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- Spider-Man: Aunt May fits parts of this trope, though she's a widow and went back to dating after Uncle Ben died. She was with Edwin Jarvis for a while, but then it turns out that that Jarvis ain't Jarvis. You're a Parker, May—your love life's gonna be chaos, it's the law! She's more of a Maiden Aunt in the newspaper comics than in the comic books.
- Anna Watson, MJ's aunt and May's best friend, is a Maiden Aunt to a certain extent as well (or at least the favorite relative and unmarried part).
- In Four Daughters, Aunt Etta Lemp lives with her brother Adam and looks after Adam's titular four daughters. Made even odder in that no explanation is given for the whereabouts of Adam's wife.
- American actress Zasu Pitts was typecast in later life as a Maiden Aunt, appearing in dozens of movies and television shows.
- Subverted in Aren't Men Beasts? (1937), in which the father of a slandered groom dresses up as a Maiden Aunt in order to clear his son of any wrongdoing.
- Aunt Minerva in The Gift of Love: A Christmas Story.
- When George Bailey gets the chance to find out how the world would have turned out if he'd never been born, he finds that his wife Mary had become a bitter, unhappy Maiden Aunt.
- Most 40s and 50s series comedies featured Maiden Aunts, including the Mexican Spitfire series (Aunt Delia).
- Tim Burton's Alice in Wonderland had Aunt Imogen. She wasn't so much "single" as "engaged to a prince who doesn't exist".
- The main character's aunt in Hausu.
- In The Heiress, Catherine ends up as a Maiden Aunt, doting on the children of her cousin Marian, who call her "Aunt Catherine".
- Aunt Fanny in The Magnificent Ambersons.
- Gene Kelly in Anchors Aweigh has Aunt Susie, who is not old but has a young nephew who she takes care of.
- Averted in A Brother's Price. Men are very rare, but the women cope with this by sharing a husband among sisters, meaning that if you have an aunt, it is more or less guaranteed that she is married, and considered one of your mothers. If she is called your aunt, she is the sister of your father, and likely married to the brother of your mothers, as families often swap brothers. There are unmarried women, but they are almost never aunts.
- Agatha Christie's Miss Marple plays this trope straight. When first introduced, she is a Victorian Aunt; in one of her last appearances in 1965, she alludes to having a Victorian Aunt. (Victoria reigned for 64 years until 1901, so if Miss Marple was born towards the end of the 19th century, as seems likely, her aunt(s) were almost certainly Victorian as well.)
- Dorothy L. Sayers' Miss Climpson. We don't meet any actual family members for her to be an aunt to, but in one book she pretends Chief Inspector Parker is her nephew.
- In Andre Norton's The Crystal Gryphon, Joisan's aunt, Dame Math, entered a religious order after an Arranged Marriage fell through upon the death of the groom. She left her order (but was still called 'Dame') after her brother was widowed, and ran his household for the rest of her life.
- The protagonist of Patricia Wentworth's Maud Silver mysteries: a governess who became a private investigator. She plays this trope straight — most of the stories show her writing letters to her nieces in her spare time, and she is at least an Edwardian (if not Victorian) throwback in terms of hairstyle, taste in interior decoration, and her love for Tennyson's poetry.
- Patricia C. Wrede's and Caroline Stevermer's Sorcery & Cecelia has Aunt Elizabeth (who is chaperoning Cecy back home in the country) and Aunt Charlotte (chaperoning Kate and Georgina in London). At one point one of the girls speculates that Aunt Elizabeth is still unmarried because of a Grave Disappointment in her Youth (which turns out to be true, in a manner of speaking). Subverted in that Elizabeth gets married to Mr. Wrexton in the end.
- Granny Weatherwax of Discworld plays this to the whole population of her home village of Bad Ass. She's by far the most suspicious and conservative of the witches seen, as well as one of the most powerful, and her title of "Granny" is definitely only honorary, considering that she can tame unicorns.
- L. M. Montgomery had a lot of these in her stories — but considering when they were written, that's not surprising.
- The Widow Douglas from Mark Twain's works set in St. Petersburg, MO should count: Elderly, Wealthy, Conservative, no family mentioned, cares for the main cast...
- Superficially following the schoolmarm stereotype is Harry Potter's Dolores Umbridge. When she's introduced, she's actually described as looking like "somebody's maiden aunt" and she affects the mannerisms of one, including an apparent love of cats. But it's only a thin veneer hiding a power-hungry sadist.
- Aunt Peace and Aunt Plenty in Louisa May Alcott's Eight Cousins and Rose in Bloom.
- The title character in Patrick Dennis' Auntie Mame is a notable subversion of the trope.
- Matilda in The Full Matilda, but without the caring part. If anything she's more of a Deadpan Snarker. The book deconstructs her reasons for being one.
- The Hardy Boys have Aunt Gertrude, a classic example of the trope. She often scolds the Hardys for their dangerous exploits while secretly cheering them on, and occasionally offers up some wisdom that gives them a hint of what direction to go next.
- Angus Wilson's short horror story Raspberry Jam involves a young boy going to visit two maiden aunts, but unknown to him they have recently been released from a mental asylum. It doesn't end well.
- In Emma, Miss Bates is a resident spinster who never married and there isn't any mention of romantic past. She's popular with people, takes care of her old widowed mother and adores her niece Jane Fairfax who is an orphan, but was adopted by her late father's friend.
- In The French Lieutenant's Woman, Aunt Tranter is an old maid, though she's referred to as Mrs Tranter, is a kindly woman and satisfied with her lot in life. She adores her niece Ernestina and she's a very good mistress to her servants. She's particularly fond of and even motherly to Mary (her servant girl).
- In Sue Grafton's alphabet series, Kinsey Millhone was raised by her Aunt Gin after the deaths of her parents.
- In the Colleen McCullough novel The Touch, the protagonist looks back over her 20-year loveless marriage and decides it was still better than being a maiden aunt in her native Scotland.
- In the Sweet Valley High novel The Wakefields of Sweet Valley, Ted Wakefield thinks he's being raised by one of these because his parents died in a train crash, but the reader knows that she actually is his mother and concocted the story to hide the fact that he's illegitimate.
- Adelheid von Stechlin is this to Woldemar in Der Stechlin.
- Gwen's great aunt Madeleine in The Ruby Red Trilogy who lives in the Multigenerational Household that's headed by her sister-in-law and Gwen's grandmother Lady Arista.
Live Action TV
- Miss Emily and Miss Mamie Baldwin on The Waltons.
- Aunt Bee on The Andy Griffith Show.
- Aunt Ellen on Brendon Chase.
- Aunt Harriet on the 1960s Batman live-action program.
- Nugie on The Gale Storm Show.
- Doña Clotilde in El Chavo del ocho is a surprisingly more sexual (or at least romantic) take on this, but her attempts of "seduction" (almost always involving hugging or bringing food) always fail with her intended target, and she is quite the traditionalist anyway.
- Invoked in an episode of 30 Rock. After one too many romantic failures, Liz gave up on dating and decided to start her "graceful transition into spinsterhood". This included buying a cat and naming it "Emily Dickinson" as well as joining a book club reading Murder on the Orient Express. Of course, she reverted to her normal self by the end of the episode.
- Hilda and Zelda of Sabrina the Teenage Witch come off this way, despite not being as old as this trope usually implies (or at least, not looking as old). Zelda was briefly married back in the Middle Ages while Hilda leaves the show after getting hitched in the later seasons.
- On Downton Abbey, Edith quotes this word for word after getting jilted at the altar.
- Charles Godfrey's sisters Cissy and Dolly are portrayed as this in Dad's Army. Although they have no nieces or nephews, they are nonetheless very nurturing, with Dolly's famous "upside-down cakes" mentioned in many episodes.
- Played for Laughs in an episode of The Worst Witch when the girls go on a camping trip. Miss Drill and the girls blend in as normal school people fine but Miss Hardbroom stands out like a sore thumb. So when they run into Canadian boyscouts, Miss Drill tells them that Miss Hardbroom is her Maiden Aunt who is with them for the good of her health.
- In "Baby It's Cold Outside," the woman goes through various relatives and their probable reactions to her spending the night at her boyfriend's house. She mentions that "my maiden aunt's mind is vicious."
- Aunt Abby and Aunt Martha Brewster in the play and movie Arsenic and Old Lace. Their special cordial is extremely special.
- Juliana Tesman in Hedda Gabler.
- The Nutcracker features two Maiden Aunt characters.
- Charleys Aunt gets a lot of mileage from this trope. Donna Lucia (although a widow, rather than never having married) is presumed by all five of the youngsters to be one, although that's because they've never met her. (She isn't, by a long shot.) Babbs plays her as one while masquerading as her. Mr Spettigue expected her to be one, so he never even considers that there might be an imposture going on. The real Donna Lucia takes full advantage of it.
- Alison's House: Aunt Agatha never married and never even left home. Her sole concern is protecting the memory of her sister Alison, a Captain Ersatz of Emily Dickinson who gained posthumous fame as a poet.
- Mrs. Crumplebottom from The Sims Specifically, Miss Crumplebottom in The Sims 1, who is the unmarried aunt of Mortimer Goth and sister to Cornelia Goth, though she is never seen interacting with her family
- Sialeeds in Suikoden V is this to the Prince. Very understandable considering what she, Arshtat, and Haswar had gone through. Come to think of it, Haswar does count for this trope as well.
- Wendy Oldbag (or Windy Old Bag?) from the Phoenix Wright series. She's often seen hitting on Miles Edgeworth, who's probably young enough to be her son.
- The Simpsons:
- Patty and Selma Bouvier were single at first and played aunts to the Simpson kids, though none of them were really fond of them. Selma ultimately had multiple marriages, none of which lasted very long. Patty apparently lived a life of self-imposed celibacy (despite dating Skinner for a while), but in late episode came out of the closet as a lesbian and nearly married before finding out her would-be bride was a man in drag.
- Great Aunt Gladys, Marge's aunt, seems to fill this trope. In her video will, she warns Patty and Selma about "not Dying Alone," and gives them her old grandfather clock.
- The Legend of Korra has Kya, who never married and seemingly never wanted to, and is much beloved by her brother's children; unlike most examples, she's a Cool Old Lady, described by Word of God as "kind of a hippie" and can kick butt if necessary.
- Given certain other revelations the creators made about her, there's a pretty good reason she never married and never wanted to.
- On Young Justice, Amanda Waller introduces herself to the Belle Reve prisoners by noting that "I am not your mother, your maiden aunt or your friend." When introducing the apparently more sympathetic Dr. Hugo Strange, she adds "he is your maiden aunt."