscopy (CLSM) enables us to capture images representing optical sections on the volume of a specimen. The images acquired from different layers have a different contrast: the images obtained from the deeper layers of the specimen will have a lower contrast with respect to the images obtained from the topmost layers. The main reasons responsible for the effects described above are light absorption and scattering by the atoms and molecules contained in the volume through which the light passes. Also light attenuation can be caused by the inclination of the observed surface. In the case of the surfaces that have a steep inclination, the reflected light will have a different direction than the one of the detector. We propose a technique of digital image processing that can be used to compensate the effects of light attenuation based on histogram operations. We process the image series obtained by CLSM by exact histogram specification and equalization. In this case, a strict ordering among pixels must be induced hi order to achieve the exact histogram modeling. The processed images will end up having exactly the specified histogram mid not a histogram with a shape that just resembles to the specified one, as in the case of classical histogram specification algorithms. Experimental results and theoretical aspects of the induced ordering are discussed, as well as a comparison between several histogram modeling techniques with respect to the processing of image series obtained by confocal microscopy. Microsc. Res. Tech. 73:165-175, 2010. (c) 2009 Wiley-Liss, Inc.