Third Harmonic Generation microscopy distinguishes malignant cell grade in human breast tissue biopsies
Third Harmonic Generation microscopy distinguishes malignant cell grade in human breast tissue biopsies
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DOI：
10.1038/s41598-020-67857-y
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发表时间：
2020-07
影响因子：
4.6
通讯作者：
Evangelia Gavgiotaki;G. Filippidis;Vassilis Tsafas;Savvas Bovasianos;G. Kenanakis;V. Georgoulias;M. Tzardi;S. Agelaki;I. Athanassakis
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文献类型：
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作者：
Evangelia Gavgiotaki;G. Filippidis;Vassilis Tsafas;Savvas Bovasianos;G. Kenanakis;V. Georgoulias;M. Tzardi;S. Agelaki;I. Athanassakis
在外科手术过程中以快速、非侵入性和无染色的方式区分和分级恶性细胞的能力在肿瘤管理中非常重要。为此，应用三次谐波产生（THG）、二次谐波产生（SHG）和傅里叶变换红外（FTIR）光谱来区分人类乳腺组织活检中的恶性细胞与健康细胞。事实上，将非线性过程整合到一个单一的、统一的显微镜平台中，在亚微米水平上提供了单个细胞内的互补结构信息。使用单一激光束，无标记THG成像技术提供了关于平均核和胞质面积、细胞体积和组织强度的重要形态学信息，这些信息在定量时不仅可以区分癌性乳腺组织和良性乳腺组织，而且还可以确定疾病的严重程度。同时，在良性肿瘤组织中，SHG成像可检测到的胶原纤维显示出良好的结构连续性，这些胶原纤维随着疾病的严重程度而沿着逐渐失去方向。THG成像与FTIR光谱的组合可以提供不同级别的乳腺癌之间的更清晰的区分，因为FTIR分析显示恶性组织中的脂质浓度增加。因此，即使在真实的时间外科手术过程中，非线性光学显微镜的使用也可以被认为是用于肿瘤细胞诊断的有力且无害的工具。
The ability to distinguish and grade malignant cells during surgical procedures in a fast, non-invasive and staining-free manner is of high importance in tumor management. To this extend, Third Harmonic Generation (THG), Second Harmonic Generation (SHG) and Fourier-Transform Infrared (FTIR) spectroscopy were applied to discriminate malignant from healthy cells in human breast tissue biopsies. Indeed, integration of non-linear processes into a single, unified microscopy platform offered complementary structural information within individual cells at the submicron level. Using a single laser beam, label-free THG imaging techniques provided important morphological information as to the mean nuclear and cytoplasmic area, cell volume and tissue intensity, which upon quantification could not only distinguish cancerous from benign breast tissues but also define disease severity. Simultaneously, collagen fibers that could be detected by SHG imaging showed a well structured continuity in benign tumor tissues, which were gradually disoriented along with disease severity. Combination of THG imaging with FTIR spectroscopy could provide a clearer distinction among the different grades of breast cancer, since FTIR analysis showed increased lipid concentrations in malignant tissues. Thus, the use of non-linear optical microscopy can be considered as powerful and harmless tool for tumor cell diagnostics even during real time surgery procedures.