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Comparing lifeact and phalloidin for super-resolution imaging of actin in fixed cells.

ABSTRACT: 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.

SUBMITTER: Mazloom-Farsibaf H

PROVIDER: S-EPMC7842966 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Comparing lifeact and phalloidin for super-resolution imaging of actin in fixed cells.

Mazloom-Farsibaf Hanieh H Farzam Farzin F Fazel Mohamadreza M Wester Michael J MJ Meddens Marjolein B M MBM Lidke Keith A KA

PloS one 20210128 1

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 Att ...[more]

PMID: 33508018

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