Comparing Lifeact and Phalloidin for super-resolution imaging of actin in fixed cells

Visualizing actin filaments in fixed cells is of great interest
for a variety of topics in cell biology such as cell division,
cell movement, and cell signaling. We investigated the
possibility of replacing phalloidin, the standard reagent for
super-resolution imaging of F-actin in fixed cells, with the
actin binding peptide `lifeact’. We compared the labels for use
in single molecule based super-resolution microscopy, where
AlexaFluor 647 labeled phalloidin was used in a (d)STORM modality
and Atto 655 labeled lifeact was used in a single molecule
imaging, reversible binding modality. We found that imaging with
lifeact had a comparable resolution in reconstructed images and
provided several advantages over phalloidin including lower
costs, the ability to image multiple regions of interest on a
coverslip without degradation, simplified
sequential super-resolution imaging, and more continuous labeling
of thin filaments.