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...

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Bibliographic Details
Published in:PloS one 2021-01, Vol.16 (1), p.e0246138
Main Authors: Mazloom-Farsibaf, Hanieh, Farzam, Farzin, Fazel, Mohamadreza, Wester, Michael J, Meddens, Marjolein B M, Lidke, Keith A
Format: Article
Language:English
Subjects:
Actin
Actin Cytoskeleton - chemistry
Actin Cytoskeleton - pathology
Astronomy
Basophilic leukemia
Biology and Life Sciences
Carbocyanines - chemistry
Carbon dioxide
Cell adhesion & migration
Cell research
Chemical tests and reagents
Comparative analysis
Composition
Cytoskeleton
Deoxyribonucleic acid
Diffraction patterns
DNA
Glucose
Glutamine
Health care facilities
HeLa Cells
Humans
Image resolution
Labeling
Leukemia
Light diffraction
Light microscopy
Methods
Microscope and microscopy
Microscopy, Fluorescence
Nuclear medicine
Observations
Optical microscopy
Penicillin
Peptides
Phalloidin
Phalloidine - chemistry
Physics
Protein binding
Proteins
Radiology
Research and analysis methods
Stochasticity
Streptomycin
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0246138