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"Oh, my giddy aunt" was a colloquialism that was often used as an exclamation of surprise.
By the Doctor
The Second Doctor used the phrase often. He said it to himself when he saw the side-effects of RTC units, (PROSE: Invasion of the Cat-People) to himself when he was surprised by a Kroton, (TV: The Krotons) to Sergeant Benton as a Gell guard tried to enter the TARDIS, (TV: The Three Doctors) to himself after escaping from a group of snake-like aliens (COMIC: The Forgotten) and to Dastari during an argument in a cellar. (TV: The Two Doctors)
A clone of the Second Doctor said it as Lucius threatened to devour him. (AUDIO: Survivors in Space)
The Eleventh Doctor said it while facing a Diceman with Henry Gordon Jago, (AUDIO: The Jago & Litefoot Revival) and when he'd simultaneously dodged the blasts of a gunstick, a cyber wrist blaster and a translation sphere. (COMIC: Doctor Who and the Last Stand)
The Seventh Doctor, when asked about his family, once listed only his granddaughter and a giddy aunt, "but she was probably a metaphor". (AUDIO: Forever Fallen)
By others
In the 1890s, Henry Gordon Jago said it when he saw one of the Wet Men. (AUDIO: Dead Men's Tales)
The Brigadier said this when he found out that Professor Rubeish was missing. (TV: The Time Warrior)
Mrs. Wibbsey exclaimed it to Buddy Hudson and a cultist when realising something was wrong with the Fourth Doctor. (AUDIO: Starfall)
Missy said it to Osgood while they were on a plane. (TV: Death in Heaven)