The painting depicts Ophelia singing while floating in a river just before she drowns. The scene is described in Act IV, Scene VII of Hamlet in a speech by Queen Gertrude. The episode depicted is not usually seen onstage, as in Shakespeare's text it exists only in Gertrude's description. Out of her mind with grief, Ophelia has been making garlands of wildflowers. She climbs into a willow tree overhanging a brook to dangle some from its branches, and a bough breaks beneath her. She lies in the water singing songs, as if unaware of her danger ("incapable of her own distress"). Her clothes, trapping air, have allowed her to temporarily stay afloat ("Her clothes spread wide, / And, mermaid-like, awhile they bore her up."). But eventually, "her garments, heavy with their drink, / Pull'd the poor wretch from her melodious lay" down "to muddy death".

Ophelia's death has been praised as one of the most poetically written death scenes in literature.

Ophelia's pose—her open arms and upwards gaze—also resembles traditional portrayals of saints or martyrs, but has also been interpreted as erotic.

There, on the pendent boughs her coronet weeds

Clambering to hang, an envious sliver broke;

When down her weedy trophies and herself

Fell in the weeping brook. Her clothes spread wide,

And, mermaid-like, awhile they bore her up;

Which time she chanted snatches of old tunes,

As one incapable of her own distress,

Or like a creature native and indued

Unto that element; but long it could not be

Till that her garments, heavy with their drink,

Pull’d the poor wretch from her melodious lay

To muddy death.