The genus Orchestina Simon is distributed worldwide and is characterized by having an enlarged fourth femur with which these species are capable of jumping. The genus is also characterized by having a well-sclerotized sperm duct, a near H-shaped arrangement of the eye group, a 4-4-3-3 pattern of raised receptors on the tarsal organs of the legs I–IV, respectively, and by lacking spines on all the legs. All these characters, together with molecular evidence, support the monophyly of the genus, as well as its placement as sister group of Oonopinae. Most American species of Orchestina inhabit the forest foliage and canopy, but in unforested areas they also occur in shrubs and grasses. In this work, we revise the American representatives of Orchestina in a comprehensive context for the first time. In the past, six species were known from the Americas: five from the United States and only one from South America, described from Venezuela. After the study of the principal collections of the world and several field trips to several South American countries, we describe 85 new species and redescribe all previously known species. Matching sexes was occasionally problematic; while females are very homogeneous in somatic traits, males may have modifications on different parts of the body, making the matching very difficult. Therefore, in this review 56 of the species are described from only one sex, whereas 20 unmatched species are informally described as morphospecies, pending the discovery of conspecific sexes. Two species, O. pavesiiformis Saaristo and O. dentifera Simon, originally known from Israel and Sri Lanka, respectively, are here reported as introduced in several countries in the Americas and other continents. O. justini Saaristo described from the Seychelles is here considered a synonym of O. dentifera. One species, O. truncata Wunderlich, previously known as a subfossil spider from Colombian Copal is here tentatively redescribed based on recent material from Costa Rica, Colombia, and Ecuador; the female is also described for the first time. The species list by country is as follows (numbers refer to records, independently of the locality of the type material): United States (9 species: O. utahana Chamberlin and Ivie, O. moaba Chamberlin and Ivie, O. obscura Chamberlin and Ivie, O. saltitans Banks, O. nadleri Chickering, the introduced O. pavesiiformis Saaristo, and three new species, O. quasimodo, O. kamehameha, and O. auburndalensis); Mexico (3 species: O. utahana Chamberlin and Ivie, and two new species, O. nahuatl and O. chaparrita); Guatemala (1 new species: O. guatemala); Costa Rica (3 new species: O. laselva, O. griswoldi, and O. chiriqui; and the previously known O. truncata Wunderlich); Panama (5 new species: O. chiriqui, O. labarquei, O. pan, O. campana, and O. galapagos); Jamaica (2 species, the introduced O. dentifera Simon and O. galapagos); Haiti and Dominican Republic (only the introduced O. dentifera Simon); Colombia (6 new species: O. filandia, O. zingara, O. arboleda, O. cali, O. platnicki, O. pakitza; and O. truncata Wunderlich, plus the morphospecies OMI020 and OMI038); Venezuela (7 species: O. saltabunda; and 6 new, O. venezuela, O. aragua, O. bolivar, O. maracay, O. ranchogrande, and O. neblina); Trinidad and Tobago (1 new species: O. kairi); Guyana (1 morphospecies: OMI026); Ecuador (18 new species: O. galapagos, O. fernandina, O. erwini, O. ecuatoriensis, O. sotoi, O. magna, O. shuar, O. golem, O. waorani, O. tzantza, O. predator, O. goblin, O. yanayacu,