title: Imaging podosome dynamics and matrix degradation.
abstract: Invasive cell migration is critical for leukocyte trafficking into tissues. Podosomes are matrix-degrading adhesive structures that are formed by macrophages and are necessary for macrophage migration and invasion. Here, we describe methods for imaging and quantifying podosomes in primary human macrophages and in THP-1 cells, a monocyte cell line that can be differentiated to a macrophage-like state. Moreover, we outline detailed methods for live imaging of podosomes, which are highly dynamic, and for the quantification of rates of podosome turnover. Finally, we discuss methods for the quantitative analysis of matrix degradation on fluorescent-gelatin-coated cover slips.
authors: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI, USA.
authors: Taylor W
authors: TW
authors: Starnes
authors: Christa L
authors: CL
authors: Cortesio
authors: Anna
authors: A
authors: Huttenlocher
mesh_terms: D002454:Cell Differentiation; D002460:Cell Line; D021982:Cell-Matrix Junctions / Q000378:metabolism*; D002633:Chemotaxis; D020099:Coated Materials, Biocompatible; D005109:Extracellular Matrix / Q000378:metabolism*; D022001:Focal Adhesions / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000502:physiology; D008853:Microscopy / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods*; D059008:Time-Lapse Imaging; D016596:Vinculin / Q000096:biosynthesis / Q000235:genetics
chemical_list: D020099:Coated Materials, Biocompatible; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D016596:Vinculin; D049452:Green Fluorescent Proteins; D005780:Gelatin