Total internal reflection fluorescence (TIRF) microscopy of Chlamydomonas flagella
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Authors
Engel, Benjamin D.Lechtreck, Karl-Ferdinand
Sakai, Tsuyoshi
Ikebe, Mitsuo
Witman, George B.
Marshall, Wallace F.
Document Type
Journal ArticlePublication Date
2009-01-01Keywords
AnimalsAxoneme
Biological Transport
Chlamydomonas reinhardtii
Flagella
Fluorescence Recovery After Photobleaching
Green Fluorescent Proteins
Humans
Microscopy, Fluorescence
Molecular Motor Proteins
Protozoan Proteins
Recombinant Fusion Proteins
Cell Biology
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Show full item recordAbstract
The eukaryotic flagellum is host to a variety of dynamic behaviors, including flagellar beating, the motility of glycoproteins in the flagellar membrane, and intraflagellar transport (IFT), the bidirectional traffic of protein particles between the flagellar base and tip. IFT is of particular interest, as it plays integral roles in flagellar length control, cell signaling, development, and human disease. However, our ability to understand dynamic flagellar processes such as IFT is limited in large part by the fidelity with which we can image these behaviors in living cells. This chapter introduces the application of total internal reflection fluorescence (TIRF) microscopy to visualize the flagella of Chlamydomonas reinhardtii. The advantages and challenges of TIRF are discussed in comparison to confocal and differential interference contrast techniques. This chapter also reviews current IFT insights gleaned from TIRF microscopy of Chlamydomonas and provides an outlook on the future of the technique, with particular emphasis on combining TIRF with other emerging imaging technologies.Source
Methods Cell Biol. 2009;93:157-77. Epub 2009 Dec 4. Link to article on publisher's siteDOI
10.1016/S0091-679X(08)93009-0Permanent Link to this Item
http://hdl.handle.net/20.500.14038/51090Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1016/S0091-679X(08)93009-0