Charlotte Moore was born in Ercildoun, Pennsylvania. She graduated from Swarthmore College in 1920 and went on to Princeton to assist Henry Norris Russell. During this time, she worked at the Princeton University Observatory and the Mt. Wilson Observatory. She worked much on solar spectroscopy, studying the spectral lines of the Sun and identifying the chemical elements in the Sun. She earned a Ph.D. in astronomy in 1931 from the University of California, Berkeley on a Lick Fellowship. She then returned to Princeton.