STAR Protocols (Sep 2022)

Protocol for determining the average speed and frequency of kinesin and dynein-driven intraflagellar transport (IFT) in C. elegans

  • Merve G. Turan,
  • Hanife Kantarci,
  • Sadiye D. Temtek,
  • Onur Cakici,
  • Sebiha Cevik,
  • Oktay I. Kaplan

Journal volume & issue
Vol. 3, no. 3
p. 101498

Abstract

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Summary: Here, we present a protocol to image a fluorescent-labeled intraflagellar transport (IFT) component in Caenorhabditis elegans with fluorescence microscopy, including steps of sample preparations, in vivo live-cell imaging, and post-microscopy analysis with kymographs. This protocol breaks down all processes into three categories: (1) pre-imaging preparations, (2) preparations for the time of image acquisition, and (3) post-imaging analyses. The protocol can be applied to determine the speed and frequency of moving particles.For complete details on the use and execution of this protocol, please refer to Cevik et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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