Automatic section thickness determination using an absolute gradient focus function.

Automatic section thickness determination using an absolute gradient focus function.
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DOI:
10.1111/j.1365-2818.2012.03669.x
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发表时间:
2012-12
影响因子:
2
通讯作者:
Mouton PR
Mouton PR
中科院分区:
工程技术4区
文献类型:
--
作者:
Elozory DT;Kramer KA;Chaudhuri B;Bonam OP;Goldgof DB;Hall LO;Mouton PR

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使用计算机体视学系统对微观结构进行定量分析是生物科学研究许多学科的重要工具。当前非自动化方法中的切片厚度确定需要手动定位组织切片的上表面和下表面。与使用聚焦曲线上的全局最大值来定位最佳聚焦光学平面的传统自动聚焦功能相比,本研究通过聚焦曲线上的两个尖锐“拐点”来识别从未聚焦到聚焦光学平面的过渡。对十四个灰度聚焦函数的分析表明,阈值绝对梯度函数最适合寻找与上下组织表面的边界光学平面紧密对应的可检测弯曲。对该函数的修改产生了四个优于原始函数的新函数。 “修改后的绝对梯度计数”函数在与训练集相似的图像测试集上的平均误差为 0.56 μm,优于所有其他函数;并且,在由从不同病例捕获的图像组成的测试集上的平均误差为 0.39 μm,即对不同受试者大鼠的不同大脑区域采用不同的染色方法。我们描述了一种新颖的算法,它允许基于离焦平面自动确定切片厚度,这是全自动计算机体视学的先决条件。
Quantitative analysis of microstructures using computerized stereology systems is an essential tool in many disciplines of bioscience research. Section thickness determination in current non-automated approaches requires manual location of upper and lower surfaces of tissue sections. In contrast to conventional autofocus functions that locate the optimally focused optical plane using the global maximum on a focus curve, the present study identified by two sharp “knees” on the focus curve as the transition from unfocused to focused optical planes. Analysis of fourteen gray-scale focus functions showed, the thresholded absolute gradient function, was best for finding detectable bends that closely correspond to the bounding optical planes at the upper and lower tissue surfaces. Modifications to this function generated four novel functions that outperformed the original. The “modified absolute gradient count” function outperformed all others with an average error of 0.56 μm on a test set of images similar to the training set; and, an average error of 0.39 μm on a test set comprised of images captured from a different case, i.e., different staining methods on a different brain region from a different subject rat. We describe a novel algorithm that allows for automatic section thickness determination based on just out-of-focus planes, a prerequisite for fully automatic computerized stereology.
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