He was the son of the painter and sculptor Pavlos Prosalendis the Elder and the father of the painters Aimilios and Pavlos Prosalendis the Younger. He took his first painting lessons at the art school which had been founded and directed by his father on Corfu and completed his lessons at the Fine Arts Academy of Venice, graduating with first prize. After quite a long period of residence in Venice, he returned to Corfu, while in 1865 he was appointed professor of painting at the School of Arts and settled in Athens. After teaching for a brief period he resigned and returned to Venice where he received a prize at the city’s International Exhibition. In 1870, at the urging of King George I, he settled permanently in Athens and undertook the decoration of the chapel of the Old Palace while in 1876, with the formation of the second chair of painting at the School of Arts, he was again appointed professor and remained in this position till his death.
Mainly a portrait painter, he was also involved with genre painting, but in a more limited way. He did the portraits of various eminent figures, as well as the veterans of the Greek War of Independence, combining an academic style with realistic rendering, while in his genre scenes the influences of his Italian training are obvious.