Fluorescent powders are one of the most common external markers used to study mosquito behavior and ecology. For their reliable and practical use, it is important to evaluate their effect on biological parameters such as survival, blood-feeding, and mobility. We evaluated the effect of five different fluorescent powders (Day-Glo ® ECO Series) on the survival of different age cohorts, blood-feeding success, and tethered flight speed on Aedes aegypti (Linnaeus) adult females. For survival analysis, three cohorts (2-5, 6-9 and 10-13 days old) were marked and mortality recorded until all died. To examine the effect of fluorescent powders on female response to blood-feeding, the proportions of unfed, partially fed, and fully engorged females, after being exposed to host blood under two different time sets (20 and 40 min.), were compared. Their impact on female tethered flight speed was evaluated recording their flight for 30 min. with a flight mill. Survival distributions between treatments were not significantly different within each cohort. Blood-feeding was not significantly different among marked or unmarked females at both times of blood exposure, with the exception of Signal Green-ECO 18 and Ultra Violet- ECO 20 (at 20 and 40 min.), in which a higher proportion of partially fed females was observed compared to control females. In relation to flight performance, no statistically significant difference in mean tethered flight speed (m/s), among marked and unmarked mosquito groups, was observed. Our results indicate that the tested powders and application method have few significant impacts on Ae. aegypti survival, blood-feeding success and flight performance, and are thus suitable for investigations of mosquito biology in the environment.