Abstract
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.
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Acknowledgements
We thank J. Wyckoff for helpful suggestions for our intravital imaging work, M. Balzar for helpful advises concerning the multiphoton microscope, and L. Price for critically reading the manuscript. This work was financially supported by grants from the Dutch Cancer Society (UL 2007-3860), the EU FP7 Health Program Metafight (Grant agreement no.201862), and the Netherlands Organization for Scientific Research (902-21-229 and 911-02-022).
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Le Dévédec, S.E., van Roosmalen, W., Pont, C., Lalai, R., de Bont, H., van de Water, B. (2011). Two-Photon Intravital Multicolour Imaging to Study Metastatic Behaviour of Cancer Cells In Vivo. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology, vol 769. Humana Press. https://doi.org/10.1007/978-1-61779-207-6_22
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DOI: https://doi.org/10.1007/978-1-61779-207-6_22
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