King's College London

Research portal

Combined AFM and super-resolution localisation microscopy: Investigating the structure and dynamics of podosomes

Research output: Contribution to journalArticlepeer-review

Original languageEnglish
Article number151106
JournalEuropean Journal of Cell Biology
Volume99
Issue number7
DOIs
PublishedSep 2020

Documents

  • 200609_invadosome-proc(1)

    200609_invadosome_proc_1_.pdf, 792 KB, application/pdf

    Uploaded date:22 Dec 2020

    Version:Submitted manuscript

King's Authors

Abstract

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.

Download statistics

No data available

View graph of relations

© 2020 King's College London | Strand | London WC2R 2LS | England | United Kingdom | Tel +44 (0)20 7836 5454