Ramos-Gomes Fernanda, Ferreira Nathalia, Alves Frauke, Markus M Andrea
Translational Molecular Imaging, Max-Planck-Institute for Multidisciplinary Sciences, Göttingen, Germany.
Clinic of Hematology and Medical Oncology/Institute of Diagnostic and Interventional Radiology, University Medical Center Göttingen, Germany.
Bio Protoc. 2022 Jan 20;12(2):e4293. doi: 10.21769/BioProtoc.4293.
Nanomaterials are increasingly used for the diagnosis and treatment of cancer, including lung cancer. For the clinical translation of nano-based theranostics, it is vital to detect and monitor their accumulation in the tumor, as well as their interaction with tumor, immune cells, and the tumor microenvironment (TME). While high resolution microscopy of fixed tumor specimens can provide some of this information from individual thin slices, it cannot capture cellular events over time and lacks 3D information of the tumor tissue. On the other hand, optical procedures either fall short of providing the necessary cellular resolution, as in the case of epifluorescence optical imaging, or are very demanding, as for instance intravital lung microscopy. We describe an alternative approach to investigate nanoparticle-cell interactions in entire mouse lung lobes, by longitudinal live cell confocal microscopy at nanometer resolution. By filling the lung with 1% agarose, we were able to stabilize the lung lobes and visualize the interaction of fluorescent cells and nanoparticles for at least 4 hours post mortem. This high resolution live cell imaging approach is an easy 4D tool for assessing several dynamic processes in tumor tissue, such as the traffic of cells, shedding of extracellular vesicles (EVs), and the accumulation of nanoparticles in tumor tissue. Graphic abstract: Schematic of the workflow for live cell imaging in the mouse lung.
纳米材料越来越多地用于癌症的诊断和治疗，包括肺癌。对于基于纳米的诊疗方法的临床转化，检测和监测它们在肿瘤中的积累以及它们与肿瘤、免疫细胞和肿瘤微环境（TME）的相互作用至关重要。虽然对固定肿瘤标本进行高分辨率显微镜检查可以从单个薄片中提供一些此类信息，但它无法捕捉随时间变化的细胞事件，并且缺乏肿瘤组织的三维信息。另一方面，光学方法要么无法提供必要的细胞分辨率，如落射荧光光学成像的情况，要么要求非常高，例如活体肺显微镜检查。我们描述了一种通过纳米分辨率的纵向活细胞共聚焦显微镜研究整个小鼠肺叶中纳米颗粒与细胞相互作用的替代方法。通过用1%的琼脂糖填充肺，我们能够稳定肺叶并在死后至少4小时观察荧光细胞和纳米颗粒的相互作用。这种高分辨率活细胞成像方法是一种简单的4D工具，用于评估肿瘤组织中的几个动态过程，如细胞运输、细胞外囊泡（EVs）的脱落以及纳米颗粒在肿瘤组织中的积累。图形摘要：小鼠肺中活细胞成像工作流程示意图。
