Although bats are one of the largest groups within the class Mammalia and may carry several zoonotic diseases, basic information about their hematology is limited. In this study, hematocrit (Hct), total white blood cell counts (TWBC; leukocytes), and differential white blood cell counts (DWBC) of free-ranging Neotropical bats were quantified. Blood samples from 255 bats representing 26 species from the families of Emballonuridae (3 species; 33 individuals), Molossidae (2 species; 26 individuals), Mormoopidae (1 species; 1 individual), Phyllostomidae (18 species; 180 individuals), and Vespertilionidae (2 species; 15 individuals) were collected in a Caribbean lowland rainforest of Costa Rica. Hct was measured after centrifugation of microhematocrit capillaries, TWBCs were performed using the Unopette system and a hemocytometer, and DWBCs were performed on eosin methylene blue stained blood films. Hct of bats ranged between 51.8 ± 0.7% for Phyllostomus discolor (n = 27) and 65.8 ± 2.2% for Molossus sinaloae (n = 6). Bat species of the same taxonomic family had comparable TWBCs; these were lower for insectivorous emballonurid, molossid, and vespertilionid bat species than for mostly phytophagous phyllostomid bat species. However, Ectophylla alba (Phyllostomidae) exhibited exceptionally low TWBCs (836 ± 166 /μl; n = 10); this was less than half of the TWBCs of all other bat species, which ranged from 1,714 ± 297/μl for Molossus bondae (n = 20) to 7,339 ± 1,503/μl for Trachops cirrhosus (n = 6). Species with higher TWBCs tended to have lower Hct values. Overall, blood cell morphology was similar to other mammalian species. A large number of polychromatophilic erythrocytes and differences in lymphocyte morphology were noted. This study provides important hematological values for Neotropical bat species and significantly expands the knowledge on basal physiological measurements of Chiroptera.