Bock Nathalie, Röhl Joan
Faculty of Health, School of Biomedical Sciences, Australian Prostate Cancer Research Centre (APCRC-Q), Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), Brisbane, QLD, Australia.
Translational Research Institute (TRI), QUT, Woolloongabba, QLD, Australia.
Methods Mol Biol. 2019;2054:59-77. doi: 10.1007/978-1-4939-9769-5_3.
The study of dynamic processes in the bone metastatic compartment has been challenged by the restrictive access and limited live imaging capabilities that in vivo bone models provide. In this protocol, we show the use of a human bone metastatic bioengineered microtissue for the quantitative investigation of cancer cells in an in vitro bone-like microenvironment. Using live cell epifluorescence microscopy, traditional- and spinning disc-confocal laser scanning microscopy, we demonstrate how to obtain multidimensional real-time data of fluorescently labeled cancer cells in the metastatic microenvironment. Using 4D imaging data processing software such as ImageJ and Imaris, we show how to transform qualitative images and videos into quantitative data of cancer cell attachment, morphology, proliferation, and migration in vitro in the human bone metastatic microtissue.
体内骨模型所提供的有限的观察途径和成像能力，给骨转移微环境中动态过程的研究带来了挑战。在本实验方案中，我们展示了如何使用一种人骨转移性生物工程微组织，在体外类骨微环境中对癌细胞进行定量研究。利用活细胞落射荧光显微镜、传统共聚焦激光扫描显微镜和转盘共聚焦激光扫描显微镜，我们演示了如何在转移微环境中获取荧光标记癌细胞的多维实时数据。利用诸如ImageJ和Imaris等4D成像数据处理软件，我们展示了如何将定性图像和视频转化为体外人骨转移性微组织中癌细胞黏附、形态、增殖和迁移的定量数据。
