<p>Abstract</p> <p>Background</p> <p>Infection with <it>Plasmodium berghei </it>is a widely used model of murine malaria and a powerful tool for reverse genetic and pathogenesis studies. However, the efficacy of <it>in vitro </it>reinvasion of erythrocytes is generally low, limiting <it>in vitro </it>studies.</p> <p>Methods</p> <p><it>Plasmodium berghei </it>ANKA-infected blood obtained from a susceptible infected mouse was cultured in various conditions and <it>in vitro </it>parasitaemia was measured every day to evaluate the rate of reinvasion.</p> <p>Results</p> <p>High quality culture media were used and reinvasion rates were improved by vigorous orbital shaking of the flask and increasing density of the medium with gelatin.</p> <p>Discussion</p> <p>Using these settings, reinvasion of normal mouse erythrocytes by the parasite was obtained <it>in vitro </it>over two weeks with preservation of the infectivity <it>in vivo</it>.</p