A study was made of rouleaux formation between erythrocytes of different species when suspended in Ringer solution with polyvinylpyrrolidone added (PVP with M.W. 360 000, 4.0 g/l). PVP was shown previously to be a suitable asymmetric macromolecule for promoting rouleaux formation. For the present study fresh samples of blood were obtained from humans, cats, rats, mice, dogs, rabbits and guinea pigs. Initially homogeneous populations of cells were allowed to form rouleaux in the microscope chamber for the purpose of determining the average cellular dimensions for each species. Subsequently red cells from two species at a time were mixed in equal proportions to assess the degree of preference for the cells of one species to form rouleaux among themselves rather than with cells of the other species. The sequencing of the cells in the mixed rouleaux which formed on the coverslip was determined from photomicrographs, using the previously determined knowledge of the cellular dimensions. Although the visual impression was that the rouleaux were composed of a random selection of cells from the mixed population, a preference for cells to join in rouleaux with cells of the same species was demonstrated statistically in nine of the ten combinations tested. Only the rat–mouse mixture of cells was excepted, perhaps because these animals are closely related members of the same family.