Representative of the Greek Revival style, the Chapel is a monumental temple form with a meetinghouse plan and modified Greek hexastyle portico. Decorative features include corner pilasters, fluted and tapered columns without an entasis, and a transom. The building's interior has a three-sided balcony, supported by slender cast-iron columns, and features a 17 x 23 1/2' painting of the interior of St. Peter's Cathedral. The original design included a square bell tower with freestanding columns, located above the vestibule.

The University's first chapel, constructed in 1808, was deemed too small by the 1820s. In 1832, James Carlton and Ross Crane constructed the extant chapel for $15,000. Although the Chapel was regarded as too small by 1860, attempts to convince the legislature to provide funds for a newer facility failed. During the Civil War, the Chapel became part of the Confederate hospital and temporarily housed 431 Yankee cavalry prisoners captured from Stoneman's Raiders until they could be transferred to the Andersonville prison. Originally used to signal class periods and chapel services, the bell and its tower were removed in 1913. In 1867, Daniel Pratt donated George Cooke's oil painting, which was severely damaged by fire and extensively restored in 1955 by Walter Frobos. Renovations efforts in the early 1970s and 1990s preserved the Chapel, which is currently used for lectures, concerts, and meetings.