Basic Methods to Visualize Actin Filaments In Vitro Using Fluorescence Microscopy for Observation of Filament Severing and Bundling.
Dynamics of actin filaments are regulated by a number of actin-binding proteins. To understand the function of an actin-binding protein, it is necessary to characterize effects of the protein on actin filament dynamics in vitro. This chapter describes basic microscopic methods to visualize fluorescently labeled actin filaments using commonly available fluorescence microscope settings. Direct microscopic observation of actin filaments provides strong evidence for severing or bundling of actin filaments.
Department of Pathology, Emory University, 615 Michael Street, Whitehead Research Building, Room 105N, Atlanta, GA, 30322, USA. sono@emory.edu.
Shoichiro
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D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000737:chemistry; D000818:Animals; D003101:Collodion / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods*; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D011817:Rabbits
D000199:Actins; D005456:Fluorescent Dyes; D003101:Collodion
Actin filament dynamics; Actin-binding proteins; Direct observation; Fluorescence microscopy; Fluorophore labeling