Department of Biophysics and Protein Research Unit Europe (PURE), Ruhr University Bochum, ND/04 Nord, 44780 Bochum, Germany.
Analyst. 2013 Jul 21;138(14):4035-9. doi: 10.1039/c3an00370a. Epub 2013 Jun 4.
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.
光谱组织病理学（SHP）是一种用于组织无标记注释的新兴工具。虽然基于傅里叶变换红外光谱（FTIR）的 SHP 可快速注释较大的组织切片，但基于拉曼光谱（Raman）的 SHP 速度较慢，但与 FTIR 相比，其空间分辨率要高 10 倍。通常，近红外（NIR）激发用于对生物样本进行拉曼测量。在这里，首次使用 532nm 激发来通过拉曼 SHP 注释结肠组织。获得了出色的数据质量，可以分辨例如红细胞和淋巴细胞。此外，还观察到拉曼散射自荧光。我们发现这种自荧光在空间上与用于外科病理学常规免疫组织化学的 p53 抗体的荧光重叠。这种荧光表明了突变型 p53 的癌细胞的细胞核，并允许对癌细胞进行新的无标记分配。这些结果为通过拉曼光谱和自发荧光进行光学诊断开辟了新途径。
