Mosquito insectaries must be maintained around the clock without interruption, due to the lack of preservation procedures. Long-term maintenance of mosquito colonies can lead to population bottlenecks, laboratory adaptation, loss of genetic
markers, etc., leading to colonies that are attenuated and different from their wildtype counterparts. Cryopreservation is an essential tool to combat these challenges that has been developed in other insects, including silkworms, fruit flies, and honey bees. Unfortunately, there is no robust method of cryopreservation for mosquitoes. This protocol details the short-term freezing of Anopheles stephensi (Mosquito) larvae; an initial step towards establishing a cryopreservation method for mosquito larvae.