Spectral histopathology of colon cancer tissue sections by Raman imaging with 532 nm excitation provides label free annotation of lymphocytes, erythrocytes and proliferating nuclei of cancer cells
Spectral histopathology of colon cancer tissue sections by Raman imaging with 532 nm excitation provides label free annotation of lymphocytes, erythrocytes and proliferating nuclei of cancer cells
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DOI：
10.1039/c3an00370a
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发表时间：
2013-01-01
期刊：
影响因子：
4.2
通讯作者：
Gerwert, Klaus
中科院分区：
文献类型：
--
作者：
Mavarani, Laven;Petersen, Dennis;Gerwert, Klaus
光谱组织病理学（SHP）是一种新兴的无标记组织注释工具。虽然基于FTIR的SHP提供较大组织切片的快速注释，但是基于拉曼的SHP较慢，但是与FTIR相比实现了10倍高的空间分辨率。通常NIR激发用于生物样品的拉曼测量。这里，首次使用532 nm激发通过拉曼SHP来注释结肠组织。获得了出色的数据质量，例如红细胞和淋巴细胞。除拉曼散射外，还观察到自发荧光。我们发现，这种自体荧光在空间上与手术病理学中常规免疫组织化学中使用的抗p53抗体的荧光重叠。这种荧光指示具有突变的p53的癌细胞的细胞核，并允许癌细胞的新的无标记分配。这些结果为利用拉曼光谱和自发荧光进行光学诊断开辟了新的途径。
Spectral histopathology (SHP) is an emerging tool for label free annotation of tissue. While FTIR based SHP provides fast annotation of larger tissue sections, Raman based SHP is slower but achieves a 10 times higher spatial resolution as compared to FTIR. Usually NIR excitation is used for Raman measurements on biological samples. Here, for the first time 532 nm excitation is used to annotate colon tissue by Raman SHP. Excellent data quality is obtained, which resolves for example erythrocytes and lymphocytes. In addition to Raman scattering auto-fluorescence is observed. We found that this auto-fluorescence overlaps spatially with the fluorescence of antibodies against p53 used in routine immunohistochemistry in surgical pathology. This fluorescence indicates nuclei of cancer cells with mutated p53 and allows new label free assignment of cancer cells. These results open new avenues for optical diagnosis by Raman spectroscopy and autofluorescence.