Single-molecule fluorescence microscopy in living Caenorhabditis elegans

Jaap van Krugten; Erwin J.G. Peterman

doi:10.1007/978-1-4939-7271-5_8

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 proceeding › Chapter › Academic

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 language	English
Title of host publication	Single Molecule Analysis
Subtitle of host publication	Methods and Protocols
Editors	Erwin J.G. Peterman
Publisher	Humana Press Inc
Pages	145-154
Number of pages	10
ISBN (Electronic)	9781493972715
DOIs	https://doi.org/10.1007/978-1-4939-7271-5_8
Publication status	Published - 2018

Publication series

Name	Methods in Molecular Biology
Volume	1665
ISSN (Print)	1064-3745

Keywords

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

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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.",

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AB - 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.

KW - Caenorhabditis elegans

KW - Live-cell imaging

KW - Single-molecule imaging

KW - Wide-field fluorescence microscopy

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ER -

Single-molecule fluorescence microscopy in living Caenorhabditis elegans

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