TY - JOUR
T1 - Comparing lifeact and phalloidin for super-resolution imaging of actin in fixed cells
AU - Mazloom-Farsibaf, Hanieh
AU - Farzam, Farzin
AU - Fazel, Mohamadreza
AU - Wester, Michael J
AU - Meddens, Marjolein B M
AU - Lidke, Keith A
N1 - Publisher Copyright:
Copyright: © 2021 Mazloom-Farsibaf et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2021/1/28
Y1 - 2021/1/28
N2 - 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 dSTORM 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.
AB - 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 dSTORM 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.
KW - Actin Cytoskeleton/chemistry
KW - Carbocyanines/chemistry
KW - HeLa Cells
KW - Humans
KW - Microscopy, Fluorescence
KW - Phalloidine/chemistry
U2 - 10.1371/journal.pone.0246138
DO - 10.1371/journal.pone.0246138
M3 - Article
C2 - 33508018
SN - 1932-6203
VL - 16
JO - PLoS ONE
JF - PLoS ONE
IS - 1 January
M1 - e0246138
ER -