Rapid identification of goblet cells in unstained colon thin sections by means of quantum cascade laser-based infrared microspectroscopy
Rapid identification of goblet cells in unstained colon thin sections by means of quantum cascade laser-based infrared microspectroscopy
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DOI：
10.1039/c4an02001d
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发表时间：
2015-01-01
期刊：
影响因子：
4.2
通讯作者：
Petrich, W.
中科院分区：
文献类型：
--
作者：
Kroeger-Lui, N.;Gretz, N.;Petrich, W.
结肠薄片内分泌粘蛋白的杯状细胞区域所覆盖的体积的变化可以作为区分溃疡性结肠炎和感染性结肠炎的手段。在这里，我们表明，基于快速量子级联激光的中红外显微光谱可能有助于溃疡性结肠炎（一种炎症性肠病）的鉴别诊断。每个切片在 7.5 分钟内获得小鼠结肠薄片的红外高光谱图像，像素尺寸为 3.65 x 3.65 μ m(2)，视野为 2.8 x 3.1 mm(2)。通过在一个薄片上的 k 均值聚类的基础上训练随机决策森林分类器来处理光谱。然后将经过训练的算法应用于另外 5 个薄片进行盲法验证，并且能够识别所有切片中的杯状细胞。杯状细胞内糖肽的高含量使得能够快速识别这些未染色、石蜡化的结肠组织薄片中的杯状细胞，正如电磁波谱的 7.6 μm-8.6 μm 和 9.2 μm-9.7 μm 波长范围内明显的光谱特征所揭示的那样。更重要的是，通过简单计算 9.29 μm 和 8.47 μm 处的吸光度值之间的比率，可以将薄片的测量和分析时间进一步缩短至远低于 1 分钟。
Changes in the volume covered by mucin-secreting goblet cell regions within colon thin sections may serve as a means to differentiate between ulcerative colitis and infectious colitis. Here we show that rapid, quantum cascade laser-based mid-infrared micro-spectroscopy might be able to contribute to the differential diagnosis of colitis ulcerosa, an inflammatory bowel disease. Infrared hyperspectral images of mouse colon thin sections were obtained within 7.5 minutes per section with a pixel size of 3.65 x 3.65 mu m(2) and a field of view of 2.8 x 3.1 mm(2). The spectra were processed by training a random decision forest classifier on the basis of k-means clustering on one thin section. The trained algorithm was then applied to 5 further thin sections for a blinded validation and it was able to identify goblet cells in all sections. The rapid identification of goblet cells within these unstained, paraffinized thin sections of colon tissue was enabled by the high content of glycopeptides within the goblet cells as revealed by the pronounced spectral signatures in the 7.6 mu m-8.6 mu m and the 9.2 mu m-9.7 mu m wavelength ranges of the electromagnetic spectrum. More so, the simple calculation of the ratio between the absorbance values at 9.29 mu m and 8.47 mu m provides the potential to further shorten the time for measurement and analysis of a thin section down to well below 1 minute.