Two-Photon Intravital Multicolour Imaging to Study Metastatic Behaviour of Cancer Cells In Vivo
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.
- In Vivo Tumor Formation From Primate Embryonic Stem Cells
- Detection and Quantitation of mRNAs Using Ribonuclease Protection Assays
- Optimizing Sporulation Conditions for Different Saccharomyces cerevisiae Strain Backgrounds
- Retroviral Transduction of Murine Primary T Lymphocytes
- Recording of Ion Channel Activity in Planar Lipid Bilayer Experiments
- SpermEgg Fusion Assay in Mammals
- T-Cell Activation Using mAb to CD3
- Isolation, Culture, and Characterization of Human Umbilical Cord Stroma-derived Mesenchymal Stem Cells
- High-Throughput RNAi Screen in Drosophila
- Cell Cycle Staining Protocol-DAPI