The collection of stained glass is one of the oldest and most important in Europe. Its origins go back to Grand Duke Ludewig I of Hesse, who presented the windows from the Church of the Knights' Chapter at Wimpfen (1270–80) to the museum in 1807.
Of particular interest are the hierartic head of a saint from Lorsch Abbey, the oldest preserved example of stained glass in Europe; a medallion showing the Rhenish half-length figure of an angel dating from circa 1250; a Cologne Calvary of the early 14th century; the stained glass windows from the parish church at Partenheim, Rhenish Hesse, from circa 1440–1450, and the Salzburg panes by the glass painter Peter Hemmel von Andlau from about 1480. Further examples of stained glass include the 16th-century cabinet panes painted in grisaille and the 17th-century guild signs from Strasbourg.
Stained glass and artistic glazing found their way into the bourgeois home over the course of the 19th century, where pictures were meant to conjure the ambience of domestic privacy, shielding from the outside world. The modern stained glass scene is represented with works by Georg Meistermann, Ludwig Schaffrath and Johannes Schreiter, among others.