In experimental animals infected with Leishmania major, the size of cutaneous lesions of the parasite often does not correlate with the number of parasites within the lesion. Indeed, cutaneous lesions can heal, but still contain parasites. Thus, the ability to determine parasite burden in infected animals becomes important, especially when assessing vaccines that are intended to induce sterilizing immunity. Here, Hermenio Lima, Julie Bleyenberg and Richard Titus describe a simple technique for enumerating Leishmania in infected tissue. It is hoped that this technique will allow all researchers working with Leishmania (especially those in countries where leishmaniasis is endemic) to determine parasite burden easily in infected animals.