title: Quantification of transendothelial migration using three-dimensional confocal microscopy.
abstract: Migration of cells across endothelial barriers, termed transendothelial migration (TEM), is an important cellular process that underpins the pathology of many disease states including chronic inflammation and cancer metastasis. While this process can be modeled in vitro using cultured cells, many model systems are unable to provide detailed visual information of cell morphologies and distribution of proteins such as junctional markers, as well as quantitative data on the rate of TEM. Improvements in imaging techniques have made microscopy-based assays an invaluable tool for studying this type of detailed cell movement in physiological processes. In this chapter, we describe a confocal microscopy-based method that can be used to assess TEM of both leukocytes and cancer cells across endothelial barriers in response to a chemotactic gradient, as well as providing information on their migration into a subendothelial extracellular matrix, designed to mimic that found in vivo.
authors: Cell Motility and Cytoskeleton Group, Randall Division of Cell and Molecular Biophysics, King’s College London, London, UK.
authors: Robert J
authors: RJ
authors: Cain
authors: Bárbara Borda
authors: BB
authors: d'Água
authors: Anne J
authors: AJ
authors: Ridley
mesh_terms: D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D054443:Cell Migration Assays / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques; D024042:Collagen Type I / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods*; D057705:Transendothelial and Transepithelial Migration; D014584:User-Computer Interface
chemical_list: D024042:Collagen Type I