Peptide‐PAINT Enables Investigation of Endogenous Talin with Molecular Scale Resolution in Cells and Tissues

Talin is a cell adhesion molecule that is indispensable for the development and function of multicellular organisms. Despite its central role for many cell biological processes, suitable methods to investigate the nanoscale organization of talin in its native environment are missing. Here, we overcome this limitation by combining single‐molecule resolved PAINT (points accumulation in nanoscale topography) imaging with the IRIS (image reconstruction by integrating exchangeable single‐molecule localization) approach, enabling the quantitative analysis of genetically unmodified talin molecules in cells. We demonstrate that a previously reported peptide can be utilized to specifically label the two major talin isoforms expressed in mammalian tissues with a localization precision of
The PIPKIγ‐peptide‐PAINT approach is introduced in this study to enable quantitative investigation of the cell adhesion protein talin with single‐molecule resolution in cells. The technique does not require genetic modifications and allows the analysis of endogenously expressed molecules. The first application reveals atypical adhesion structures in differentiating mesenchymal stem cells.