We developed a simple, fast, economical, and versatile reactivation method for the study of herpes simplex virus type 1 (HSV-1) latency in the rabbit ocular model. Intrastromal injection of sterile, deionized water induced reactivation and ocular shedding of latent HSV-1 in 21 of 27 eyes (78%) in 93% of New Zealand rabbits. Other control groups (eg, anterior chamber injection of sterile, deionized water; topical administration of sterile, deionized water; intrastromal injection of air; and intrastromal needle track), were less efficient in reactivating latent HSV-1. Although the mechanism of reactivation in this model is unknown, the reactivation signal may be related to the relative amount of corneal trauma.