title: Two-photon intravital multicolour imaging to study metastatic behaviour of cancer cells in vivo.
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.
authors: Leiden/Amsterdam Center for Drug Research, Division of Toxicology, University of Leiden, Leiden, The Netherlands.
authors: Sylvia E
authors: SE
authors: Le Dévédec
authors: Wies
authors: W
authors: van Roosmalen
authors: Chantal
authors: C
authors: Pont
authors: Reshma
authors: R
authors: Lalai
authors: Hans
authors: H
authors: de Bont
authors: Bob
authors: B
authors: van de Water
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002465:Cell Movement; D058948:Cell Tracking / Q000295:instrumentation / Q000379:methods; D005260:Female; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008175:Lung Neoplasms / Q000378:metabolism / Q000556:secondary; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D036641:Microscopy, Fluorescence, Multiphoton; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D009362:Neoplasm Metastasis; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013194:Staining and Labeling; D059008:Time-Lapse Imaging
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins