Our recent observation that microtubules (MTs) are required for completion of division (abscission) led us to analyze MT organization during cytokinesis. Although many studies of MTs in sea urchin eggs have been done, computer-aided analysis of optical sections described herein reveals a new MT assemblage, which we call furrow MTs. This assemblage comprises bundles of MTs that lie in the cleavage furrow. Furrow MTs become apparent when the furrow has progressed approximately one-third of the way through the egg and persist to abscission. Furrow MTs are 8–24-µm long and arc across the base of the cleavage furrow. Acetylated tubulin is localized primarily in the furrow suggesting a distinct MT population. Three-dimensional analysis of optical sections suggests that furrow MTs are spatially distinct from midbody and astral MTs.