Deverell M H, Salisbury J R, Whimster W F
Department of Morbid Anatomy, King's College Hospital, London, UK.
Pathol Res Pract. 1989 Nov;185(5):555-7. doi: 10.1016/S0344-0338(89)80192-X.
Nuclear measurements using image analysis largely depend upon the quality of the image presented for digitization. To investigate the effects of nuclear stains on image segmentation of nuclei, serial sections of kidney were stained by eleven different methods and presented to an IBAS 2000 interactive image analysis system (Kontron Bildanalyse) via a Zeiss IIIRS microscope at x 800 magnification and a Siemens K30 video camera. Digitised grey level images of each field were processed by an interactive technique and by an automatic segmentation procedure (thresholding). Nuclear areas were measured by each method and the results compared. We conclude that of the stains assessed the uncounterstained haematoxylins offer the best image segmentation for nuclear measurements. Thresholding techniques are suitable for performing measurements using these stains, particularly when additional interactive techniques are used to reject unwanted structures and to separate overlapping nuclei after segmentation. Comparable areas stained with five of the stains were studied to see if the staining techniques themselves affected nuclear area. Our results show that the use of different stains will substantially affect measurements of nuclear dimensions.
使用图像分析进行细胞核测量在很大程度上取决于用于数字化的图像质量。为了研究细胞核染色剂对细胞核图像分割的影响，采用11种不同方法对肾脏连续切片进行染色，并通过蔡司IIIRS显微镜（放大倍数为800倍）和西门子K30摄像机将其呈现给IBAS 2000交互式图像分析系统（康强图像分析公司）。通过交互式技术和自动分割程序（阈值处理）对每个视野的数字化灰度图像进行处理。用每种方法测量细胞核面积并比较结果。我们得出结论，在所评估的染色剂中，未复染的苏木精为细胞核测量提供了最佳的图像分割效果。阈值处理技术适用于使用这些染色剂进行测量，特别是当使用额外的交互式技术来排除不需要的结构并在分割后分离重叠的细胞核时。对用其中五种染色剂染色的可比区域进行了研究，以观察染色技术本身是否会影响细胞核面积。我们的结果表明，使用不同的染色剂会显著影响细胞核尺寸的测量。
