Total internal reflection fluorescence (TIRF) microscopy of Chlamydomonas flagella
Engel, Benjamin D. ; Lechtreck, Karl-Ferdinand ; Sakai, Tsuyoshi ; Ikebe, Mitsuo ; Witman, George B. ; Marshall, Wallace F.
Citations
Authors
Lechtreck, Karl-Ferdinand
Sakai, Tsuyoshi
Ikebe, Mitsuo
Witman, George B.
Marshall, Wallace F.
Student Authors
Faculty Advisor
Academic Program
UMass Chan Affiliations
Document Type
Publication Date
Subject Area
Collections
Embargo Expiration Date
Link to Full Text
Abstract
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 site