AbstractActin has many diverse functions in the outer retina. To help elucidate its organization in this area, we have investigated the extent of its association with the actin cross‐linking protein alpha‐actinin. Ultrathin sections of chicken retina were double‐immunolabelled with monospecific antibodies against actin and alpha‐actinin. The highest relative amount of alpha‐actinin to actin label was measured in the adherens junctions between the individual retinal pigmented epithelial (RPE) cells and between the photoreceptor and Mueller cells; in the photoreceptor myoid; and in the RPE basal microvilli. The lowest amount was in the Mueller cell microvilli, the RPE apical processes, and in the photoreceptor ellipsoid. It is likely that the areas containing the highest ratio of alpha‐actinin to actin labelling are where the actin filaments are most highly cross‐linked into bundles and linked to the plasma membrane by alpha‐actinin. Actin filaments terminate in these areas, and, except for the myoid region, they are involved in cell‐cell or cell‐substrate