Born in Franklin County, Pennsylvania, Samuel Crothers was raised in Kentucky from 1787 until 1804. In 1804 he moved to New York to attend the Presbyterian theological seminary under the charge of Dr. John M. Mason. In 1809, following his seminary training, Crothers was licensed to preach. He soon accepted a call to serve as pastor of both the Associate Reformed Church of Chillicothe, Ohio, and the Hop Run Church just southeast of Greenfield. This post lasted until 1813, when he devoted his time fully to the Hop Run Church. In 1818, Crothers joined the Presbyterian Church, returning to Greenfield 1820 to organize the Greenfield Presbyterian Church.
