Department of Image Processisng and Biomedical Engineering, Fraunhofer Institute for Integrated Circuits IIS, Erlangen, Germany.
Cytometry A. 2011 Nov;79(11):933-45. doi: 10.1002/cyto.a.21122. Epub 2011 Oct 14.
Interpreting images from fluorescence microscopy is often a time-consuming task with poor reproducibility. Various image processing routines that can help investigators evaluate the images are therefore useful. The critical aspect for a reliable automatic image analysis system is a robust segmentation algorithm that can perform accurate segmentation for different cell types. In this study, several image segmentation methods were therefore compared and evaluated in order to identify the most appropriate segmentation schemes that are usable with little new parameterization and robustly with different types of fluorescence-stained cells for various biological and biomedical tasks. The study investigated, compared, and enhanced four different methods for segmentation of cultured epithelial cells. The maximum-intensity linking (MIL) method, an improved MIL, a watershed method, and an improved watershed method based on morphological reconstruction were used. Three manually annotated datasets consisting of 261, 817, and 1,333 HeLa or L929 cells were used to compare the different algorithms. The comparisons and evaluations showed that the segmentation performance of methods based on the watershed transform was significantly superior to the performance of the MIL method. The results also indicate that using morphological opening by reconstruction can improve the segmentation of cells stained with a marker that exhibits the dotted surface of cells.
荧光显微镜图像的解释通常是一项耗时且重现性差的任务。因此，各种有助于研究人员评估图像的图像处理程序非常有用。对于可靠的自动图像分析系统，关键方面是能够对不同细胞类型进行准确分割的稳健分割算法。在这项研究中，因此比较和评估了几种图像分割方法，以确定最适合的分割方案，这些方案可以在无需大量新参数化的情况下使用，并能够稳健地处理不同类型的荧光染色细胞，用于各种生物学和生物医学任务。本研究调查、比较和增强了用于分离培养上皮细胞的四种不同方法。使用最大强度链接（MIL）方法、改进的 MIL 方法、分水岭方法和基于形态学重建的改进分水岭方法。使用三个手动注释数据集，其中包含 261、817 和 1333 个 HeLa 或 L929 细胞，比较了不同算法。比较和评估表明，基于分水岭变换的方法的分割性能明显优于 MIL 方法。结果还表明，使用重建的形态学开运算可以改善对标记物染色的细胞的分割，该标记物表现出细胞点状表面。
