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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| Veröffentlicht in: | PloS one 2021-01, Vol.16 (1), p.e0246138 |
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| creator | Mazloom-Farsibaf, Hanieh Farzam, Farzin Fazel, Mohamadreza Wester, Michael J Meddens, Marjolein B M Lidke, Keith A |
| description | 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. |
| doi_str_mv | 10.1371/journal.pone.0246138 |
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This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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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. 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| 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 |
| title | Comparing lifeact and phalloidin for super-resolution imaging of actin in fixed cells |
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