Zheng Jingtian, Wu Yi-Chien, Cai Xiaoying, Phan Philana, Gill Meghna, Er Ekrem Emrah, Zhao Zongmin, Wang Zaijie J, Lee Steve Seung-Young
Department of Pharmaceutical Sciences, University of Illinois Chicago, Chicago, IL, USA.
Department of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, USA.
iScience. 2025 Feb 3;28(3):111934. doi: 10.1016/j.isci.2025.111934. eCollection 2025 Mar 21.
Three-dimensional (3D) optical microscopy permits interrogation of the tumor microenvironment (TME) in volumetric tumors for research while light sheet and confocal fluorescence microscopy are often used to achieve macroscopic and microscopic 3D images of tissues, respectively. Although each technique offers distinct fields of view (FOVs) and spatial resolution, the combination of the two to obtain correlative multiscale 3D images from the same tumor tissues has not yet been explored. We established a workflow that enables the tracking and 3D imaging of region of interests (ROIs) within tumor tissues through sequential light sheet and confocal fluorescence microscopy. This approach allowed for quantitative 3D spatial analysis of the immune response in the TME at multiple spatial scales and facilitated the direct localization of a metastatic lesion within a mouse brain. Our method offers an approach for correlative multiscale 3D optical microscopy with the potential to provide new insights into comprehensive research in disease mechanism or drug response.
三维（3D）光学显微镜可用于对实体肿瘤中的肿瘤微环境（TME）进行容积式研究，而光片显微镜和共聚焦荧光显微镜通常分别用于获取组织的宏观和微观3D图像。尽管每种技术都提供了不同的视野（FOV）和空间分辨率，但尚未探索将两者结合以从同一肿瘤组织中获得相关的多尺度3D图像。我们建立了一种工作流程，通过顺序光片显微镜和共聚焦荧光显微镜对肿瘤组织内的感兴趣区域（ROI）进行跟踪和3D成像。这种方法允许在多个空间尺度上对TME中的免疫反应进行定量3D空间分析，并有助于在小鼠大脑中直接定位转移灶。我们的方法提供了一种相关多尺度3D光学显微镜方法，有可能为疾病机制或药物反应的综合研究提供新的见解。
