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Anime and Manga
- Yu-Gi-Oh!: Bishounen Duke Devlin gets a twitchy eye when he's losing to Yami at a game of Dungeon Dice Monsters (Which he invented). Ouch.
- Kagami does this occasionally in Lucky Star, when she's trying to deal with Konata.
- Kaname gets this a lot in the high school parts of Full Metal Panic!!. Mostly due to Sousuke.
- The otherwise unflappable Xellos got an eye-twitch in Slayers TRY after Filia called him "raw garbage".
- Natasha Zabikov gets one on Mobile Fighter G Gundam when a boy calls her "Old Lady."
- Miyabi in Ai Yori Aoshi.
- Nenene in R.O.D the TV, multiple times in furious frustration with her new roommates.
- In Sonic X, Knuckles' eye has been seen twitching when he was extremely angry on a few occasions.
- Hiro-chan in Narutaru has this shortly before she snaps.
- Sasuke does this a few times in part one.
- In Code Geass when Viletta has been convinced by Shirley to pose in an anime-tight bikini, she gets a full on dose of this. With a little Foreshadowing:
- Viletta: You'll die a thousand deaths for this, Shirley.
- Miss Wormwood has one in Calvin and Hobbes: The Series.
- HERZ: Asuka's right eye often twitches when she is angry or jealous. Many characters get scared or worried when they see her eye twitching.
"Oh no," said Kensuke, looking really worried.
"What's wrong? All the readings are fine," said Kaori.
"Look." Kensuke pointed to the videofeed's enlarged view of Asuka's face. Maya and Kaori leaned forward to take a closer look. Asuka's right eye and brow were definitely starting to twitch.
"How much longer!? I've got to get out here! That ho is after my husband again!"
- Queen Of All Oni: Jade has one when, after getting curbstomped by Brother Light while trying to take the first tablet, he just gives it to her, saying she could have just asked from the beginning.
- Hak Foo is starting to develop a regular one, whenever someone gets a mask instead of him.
- The Elements of Harmony and the Savior of Worlds: Rarity has one when Megan tells her she actually was in attendance at the first Grand Galloping Gala.
- Twilight Sparkle has one too, when Star-Swirl the Bearded was referred to as a "nut".
- Sonic Generations: Friendship is Timeless: When Spike pushes Amy's Berserk Button and leaves Sweet Apple Acres, one of Amy's eyes twitch as she follows the baby dragon.
- Super Mario Bros. Z: Yoshi when surrounded by mooks they hadn't seen earlier.
Film - Animated
- Sir Ruber of the movie Quest for Camelot, one of the characters with a permanent tic, starts out with a slight Twitchy Eye that gets worse and worse throughout the film, until his whole face is practically quivering.
- Ed the hyena from The Lion King. He's the least sane from their pack and quite a Cloud Cuckoo Lander.
- Master Shifu of Kung Fu Panda seems to have developed one of these, particularly after Po gets selected as the Dragon Warrior—a trait which appears to afflict him much more in FanFiction.
- The Incredibles: after Kari has been putting up with Jack-Jack throughout the whole of the movie, she manages to freak out the new 'babysitter' with one of these.
- Russell being dragged across the Big Bad's window induces a slight eye twitch in the latter in Up.
- A brief example appears in The Princess and the Frog: "I was startin' to think that wishin' on stars was for babies and.. crazy people.."
Film - Live Action
- Done dramatically by Gandalf whenever the Ring speaks in The Movie.
- The film version of Watchmen does this with Rorschach, though sparingly.
- Quirrell has this in Harry Potter and the Philosopher's Stone due to his nerves and his generally anxious manner.
- Chief Inspector Charles La Rousse Dreyfus in the Pink Panther movies.
- The anchorwoman in the film version of V for Vendetta was said to blink involuntarily when she reported a story she knew was a lie.
- Several characters played by Kitano Takeshi have a twitchy eye, most memorably Nishi in Hana Bi, whose eye twitch announces an imminent outburst of sudden violence.
- In The Kid, while trying to make a sandwich for his younger self, Bruce Willis's left eye starts to noticeably twitch.
- In Amélie, Amelie's mother has developed an eye twitch as a sign of shot nerves.
- Seed of Chucky: Glen, who is the genderless offspring between the eponymous killer doll Chucky and his dollified girlfriend Tiffany, always has a notable twitch in his/her eye whenever he's anxious or goes into an Ax-Crazy episode. He also tends to wet his pants.
- Johnny English gets twitchy-eyed whenever Mozambique is mentioned.
- Variation: In the documentary Cocaine Cowboys: Reloaded the eyes of people in still photographs will suddenly, subtlety move while they are being described as paranoid, high, murderous, or all three at once.
- A man went into a job interview and his eye winked through the whole process. The interviewer said "Look, you are well qualified, but I'm afraid that facial tic will throw off clients." The man replied - "Funny you mention that, because all I have to do is take aspirin and the winking goes away - watch" and the man began dumping out his pockets looking for aspirin. In most of his pockets, however, were condoms. Finally, once a pile of condom packages of every sort piled up on the desk the man found two aspirin, took them, and the wink totally stopped. "Well, that worked," the interviewer admitted, but why do you have all the condoms? We don't want our clients to think you're a womanizer!" "It's quite simple sir," the guy said, "have you ever walked into a drugstore, winking like crazy, and asked for aspirin?"
- In the novel The Golden Goblet, the main character's villainous half-brother has a permanent tic in one eye that makes him look even more sinister.
-Vetinari: I just can't stand to see his poor face twitch all the time.
- Mr Teatime in Hogfather has an odd twitch whenever his name is mispronounced.
- In Jingo Lord Vetinari tells Vimes that the reason he doesn't like Captain Carrot in the room when Vimes is being debriefed is that Carrot tends to eye-twitch whenever Vimes tells a direct lie.
- Prince Andrei Bolkonsky developed facial tics in War and Peace, but that was probably due to a degenerative disease. The way it's written sounds like his personality (he was a curmudgeony sort) was getting the best of him.
- The First Law: Glokta's ruined eye (along with his face, sometimes) tends to begin twitching/blinking uncontrollably when he's nervous, lying, or any other time it's inconvenient to look like he's not utterly composed. He Lampshades it in his thoughts, calling it "treacherous jelly."
- In the novelization of Resident Evil 0, it's mentioned that Birkin's paranoia has caused him to develop a facial tic. Though, it's a slight subversion because it's actually on the corner of his mouth rather than his eye.
- In Harry Potter and the Chamber of Secrets, Lockhart's Valentine's Day antics really irritate McGonagall.
Live Action TV
- Star Trek: Deep Space Nine featured Jack, a genetically-engineered super-genius who was unstable and borderline paranoid, in two episodes. He was very twitchy.
- Data's Evil Twin Lore in Star Trek: The Next Generation had a facial tic when first discovered and activated.
- Captain Darling from the fourth season of Blackadder! Actor Tim McInnerny had a hard time getting rid of this tic after filming ended.
- On MacGyver, Jack Dalton's left eye always twitches when he lies. It's so well known at the Phoenix Foundation there's a notice in the ladies' room warning about it.
- In Pushing Daisies, Ned's eye twitches when he lies.
- Barney in How I Met Your Mother does this occasionally, most notably when he was trying (and failing) to hide his fear at being on the receiving end of another slap from Marshall on Slapsgiving.
- Peter on The Monkees sometimes displays this tic when he is nervous or startled.
Davy: "Are you nervous?"Peter: "Nervous? Don't be silly! Look at how slowly I'm twitching."
- On Just Shoot Me!, Dennis realizes Nina is trying to keep a secret because of her "I've got an itch to snitch" twitch.
- Buffy the Vampire Slayer: Xander briefly developed an eye twitch every time anyone mentioned Faith, after they'd had sex together.
- Boy Meets World: Topanga begins to develop a really nasty one during the preparations for her wedding to Cory, which, naturally, are full of wacky hijinks as Eric... "helps."
- The Big Bang Theory: Sheldon has this (among other tics) when things don't go his way (for instance people interrupting him when he wants to show off his knowledge, or not being able to answer a question to a quizz).
- On The Nanny, C.C. Babcock has it happen whenever she sees Fran Fine and Maxwell Sheffield getting along. It first occurs in season one's "Here Comes the Brood".
Fran: (to the Sheffield children) Now everybody say goodbye to Daddy...C.C.: "Daddy"? Did I miss the wedding? (her eye twitches)
- In Family Matters, Carl developed one from dealing with Steve Urkel for so many years. It went away after a therapy session… until he saw his bill.
- It Ain't Half Hot Mum: Sergeant Major Williams' eye tends to twitch when ever he is starting to lose his temper. Which is to say, always.
- Fred Bonaparte in Psychonauts has eye twitches... even after he's cured by Raz.
- Boyd seems to have a twitchy eye as well. It's most noticeable when Raz first meets him outside the asylum ruins.
- Happens frequently throughout the Ace Attorney series.
- Count Veger in Jak and Daxter gets one when he finds out the Precursors are ottsels.
- A twitchy eye is one of the first signs you get that there's something that's not right about Lisa in Silent Hill.
- Portal 2 gives us a robotic example in Wheatley the Personality Core. He gets damaged early in the game and, from that point until the showdown with the Disc One Final Boss, he has a short circuit that randomly causes him to spasm and give off sparks. Since his body is intended to resemble a robotic eyeball inside a frame, the effect is strongly evocative of a twitchy eye in a human. This actually works as Foreshadowing that Wheatley is not as sane as he appears to be.
- In Mario Strikers Charged, this may happen to Luigi if a ball (or more than one ball) gets in the net. Yes, Luigi.
- The OCD-suffering Hannelore of Questionable Content does this when Marten unthinkingly jokes about spreading garbage around her apartment.
- This is supposedly the "tell" in poker of the Patterner in Exterminatus Now. To clarify: the Patterner is an Expy of Tzeentch, Warhammer 40,000's god of Magnificent, manipulative bastardry. When he's shown preparing to deal, he has at least five decks of cards floating around him and held in all four hands. And he displays he's bluffing when his eye (which is not only all of his head, but floats around separate from his body) twitches.
- Drive: Nosh does this when he learns that he has to go to Tesskil, whose inhabitants, the Tesskans, lost a war with his own species, the Veetans.
- Frankie and Stein's male half of the titular duo has an eyetwitch fairly regularly, generally when he has a wonderful idea. It also appears to be genetic, as it has been seen in his cousin. It may be contagious, as it has also been spotted in Karl's friend Adam, and the female half of the titular duo, Frankie.
- Jack's had a long two years on the run from the revenge driven Parcel Mistress in Homestuck, so the sudden appearance of these kids who he's pretty sure he killed already doesn't exactly help his mood.
- Karkat gets one in the Alterniabound flash game, while demanding that Gamzee stop dancing.
- The main character of Doctor Horribles Sing Along Blog has this as a Character Tic. It becomes very pronounced when he's nervous, almost to the point of being a Hollywood Tourette's tic. In particular, it tends to occur when he's discussing his Arch-Enemy Captain Hammer and when his desire for his Love Interest, Penny conflicts with his desire to be a supervillain. In contrast with the typical use of the trope, the tic becomes noticeably less pronounced the stronger he grows in his Dr. Horrible persona.
- Any character in Invader Zim will get twitchy whenever they start feeling crazy, but particularly Gaz and the so-happy-it's-freaky experiment, Nick. For many of the other "normal" humans, it can be interpreted as showing how the stress of living in such a Crapsack World has them perpetually near the breaking point, such that they twitch whenever something pushes them just a little bit further.
- Lex Luthor in Justice League gets this in "Injustice For All" when the other members of the Injustice Gang stare quietly at him for letting Batman alive instead of killing him.
- The Grim Adventures of Billy & Mandy uses this a lot. Mostly with Billy's mom and her Broken Smile.
- Mr DeMartino from Daria
- Brock Samson of The Venture Bros. has a very twitchy eye that manifests when he enters one of his Unstoppable Rages. This happens especially when killing people who have threatened his Family...or run him over with a truck, while he's returning the favor.
- When he appears in Poker Night 2, this is one of his "tells" to indicate he has a bad hand.
- Tweek of South Park, in addition to his overall jitteriness, has a perpetually twitchy eye.
- In a Simpsons parody of Amadeus, Lisa, as Salieri, does this when the emperor declares all music other than Beethoven obsolete, making her schemes against Mozart all for nothing. She then hails a coach for the nearest asylum and rides off, cackling hysterically.
- If you pay careful attention, you can see Ned Flanders twitch an eye during "Hurricane Ned" after the rebuilt house falls down, just before he puts his glasses back on and completely loses his shit.
- Yet another Simpsons example involves Moe the Bartender. When he attempts to remake his bar into a family restaurant and himself into a more family-friendly image, the customers—as per usual for Springfield—quickly annoy him to the breaking point. What sets him off on a G-Rated Cluster F-Bomb barrage is a child complaining, during Moe's twitch of anger, that her soda was too cold.
- "New Kids in the Blecch" has L.T. Smash do it when commandeering a navel ship to blow up the MAD headquarters.
- An episode of Futurama ends with a shot of Fry doing this after being forced to destroy his robot Lucy-Liu girlfriend, and then watch Bender start making out with the real Ms. Liu. Or at least her head. It's complicated.
- Wile E. Coyote often gets a twitch during his Oh Crap! moments.
- Megatron of Transformers Animated no doubt appreciates Lugnut's unswerving loyalty, especially in comparison with Starscream's constant backstabbing, but once in a while gets a twitching optical sensor when Lugnut overdoes the fawning adulation. "Just...go." * twitch twitch*
- Earlier in the franchise, Galvatron starts to exhibit a serious twitch towards the end of the fight with Hot Rod in Transformers: The Movie. Justified, given that his day has gone quite south, he's had his psychic leash yanked for much of the movie, and he later has a reputation for being, well, just plain nuts.
- Megas XLR has Magnanimous get one after being sucked through a portable black hole. And he lampshades it.
- Guess how Kevin can tell that his friend Ben is lying? It even becomes a minor plot point in one episode.
- Happens often in The Fairly OddParents; one example was when Timmy's mom showed him that her old bikini still fit.
- On Jimmy Two-Shoes, Heloise does this after Cerbee and Beezy ruin her private picnic with Jimmy. She then is paralyzed with rage, breathing deeply and heavily in one spot for hours.
- Atomic Betty: Maximus IQ does this after Galactic Guardian HQ is destroyed, because Noah and Minimus accidentally do it before he can.
- The lunch lady in the Fanboy and Chum Chum episode Little Glop of Horrors does this when Fanboy insults her glop.
- Not to mention Lenny, who even has an anti-stress twitch cream.
- Miss Achmetha from Veggie Tales has this as a character trait, and comes up pretty much every time she's used.
- A subtle one in Duke and the Great Pie War done by Duke (played by Larry):
- Shifu of Kung Fu Panda does this when Oogway chooses Po as the Dragon Warrior. This only serves as an early warning signal for the horror that he unleashes on Po.
- Used on My Little Pony: Friendship Is Magic almost as often as Quivering Eyes, usually when somepony is undergoing Sanity Slippage.
- In "Swarm of the Century" Twilight Sparkle audibly snaps and develops a twitchy eye when she sees the Parasprites eating Ponyville because of a spell she cast. In "Feeling Pinkie Keen", she develops a Twitchy Eye and evolves into Rapidash when Pinkie tells her that the Hydra wasn't the doozy she predicted.
- In "Suited for Success", Rarity earns one of these when she learns the famous fashion designer Hoity Toity is going to be attending her fashion show, and seeing the (tacky and overdone) dresses she made for her friends. She does it again at the end, when Hoity Toity agrees to sell her fashions at his shop, and proceeds to place a big order.
- Twilight briefly does this while worrying about whether Princess Celestia will approve of her friends in "A Bird In The Hoof".
- Spike's eye also twitches in "Owl's Well That Ends Well". Friendship Is Magic gets a lot of this.
- In "Party of One", Pinkie Pie goes crazy when none of the other ponies want to come to her party, and holds a tea party with inanimate objects complete with Creepy Circus Music and eye twitches.
- At the Grand Galloping Gala in "The Best Night Ever", Fluttershy develops a Twitchy Eye, Evil Laughter, and Dramatic Thunder when none of Canterlot's creatures want to be her friends.
- in "", When the Mane 6 reveal that a tribe of buffalo has dragonnapped Spike and treenapped a tree Applejack was bringing to her cousin for his town's orchard, Applejack's cousin Braeburn's eye begins to visibly twitch in shock.
- Twilight does this repeatedly after going nuts in "Lesson Zero".
- Insane Rarity has this again in Sweetie Belle's Bad Future dream from "For Whom The Sweetie Belle Toils".
- In Inspiration Manifestation, as more of the tome's power takes hold, Rarity begins doing this.
- The Tick has the character of Big Shot, a No Celebrities Were Harmed version of The Punisher. He gets all eye-twitchy whenever he shoots, fights, or even says the word "gun". This is indicative of some pretty deep-seated mental trauma, as he is prone to collapsing into uncontrollable sobbing while screaming "Why didn't you love me, Mommy?!"
- Spongebob Squarepants does this when Patrick meets him in his house. "I'VE BEEN WAITING FOR YOU PATRICK!"
- The Powerpuff Girls: Buttercup does this in "Nuthin' Special" in a scene where her sisters keep matching her in showing off special powers.
- Roodaka's eye twitches in BIONICLE 3: Web of Shadows as she expresses her demand for the corpses of the Toa.
- In The Penguins of Madagascar Skipper gets this when trying (and failing) to stop a plumber from exposing them. That's not exactly what causes the twitchy eye, it's the fact that he thinks just like Skipper.
(repeated line)There's only one way to do the job, my way. The right way.
Does this eye look weird to anyone.
- Skipper even lampshades it.
- Captain Murphy of Sealab 2021 has this moment when his finds out his Happy Cake oven has gone missing.
- Mickey Mouse does this in the short "Fire Escape" when he sees the giant rabbit baby.
- Detective Piquel ("pih-kell" not "pickle"; basically an expy of Det. Valiant) develops one after about two days with his manic Toon partner Bonkers (averted with Bonkers' second, younger and more adaptable partner).
- Louise Belcher gets a twitchy eye on Bob's Burgers, especially when she's angry.