In Spain, during the 16th century, Francis Barnard (John Kerr) visits the castle of his brother-in-law Nicholas Medina (Vincent Price) to investigate the cause of the mysterious death of his sister, Elizabeth (Barbara Steele), who had been Nicholas Medina's wife. Both Nicholas and his younger sister, Catherine (Luana Anders), offer a vague explanation about Elizabeth having died from a rare blood disorder. However, when Nicholas responds evasively after Francis asks for specific details regarding the disease, Francis advises that he will not leave until he discovers the true reason his sister died.
During dinner with family physician Dr. Leon (Antony Carbone), Francis again asks about his sister's death. Dr. Leon tells him that his sister had died of massive heart failure, literally "dying of fright". Francis demands to be shown where Elizabeth died. Nicholas takes him to the castle's torture chamber. Nicholas reveals that Elizabeth, under the influence of the castle's "heavy atmosphere", became obsessed with the chamber's torture devices. After becoming progressively unbalanced, one day she locked herself into an iron maiden, and died after whispering the name "Sebastian". Francis refuses to believe Nicholas's story.
Francis tells Catherine that Nicholas appears to feel "definite guilt" regarding Elizabeth's death. In response, Catherine talks about Nicholas's traumatic childhood, revealing that their father was Sebastian Medina, a notorious member of the Spanish Inquisition. When Nicholas was a small child, he was playing in the castle's torture chamber when his father (also played by Price) entered the room with his mother, Isabella, and Sebastian's brother, Bartolome. Hiding in a corner, Nicholas watched in horror as his father repeatedly hit Bartolome with a red-hot poker, screaming "Adulterer!" at him. After murdering Bartolome, Sebastian began torturing his wife slowly to death in front of Nicholas' eyes.
After Catherine is finished telling Francis about Nicholas, Catherine and Francis are informed by Dr. Leon that Isabella was in fact not tortured to death, rather she was entombed behind a brick wall while still alive. Dr. Leon explains: "The very thought of premature interment is enough to send your brother into convulsions of horror." Nicholas believes that Elizabeth may have been interred prematurely. The doctor tells Nicholas that "if Elizabeth Medina walks these corridors, it is her spirit and not her living self."
Nicholas now believes his late wife's vengeful ghost is haunting the castle. Elizabeth's room is noisily ransacked and her portrait is found slashed to ribbons. Her beloved harpsichord plays in the middle of the night. One of Elizabeth's rings is found in the keyboard. Francis accuses Nicholas of planting the evidence of Elizabeth's "haunting" as some sort of elaborate hoax. Nicholas insists that his wife's tomb be opened. Inside the coffin, they discover Elizabeth's putrefied corpse frozen in a position of writhing horror, hands clawed and mouth wide open, as if in a final scream.
That night, Nicholas, now on the very edge of sanity, hears his wife calling him. He follows her ghostly voice down to the torture chamber. Suddenly, Elizabeth appears from out of the shadows, causing Nicholas to fall down a flight of stairs. She is alive, and she is met by her lover, Dr. Leon. Elizabeth, thinking Nicholas dead, taunts his apparent corpse about their scheme to drive him mad so the two lovers could inherit his fortune and estate. Nicholas opens his eyes and begins laughing while his wife and the doctor recoil in horror. Nicholas stands up and overpowers Dr. Leon, who attempts to escape but falls to his death. Nicholas then approaches Elizabeth, and promises he will torture her horribly.
Francis, having heard Elizabeth's screams, enters the dungeon to see what has happened. Nicholas is now gibbering with insanity and has become convinced he is his own father, the evil Sebastian Medina. He confuses Francis for Sebastian's brother, Bartolome, and knocks him unconscious. He straps him to a stone slab located directly beneath a huge razor-sharp pendulum and gags him with a red scarf. The cackling Nicholas slowly lowers the swinging blade closer and closer to Francis' torso. Catherine arrives just in time with Maximillian, one of the family servants. After a brief fight with Maximillian, Nicholas falls to his death, and Francis is removed from the torture device. As they leave the basement, Catherine vows to seal up the chamber forever. They slam and lock the door shut, unaware that Elizabeth is still alive, gagged and trapped in the iron maiden.
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