English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Complementarity of PALM and SOFI for super-resolution live-cell imaging of focal adhesions.

MPS-Authors
/persons/resource/persons15441

Leutenegger,  M.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

External Resource
No external resources are shared
Fulltext (restricted access)
There are currently no full texts shared for your IP range.
Fulltext (public)

2378504.pdf
(Publisher version), 2MB

Supplementary Material (public)

2378504_Suppl_1.pdf
(Supplementary material), 568KB

2378504_Suppl_2.pdf
(Supplementary material), 25MB

2378504_Suppl_3.avi
(Supplementary material), 3MB

Citation

Deschout, H., Lukes, T., Sharipov, A., Szlag, D., Feletti, L., Vandenberg, W., et al. (2016). Complementarity of PALM and SOFI for super-resolution live-cell imaging of focal adhesions. Nature Communications, 7: 13693. doi:10.1038/ncomms13693.


Cite as: https://hdl.handle.net/11858/00-001M-0000-002C-31A5-7
Abstract
Live-cell imaging of focal adhesions requires a sufficiently high temporal resolution, which remains a challenge for super-resolution microscopy. Here we address this important issue by combining photoactivated localization microscopy (PALM) with super-resolution optical fluctuation imaging (SOFI). Using simulations and fixed-cell focal adhesion images, we investigate the complementarity between PALM and SOFI in terms of spatial and temporal resolution. This PALM-SOFI framework is used to image focal adhesions in living cells, while obtaining a temporal resolution below 10 s. We visualize the dynamics of focal adhesions, and reveal local mean velocities around 190 nmmin (-1). The complementarity of PALM and SOFI is assessed in detail with a methodology that integrates a resolution and signal-to-noise metric. This PALM and SOFI concept provides an enlarged quantitative imaging framework, allowing unprecedented functional exploration of focal adhesions through the estimation of molecular parameters such as fluorophore densities and photoactivation or photoswitching kinetics.