Journal
EUROPEAN JOURNAL OF CELL BIOLOGY
Volume 99, Issue 7, Pages -Publisher
ELSEVIER GMBH
DOI: 10.1016/j.ejcb.2020.151106
Keywords
Podosome; Localisation microscopy; Multi-modal microscopy; Super-resolution; AFM
Categories
Funding
- Human Frontier Science Program [RGP0035/2016]
- Royal Society [RG110451]
- MRC [MR/K015664]
- BBSRC [BB/R021767/1]
- Royal Society University Research Fellowship
- Leverhulme Trust Emeritus Fellowship [EM-2018-037]
- BBSRC [BB/R021767/1] Funding Source: UKRI
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Podosomes are mechanosensitive attachment/invasion structures that form on the matrix-adhesion interface of cells and protrude into the extracellular matrix to probe and remodel. Despite their central role in many cellular processes, their exact molecular structure and function remain only partially understood. We review recent progress in molecular scale imaging of podosome architecture, including our newly developed localisation microscopy technique termed HAWK which enables artefact-free live-cell super-resolution microscopy of podosome ring proteins, and report new results on combining fluorescence localisation microscopy (STORM/ PALM) and atomic force microscopy (AFM) on one setup, where localisation microscopy provides the location and dynamics of fluorescently labelled podosome components, while the spatial variation of stiffness is mapped with AFM. For two-colour localisation microscopy we combine iFluor-647, which has previously been shown to eliminate the need to change buffer between imaging modes, with the photoswitchable protein mEOS3.2, which also enables live cell imaging.
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