Protocol

Cell Migration

Volume 769 of the series Methods in Molecular Biology pp 111-136

Date:

Imaging Podosome Dynamics and Matrix Degradation

  • Taylor W. Starnes
  • , Christa L. Cortesio
  • , Anna HuttenlocherAffiliated withDepartment of Medical Microbiology and Immunology, University of Wisconsin-Madison Email author 

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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.

Key words

Podosomes Podosome turnover Podosome dynamics Macrophage THP-1 Live imaging Gelatin degradation Lifeact Invadopodia