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  4. Phalloidin Staining of Actin Filaments for Visualization of Muscle Fibers in Caenorhabditis elegans
 
research article

Phalloidin Staining of Actin Filaments for Visualization of Muscle Fibers in Caenorhabditis elegans

Romani, Mario  
•
Auwerx, Johan  
October 5, 2021
Bio-Protocol

Advances in C. elegans research have allowed scientists to recapitulate different human disorders, from neurodegenerative diseases to muscle dysfunction, in these nematodes. Concomitantly, the interest in visualizing organs affected by these conditions has grown, leading to the establishment of different antibody- and dye-based staining protocols to verify tissue morphology. In particular, the quality of muscle tissue has been largely used in nematodes as a readout for fitness and healthspan. Phalloidin derivatives, which are commonly used to stain actin filaments in cells and tissues, have been implemented in the context of C. elegans research for visualization of muscle fibers. However, the majority of the phalloidin-based protocols depend on fixation steps using harmful compounds, preparation of specific buffers, and large amounts of worms. Herein, we implemented a safer and more flexible experimental procedure to stain actin filaments in C. elegans using phalloidin-based dyes. Lyophilization of the worms followed by their acetone permeabilization allows bypassing the fixation process while also providing the opportunity to suspend the experiment at different steps. Moreover, by using conventional buffers throughout our protocol, we avoid the additional preparation of solutions. Finally, our protocol requires a limited number of worms, making it suitable for slow-growing C. elegans strains. Overall, this protocol provides an efficient, fast, and safer method to stain actin filaments and visualize muscle fibers in C. elegans.

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Type
research article
DOI
10.21769/BioProtoc.4183
Web of Science ID

WOS:000709715100002

Author(s)
Romani, Mario  
Auwerx, Johan  
Date Issued

2021-10-05

Publisher

BIO-PROTOCOL

Published in
Bio-Protocol
Volume

11

Issue

19

Article Number

e4183

Subjects

Biology

•

Life Sciences & Biomedicine - Other Topics

•

phalloidin

•

c. elegans

•

actin

•

muscle fibers

•

cytoskeleton

•

nucleus

•

muscle disease

•

aging

•

c-elegans

•

life-span

•

gene

•

longevity

•

encodes

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LISP  
Available on Infoscience
November 6, 2021
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/182735
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