Single-molecule fluorescence microscopy in living Caenorhabditis elegans

Jaap van Krugten, Erwin J.G. Peterman*

*Corresponding author for this work

Research output: Chapter in Book / Report / Conference proceedingChapterAcademic

Abstract

Transportation of organelles and biomolecules is vital for many cellular processes. Single-molecule (SM) fluorescence microscopy can expose molecular aspects of the dynamics that remain unresolved in ensemble experiments. For example, trajectories of individual, moving biomolecules can reveal velocity and changes therein, including pauses. We use SM imaging to study the dynamics of motor proteins and their cargo in the cilia of living C. elegans. To this end, we employ standard fluorescent proteins, an epi-illuminated, wide-field fluorescence microscope and mostly open-source software. This chapter describes the setup we use, the preparation of samples, a protocol for single-molecule imaging in C. elegans and data analysis.

Original languageEnglish
Title of host publicationSingle Molecule Analysis
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc
Pages145-154
Number of pages10
ISBN (Electronic)9781493972715
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1665
ISSN (Print)1064-3745

Keywords

  • Caenorhabditis elegans
  • Live-cell imaging
  • Single-molecule imaging
  • Wide-field fluorescence microscopy

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