In 1845, a church building was finally erected at the present location, Church and Baltimore Streets (the lot was purchased for $450 in 1844). Although the church was only partially completed, Saint Luke’s hosted the Diocese of Tennessee's Diocesan Convention in July of 1846.

The original hand-pumped organ was installed in 1852, as was the church bell, which still calls the congregation to worship and tolls the death of parishioners at funerals, one strike for each year of the deceased person's life. Pews and the chancel completed the building and Bishop Otey consecrated Saint Luke’s on May 14, 1853. 

The unique brass altar cross and the brass alms basin in use today were presented to the church in 1867 by Tennessee Bishop, The Right Rev. Thomas T. Quintard. (The cross is a copy of one in a Westminster Abbey chapel.) The cross and alms basin were a gift of the British Duchess of Teck, who met the bishop while he was in England raising funds for the University of the South at Sewanee, Tennessee. The duchess suggested he give them to “one of his parishes.” Bishop Quintard chose Saint Luke’s.

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