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首页> 美国卫生研究院文献>Journal of Biomedical Optics >Confocal mosaicing microscopy of human skin ex vivo: spectral analysis for digital staining to simulate histology-like appearance
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Confocal mosaicing microscopy of human skin ex vivo: spectral analysis for digital staining to simulate histology-like appearance

机译：离体人体皮肤的共聚焦镶嵌显微镜：用于数字染色的光谱分析以模拟组织学样外观

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Confocal mosaicing microscopy enables rapid imaging of large areas of fresh tissue, without the processing that is necessary for conventional histology. Mosaicing may offer a means to perform rapid histology at the bedside. A possible barrier toward clinical acceptance is that the mosaics are based on a single mode of grayscale contrast and appear black and white, whereas histology is based on two stains (hematoxylin for nuclei, eosin for cellular cytoplasm and dermis) and appears purple and pink. Toward addressing this barrier, we report advances in digital staining: fluorescence mosaics that show only nuclei, are digitally stained purple and overlaid on reflectance mosaics, which show only cellular cytoplasm and dermis, and are digitally stained pink. With digital staining, the appearance of confocal mosaics mimics the appearance of histology. Using multispectral analysis and color matching functions, red, green, and blue (RGB) components of hematoxylin and eosin stains in tissue were determined. The resulting RGB components were then applied in a linear algorithm to transform fluorescence and reflectance contrast in confocal mosaics to the absorbance contrast seen in pathology. Optimization of staining with acridine orange showed improved quality of digitally stained mosaics, with good correlation to the corresponding histology.
机译：共聚焦镶嵌显微镜可以对新鲜组织的大面积进行快速成像，而无需进行常规组织学所需的处理。镶嵌术可能提供一种在床边进行快速组织学检查的方法。临床接受的可能障碍是马赛克基于单一的灰度对比模式并显示为黑色和白色，而组织学基于两种染色（苏木素用于细胞核，曙红用于细胞质和真皮）并且呈现紫色和粉红色。为了解决这一障碍，我们报告了数字染色的进展：仅显示细胞核的荧光马赛克已被数字染色为紫色，并覆盖了反射马赛克，仅显示了细胞质和真皮，并且被数字染色为粉红色。通过数字染色，共焦马赛克的外观模仿了组织学的外观。使用多光谱分析和颜色匹配功能，确定组织中苏木精和曙红染色的红色，绿色和蓝色（RGB）成分。然后将所得的RGB分量应用到线性算法中，以将共焦马赛克中的荧光和反射率对比度转换为病理学中看到的吸收率对比度。用a啶橙染色的优化表明数字染色的镶嵌物质量提高，与相应的组织学有很好的相关性。

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