Chapter title |
Single-Molecule Fluorescence Microscopy in Sensory Cilia of Living Caenorhabditis elegans
|
---|---|
Chapter number | 7 |
Book title |
Single Molecule Analysis
|
Published by |
Humana, New York, NY, October 2023
|
DOI | 10.1007/978-1-0716-3377-9_7 |
Pubmed ID | |
Book ISBNs |
978-1-07-163376-2, 978-1-07-163377-9
|
Authors |
Elizaveta Loseva, Jaap van Krugten, Aniruddha Mitra, Erwin J G Peterman |
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. |
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Geographical breakdown
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