Extended depth of focus in optical microscopy: Assessment of existing methods and a new proposal
Extended depth of focus in optical microscopy: Assessment of existing methods and a new proposal
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DOI：
10.1002/jemt.22104
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发表时间：
2012-11-01
影响因子：
2.5
通讯作者：
Bevilacqua, Alessandro
中科院分区：
文献类型：
--
作者：
Piccinini, Filippo;Tesei, Anna;Bevilacqua, Alessandro
由于焦深的限制，使用标准光学显微镜不可能获得由相关深度维度表征的单个2-D完全对焦的3-D对象的图像。自七十年代以来，已经提出了许多方法来克服这个问题，主要是通过不同的融合处理技术来延长显微镜的焦深。然而，对于特定的应用，很难知道哪种方法产生最好的结果，因为没有经过验证的方法或测试的指标适用于现实世界中通常缺乏参考基本事实的情况。虽然通用质量指数(UQI)在图像处理中被广泛用于评估输出质量，但它需要参考地面事实。已经提出了一些UQI扩展来评估没有基本事实的融合方法的输出，但还没有进行足够的分析来确认它们在评估性能方面与标准UQI的等价性。我们提出了一种新的方法来扩展显微镜的焦深，并使用附加了地面事实的合成图像堆叠，表明该方法优于最先进的方法。我们还证明了作为UQI扩展的度量的输出与UQI的输出是不同的。最后，我们验证了一种新的方法来评估扩展的焦深方法，使用真实世界的切片堆栈，根据UQI，但不需要参考地面真理。Microsc.技术资源。(C)2012年威利期刊公司。
Due to depth of focus constraints, the acquisition of a single 2-D completely in-focus image of 3-D objects characterized by a relevant depth dimension is not possible with a standard light microscope. Since the Seventies numerous methods have been proposed to overcome this problem, mainly through different fusion processing techniques to extend the microscope's depth of focus. However, given a specific application, it is very difficult to know which method yields the best results because there are no validated approaches or tested metrics that are suitable for real world cases typically lacking in a reference ground truth. Although the Universal Quality Index (UQI) is widely used to evaluate output quality in image processing, it requires a reference ground truth. Some UQI extensions have been proposed to evaluate the output of fusion methods without a ground truth, but sufficient analyses have not been carried out to confirm their equivalence to the standard UQI in terms of (evaluation) performance. We propose a new method to extend the microscope's depth of focus and, using synthetic stacks of images with ground truth attached, show that it is superior to state-of-the-art methods. We also demonstrate that the output of metrics proposed as UQI extensions is different from that of the UQI. Finally, we validate a new approach to evaluate extended depth of focus methods using real world stacks of slices, as per the UQI, but without the need for a reference ground truth. Microsc. Res. Tech. (c) 2012 Wiley Periodicals, Inc.