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Vol. 10, Issue 1, 197-210, January 1999

Birefringence Imaging Directly Reveals Architectural Dynamics of Filamentous Actin in Living Growth Cones

Kaoru Katoh, Katherine Hammar, Peter J. S. Smith, and Rudolf Oldenbourg^*

Marine Biological Laboratory, Woods Hole, Massachusetts 02543-1015

We have investigated the dynamic behavior of cytoskeletal fine structure in the lamellipodium of nerve growth cones using a new type of polarized light microscope (the Pol-Scope). Pol-Scope images display with exquisite resolution and definition birefringent fine structures, such as filaments and membranes, without having to treat the cell with exogenous dyes or fluorescent labels. Furthermore, the measured birefringence of protein fibers in the thin lamellipodial region can be interpreted in terms of the number of filaments in the bundles. We confirmed that birefringent fibers are actin-based using conventional fluorescence-labeling methods. By recording movies of time-lapsed Pol-Scope images, we analyzed the creation and dynamic composition of radial fibers, filopodia, and intrapodia in advancing growth cones. The strictly quantitative information available in time-lapsed Pol-Scope images confirms previously deduced behavior and provides new insight into the architectural dynamics of filamentous actin.

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   Online version of this article contains video material. Online version available at www.molbiolcell.org.
^*   Corresponding author. E-mail address: rudolfo{at}mbl.edu.

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Molecular Biology of the Cell
Vol. 10, 197-210, January 1999
Copyright © 1999 by The American Society for Cell Biology

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