Single-Molecule Fluorescence Microscopy in Sensory Cilia of Living Caenorhabditis elegans

Elizaveta Loseva; Jaap van Krugten; Aniruddha Mitra; Erwin J.G. Peterman

doi:10.1007/978-1-0716-3377-9_7

Single-Molecule Fluorescence Microscopy in Sensory Cilia of Living Caenorhabditis elegans

Elizaveta Loseva, Jaap van Krugten, Aniruddha Mitra, Erwin J.G. Peterman^*

^*Corresponding author for this work

Research output: Chapter in Book / Report / Conference proceeding › Chapter › Academic › peer-review

Abstract

Intracellular transport of organelles and biomolecules is vital for several cellular processes. Single-molecule fluorescence microscopy can illuminate molecular aspects of the dynamics of individual biomolecules that remain unresolved in ensemble experiments. For example, studying single-molecule trajectories of moving biomolecules can reveal motility properties such as velocity, diffusivity, location and duration of pauses, etc. We use single-molecule imaging to study the dynamics of microtubule-based motor proteins and their cargo in the primary cilia of living C. elegans. To this end, we employ standard fluorescent proteins, an epi-illuminated, widefield fluorescence microscope, and primarily open-source software. This chapter describes the setup we use, the preparation of samples, a protocol for single-molecule imaging in primary cilia of C. elegans, and data analysis.

Original language	English
Title of host publication	Single Molecule Analysis
Subtitle of host publication	Methods and Protocols
Editors	Iddo Heller, David Dulin, Erwin J.G. Peterman
Publisher	Humana Press Inc
Pages	133-150
Number of pages	18
Edition	3rd
ISBN (Electronic)	9781071633779
ISBN (Print)	9781071633762, 9781071633793
DOIs	https://doi.org/10.1007/978-1-0716-3377-9_7
Publication status	Published - 2024

Publication series

Name	Methods in Molecular Biology
Volume	2694
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Funding Information:
We thank Bram Prevo, Jona Mijalkovic, Felix Oswald, and Seyda Acar for their involvement in the earlier stages of this project and discussions. This work is part of the research program of the Foundation for Fundamental Research on Matter (FOM), which is part of the Netherlands Organisation for Scientific Research (NWO). We acknowledge financial support from the European Research Council under the European Union’s Horizon 2020 research and innovation program (Grant agreement no. 788363; “HITSCIL”).

Publisher Copyright:
© The Author(s) 2024.

Keywords

Caenorhabditis elegans
Live-cell imaging
Single-molecule imaging
Widefield fluorescence microscopy

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10.1007/978-1-0716-3377-9_7

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Loseva, E., van Krugten, J., Mitra, A., & Peterman, E. J. G. (2024). Single-Molecule Fluorescence Microscopy in Sensory Cilia of Living Caenorhabditis elegans. In I. Heller, D. Dulin, & E. J. G. Peterman (Eds.), Single Molecule Analysis: Methods and Protocols (3rd ed., pp. 133-150). (Methods in Molecular Biology; Vol. 2694). Humana Press Inc. https://doi.org/10.1007/978-1-0716-3377-9_7

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Single-Molecule Fluorescence Microscopy in Sensory Cilia of Living Caenorhabditis elegans. / Loseva, Elizaveta; van Krugten, Jaap; Mitra, Aniruddha et al.
Single Molecule Analysis: Methods and Protocols . ed. / Iddo Heller; David Dulin; Erwin J.G. Peterman. 3rd. ed. Humana Press Inc, 2024. p. 133-150 (Methods in Molecular Biology; Vol. 2694).

Research output: Chapter in Book / Report / Conference proceeding › Chapter › Academic › peer-review

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Single-Molecule Fluorescence Microscopy in Sensory Cilia of Living Caenorhabditis elegans

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