Villarreal Paula Patricia, Pal Rahul, Qiu Suimin, Coblens Orly, Villasante-Tezanos Alejandro, Resto Vicente, McCammon Susan, Vargas Gracie
The Institute for Translational Sciences, University of Texas Medical Branch, Galveston, TX 77555, USA.
Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Charlestown, MA 02129, USA.
Cancers (Basel). 2023 Feb 18;15(4):1302. doi: 10.3390/cancers15041302.
Depth-resolved label-free optical imaging by the method of multiphoton autofluorescence microscopy (MPAM) may offer new ways to examine cellular and extracellular atypia associated with epithelial squamous cell carcinoma (SCC). MPAM was evaluated for its ability to identify cellular and microstructural atypia in head and neck tissues from resected discarded tumor tissue. Three-dimensional image volumes were obtained from tissues from the floor of the mouth, tongue, and larynx, and were then processed for histology. MPAM micrographs were evaluated for qualitative metrics of cell atypia and quantitative measures associated with nuclear pleomorphism. Statistical analyses correlated MPAM endpoints with histological grade from each imaged site. Cellular overcrowding, discohesion, anisonucleosis, and multinucleated cells, as observed through MPAM, were found to be statistically associated with dysplasia and SCC grading, but not in histologically benign regions. A quantitative measure of the coefficient of variance in nuclear size in SCC and dysplasia was statistically elevated above histologically benign regions. MPAM also allowed for the identification of cellular heterogeneity across transitional areas and other features, such as inflammatory infiltrates. In the future, MPAM could be evaluated for the non-invasive detection of neoplasia, possibly as an adjunct to traditional conventional examination and biopsy.
通过多光子自发荧光显微镜（MPAM）方法进行深度分辨无标记光学成像，可能为检查与上皮鳞状细胞癌（SCC）相关的细胞和细胞外异型性提供新途径。对MPAM识别来自切除废弃肿瘤组织的头颈部组织中细胞和微观结构异型性的能力进行了评估。从口腔底部、舌头和喉部的组织中获取三维图像体积，然后进行组织学处理。对MPAM显微照片进行细胞异型性的定性指标和与核多形性相关的定量测量评估。统计分析将MPAM终点与每个成像部位的组织学分级相关联。通过MPAM观察到的细胞过度拥挤、细胞间离散、核大小不一和多核细胞，在统计学上与发育异常和SCC分级相关，但在组织学上良性区域则不然。SCC和发育异常中核大小方差系数的定量测量在统计学上高于组织学上良性区域。MPAM还能够识别过渡区域的细胞异质性以及其他特征，如炎性浸润。未来，可以评估MPAM用于肿瘤的非侵入性检测，可能作为传统常规检查和活检的辅助手段。
