Ambrosius Bosschaert was a pioneer in the history of Dutch still lifes and a painter of joyful flower bouquets. He had an unerring awareness of composition, and delighted in combining flowers with a wide variety of colors and shapes to create a pleasing and uplifting visual experience. As in this exquisite work, Bosschaert generally arranged his blossoms symmetrically. Here, two spectacular blossoms, a yellow iris and a red-and-white striped tulip, surmount a bouquet composed of numerous species, among them roses, a blue-and-white columbine, fritillaria, grape hyacinth, lily of the valley, and a sprig of rosemary. A damselfly alighting on the iris and a butterfly on the cyclamen blossom that rests on the wooden tabletop further enliven the composition.

Following Antwerp’s reconquest by Spanish forces in 1585 and the subsequent expulsion of all non-Catholics, the Protestant Bosschaert family moved north to Middelburg in about 1589. Middelburg, a prosperous trading center and the capital of Zeeland, was renowned for its botanical gardens, the most important of which was established in the 1590s by the great botanist Matthias Lobelius. After 1602 this herb garden was transformed into a flower garden, and almost certainly filled with exotic species imported from the Balkan peninsula, the Near and Far East, and the New World. Bosschaert, who may have trained with his father, probably began his career making drawings of rare and exotic flowers in such gardens; he certainly used preparatory images drawn from life to compose his paintings, sometimes depicting individual flowers in reverse. His style of flower painting became more naturalistic over time, as he developed techniques for rendering petals with soft, velvety textures. He also introduced subtle tonal gradations in the backgrounds to enhance the sensation of light flooding the image.

Bouquet of Flowers in a Glass Vase occupies a special place in Bosschaert's oeuvre, not only because it was painted in the year of the artist’s death, but also because it contains a moving testimony to his enormous reputation. Inscribed on an illusionistic plaque attached to the table's front are the words: "C'est l'Angelicq[ue] main du gra[n]d Peinctre de Flore / AMBROISE, renommé jusqu'au Rivage Moré" (This is the angelic hand of the great painter of Flora, Ambrosius, renowned even to the banks of the Moré). The knowledge that the Moré River is located on the Gold Coast of Africa and that the name Ambrosius derives from the Greek word ambrotos, meaning "immortal," lends an even more laudatory note to the statement.