Using total internal reflection fluorescence (TIRF) microscopy to visualize cortical actin and microtubules in the Drosophila syncytial embryo

Abstract

The Drosophila syncytial embryo is a powerful developmental model system for studying dynamic coordinated cytoskeletal rearrangements. Confocal microscopy has begun to reveal more about the cytoskeletal changes that occur during embryogenesis. Total internal reflection fluorescence (TIRF) microscopy provides a promising new approach for the visualization of cortical events with heightened axial resolution. We have applied TIRF microscopy to the Drosophila embryo to visualize cortical microtubule and actin dynamics in the syncytial blastoderm. Here, we describe the details of this technique, and report qualitative assessments of cortical microtubules and actin in the Drosophila syncytial embryo. In addition, we identified a peak of cortical microtubules during anaphase of each nuclear cycle in the syncytial blastoderm, and using images generated by TIRF microscopy, we quantitatively analyzed microtubule dynamics during this time.