title: Real-time imaging of myeloid cells dynamics in ApcMin/+ intestinal tumors by spinning disk confocal microscopy.
abstract: Myeloid cells are the most abundant immune cells within tumors and have been shown to promote tumor progression. Modern intravital imaging techniques enable the observation of live cellular behavior inside the organ but can be challenging in some types of cancer due to organ and tumor accessibility such as intestine. Direct observation of intestinal tumors has not been previously reported. A surgical procedure described here allows direct observation of myeloid cell dynamics within the intestinal tumors in live mice by using transgenic fluorescent reporter mice and injectable tracers or antibodies. For this purpose, a four-color, multi-region, micro-lensed spinning disk confocal microscope that allows long-term continuous imaging with rapid image acquisition has been used. Apc(Min/+) mice that develop multiple adenomas in the small intestine are crossed with c-fms-EGFP mice to visualize myeloid cells and with ACTB-ECFP mice to visualize intestinal epithelial cells of the crypts. Procedures for labeling different tumor components, such as blood vessels and neutrophils, and the procedure for positioning the tumor for imaging through the serosal surface are also described. Time-lapse movies compiled from several hours of imaging allow the analysis of myeloid cell behavior in situ in the intestinal microenvironment.
authors.affiliation: Department of Oncology, INSERM U661, Functional Genomic Institute.|Department of Anatomy, University of California.
authors.forename: Caroline
authors.initials: C
authors.lastname: Bonnans
authors.affiliation: Department of Anatomy, University of California.
authors.forename: Marja
authors.initials: M
authors.lastname: Lohela
authors.affiliation: Department of Anatomy, University of California.
authors.forename: Zena
authors.initials: Z
authors.lastname: Werb
mesh_terms: D000236:Adenoma / Q000473:pathology*; D000818:Animals; D003199:Computer Systems; D007414:Intestinal Neoplasms / Q000473:pathology*; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000379:methods*; D022423:Myeloid Cells / Q000473:pathology*