title: Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment.
abstract: In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and metastasis. Multicellular interactions in the tumor microenvironment can lead to transient events including directional tumor cell motility and vascular permeability. Quantification of tumor vascular permeability has frequently used end-point experiments to measure extravasation of vascular dyes. However, due to the transient nature of multicellular interactions and vascular permeability, the kinetics of these dynamic events cannot be discerned. By labeling cells and vasculature with injectable dyes or fluorescent proteins, high-resolution time-lapse intravital microscopy has allowed the direct, real-time visualization of transient events in the tumor microenvironment. Here we describe a method for using multiphoton microscopy to perform extended intravital imaging in live mice to directly visualize multicellular dynamics in the tumor microenvironment. This method details cellular labeling strategies, the surgical preparation of a mammary skin flap, the administration of injectable dyes or proteins by tail vein catheter and the acquisition of time-lapse images. The time-lapse sequences obtained from this method facilitate the visualization and quantitation of the kinetics of cellular events of motility and vascular permeability in the tumor microenvironment.
authors.affiliation: Department of Anatomy & Structural Biology, Albert Einstein College of Medicine; Department of Radiology, Albert Einstein College of Medicine; Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine; Integrated Imaging Program, Albert Einstein College of Medicine; allison.harney@einstein.yu.edu.
authors.forename: Allison S
authors.initials: AS
authors.lastname: Harney
authors.affiliation: Department of Anatomy & Structural Biology, Albert Einstein College of Medicine; Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine.
authors.forename: Yarong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Anatomy & Structural Biology, Albert Einstein College of Medicine; Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine; Integrated Imaging Program, Albert Einstein College of Medicine.
authors.forename: John S
authors.initials: JS
authors.lastname: Condeelis
authors.affiliation: Department of Anatomy & Structural Biology, Albert Einstein College of Medicine; Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine; Integrated Imaging Program, Albert Einstein College of Medicine.
authors.forename: David
authors.initials: D
authors.lastname: Entenberg
mesh_terms: D000818:Animals; D002465:Cell Movement; D000069416:Intravital Microscopy; D051379:Mice; D009369:Neoplasms; D009389:Neovascularization, Pathologic; D059016:Tumor Microenvironment