Le Dévédec Sylvia E, van Roosmalen Wies, Pont Chantal, Lalai Reshma, de Bont Hans, van de Water Bob
Leiden/Amsterdam Center for Drug Research, Division of Toxicology, University of Leiden, Leiden, The Netherlands.
Methods Mol Biol. 2011;769:331-49. doi: 10.1007/978-1-61779-207-6_22.
In the last decade, intravital microscopy on breast tumours in mice at single-cell resolution has resulted in important new insight into mechanisms of metastatic behaviour such as migration, invasion, and intravasation of tumour cells; angiogenesis; and the response of immune cells. This chapter describes the methods that can be used for analysing tumour cell motility in a mouse model of breast cancer metastasis. It includes protocols for generation of a labelled primary tumour, its imaging with two-photon microscopy, and the processing of time-lapse image data. Furthermore, we present a methodology, recently developed in our laboratory that combines multicolour imaging with an inducible cell model to study the role of a specific gene of interest in tumour cell motility in vivo. This protocol can be used to image the metastatic behaviour of different individual tumour cells within the same tumour microenvironment and correlate it with metastasis formation. Additional protocols for labelling macrophages to visualise blood flow and image analysis are also included.
在过去十年中，以单细胞分辨率对小鼠乳腺肿瘤进行的活体显微镜检查，已为转移行为机制带来了重要的新见解，如肿瘤细胞的迁移、侵袭和血管内渗；血管生成；以及免疫细胞的反应。本章描述了可用于在乳腺癌转移小鼠模型中分析肿瘤细胞运动性的方法。它包括生成标记原发性肿瘤的方案、用双光子显微镜对其成像以及延时图像数据的处理。此外，我们还介绍了一种最近在我们实验室开发的方法，该方法将多色成像与可诱导细胞模型相结合，以研究体内特定感兴趣基因在肿瘤细胞运动性中的作用。该方案可用于在同一肿瘤微环境中对不同的单个肿瘤细胞的转移行为进行成像，并将其与转移形成相关联。还包括标记巨噬细胞以可视化血流和图像分析的其他方案。
