Protocol for Determining the Average Speed and Frequency of Kinesin and Dynein-Driven Intraflagellar Transport (IFT) in C. Elegans

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

doi:10.1016/j.xpro.2022.101498

Protocol for Determining the Average Speed and Frequency of Kinesin and Dynein-Driven Intraflagellar Transport (IFT) in C. Elegans

Abstract

Here, we present a protocol to image a fluorescent-labeled intraflagellar trans-port (IFT) component in Caenorhabditis elegans with fluorescence microscopy, including steps of sample preparations, in vivo live-cell imaging, and post -micro-scopy 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).

Description

Cakici, Onur/0000-0002-6809-4467; Kaplan, Oktay Ismail/0000-0002-8733-0920; Turan, Merve Gul/0000-0001-5783-7168

ORCID

Keywords

Molecular Biology, Cell Biology, Genetics, Model Organisms, Neuroscience, Bioinformatics, Microscopy, CRISPR, Microscopy, Science (General), Bioinformatics, Dyneins, Kinesins, Cell Biology, Q1-390, #cilia #IFT #celegans, Model Organisms, Flagella, CRISPR, Mutation, Genetics, Protocol, Animals, Cilia, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Molecular Biology, Neuroscience