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An iconostasis is a wall in an Orthodox church that consists of icons and other structural elements generally having a wooden frame and magnificently adorned with metal decorations. It symbolically separates the nave, or the human world, from the altar, or the heavenly world. The icons in an iconostasis are views into that heavenly reality. They are usually depictions of Biblical scenes, such as The Holy Communion and holy figures, such as Mother of God, Christ Pantocrator or the four Evangelists. In the centre of the iconostasis are Holy Doors, also known as Beautiful Gates or Royal Doors.

During the period of autonomy, there was a small church in the Imperial Palace where the library is today. There was an iconostasis there too, although it had been painted on canvas in imitation of a real structure, and was in fact a travelling iconostasis. Executed in the Italianate academic Petersburgian style, it was probably completed in the early 19th century. The iconostasis was acquired for the Imperial Palace in 1834, from whence it was moved to the collections of the National Museum in 1919.