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Multimodal widefield fluorescence imaging with nonlinear optical microscopy workflow for noninvasive oral epithelial neoplasia detection: a preclinical study
Journal of Biomedical Optics ( IF 3.5 ) Pub Date : 2020-11-01 , DOI: 10.1117/1.jbo.25.11.116008
Rahul Pal 1, 2 , Paula Villarreal 3 , Xiaoying Yu 3 , Suimin Qiu 3 , Gracie Vargas 3
Affiliation  

Significance: Early detection of epithelial cancers and precancers/neoplasia in the presence of benign lesions is challenging due to the lack of robust in vivo imaging and biopsy guidance techniques. Label-free nonlinear optical microscopy (NLOM) has shown promise for optical biopsy through the detection of cellular and extracellular signatures of neoplasia. Although in vivo microscopy techniques continue to be developed, the surface area imaged in microscopy is limited by the field of view. FDA-approved widefield fluorescence (WF) imaging systems that capture autofluorescence signatures of neoplasia provide molecular information at large fields of view, which may complement the cytologic and architectural information provided by NLOM. Aim: A multimodal imaging approach with high-sensitivity WF and high-resolution NLOM was investigated to identify and distinguish image-based features of neoplasia from normal and benign lesions. Approach:In vivo label-free WF imaging and NLOM was performed in preclinical hamster models of oral neoplasia and inflammation. Analyses of WF imaging, NLOM imaging, and dual modality (WF combined with NLOM) were performed. Results: WF imaging showed increased red-to-green autofluorescence ratio in neoplasia compared to inflammation and normal oral mucosa (p < 0.01). In vivo assessment of the mucosal tissue with NLOM revealed subsurface cytologic (nuclear pleomorphism) and architectural (remodeling of extracellular matrix) atypia in histologically confirmed neoplastic tissue, which were not observed in inflammation or normal mucosa. Univariate and multivariate statistical analysis of macroscopic and microscopic image-based features indicated improved performance (94% sensitivity and 97% specificity) of a multiscale approach over WF alone, even in the presence of benign lesions (inflammation), a common confounding factor in diagnostics. Conclusions: A multimodal imaging approach integrating strengths from WF and NLOM may be beneficial in identifying oral neoplasia. Our study could guide future studies on human oral neoplasia to further evaluate merits and limitations of multimodal workflows and inform the development of multiscale clinical imaging systems.

中文翻译:

用于无创口腔上皮肿瘤检测的多模态宽场荧光成像与非线性光学显微镜工作流程:临床前研究

意义:由于缺乏强大的体内成像和活检引导技术,在存在良性病变的情况下早期检测上皮癌和癌前期/肿瘤形成具有挑战性。无标记非线性光学显微镜(NLOM)通过检测肿瘤的细胞和细胞外特征,显示出光学活检的前景。尽管体内显微镜技术不断发展,但显微镜成像的表面积受到视野的限制。FDA 批准的宽场荧光 (WF) 成像系统可捕获肿瘤的自发荧光特征,提供大视野的分子信息,这可以补充 NLOM 提供的细胞学和结构信息。目的:研究采用高灵敏度 WF 和高分辨率 NLOM 的多模态成像方法,以识别和区分肿瘤的基于图像的特征与正常和良性病变。方法:在口腔肿瘤和炎症的临床前仓鼠模型中进行体内无标记 WF 成像和 NLOM。对 WF 成像、NLOM 成像和双模态(WF 与 NLOM 结合)进行了分析。结果:WF 成像显示,与炎症和正常口腔粘膜相比,肿瘤中红绿自发荧光比率增加 (p < 0.01)。用 NLOM 对粘膜组织进行体内评估,揭示了组织学证实的肿瘤组织中的表面下细胞学(核多形性)和结构(细胞外基质的重塑)异型性,而在炎症或正常粘膜中未观察到。基于宏观和微观图像特征的单变量和多变量统计分析表明,即使存在良性病变(炎症)(诊断中常见的混杂因素),多尺度方法的性能也优于单独的 WF(94% 敏感性和 97% 特异性) 。结论:结合 WF 和 NLOM 优势的多模态成像方法可能有助于识别口腔肿瘤。我们的研究可以指导未来对人类口腔肿瘤的研究,以进一步评估多模式工作流程的优点和局限性,并为多尺度临床成像系统的开发提供信息。
更新日期:2020-11-16
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