->"No, Sir."
->"That's 'No, Ma'am'"
->"No, M... Wait, you're a woman?"
-->-- ''Webcomic/LegostarGalactica''

->''My first great triumph against impossible odds brought me a lot of attention. I think my favorite part of the whole affair was revealing to the waiting dignitaries who I am under my suit... The phrase of the day was '''"He's a whaaa?"'''''
-->-- '''[[TropeNamer Samus Aran]]''', ''{{VideoGame/Metroid}}: [[http://bobandgeorge.com/comics/metroid Third Derivative]]''

->''Must restore peace! [[CollapsingLair Must escape]]! Peace in the universe is once again restored! HOLYCRAPIMAGIRL!!!''
-->-- '''Music/{{Brentalfloss}}''', ''{{VideoGame/Metroid}} in 10 Seconds''

->''Whoa! Sorry about you suit…''mam''?! GUYS! We've been chased by a girl!''
-->-- '''Franchise/SonicTheHedgehog''', right after finding out [[TropeNamer Samus]]'s gender, ''FanFic/PaperMarioX'', Chapter 61: The Mixed-Up Paths of Bowser's Castle

->''Wasn't expecting me to be a woman, were you?''
-->-- '''[[TropeNamer Samus Aran]]''' to [[Franchise/SuperMarioBros Mario]], ''FanFic/PaperMarioX'', Prologue: How to Steal a Castle

->'''Luigi''': It's not fair, Link. The only people who flirt with me are other guys.\\
'''Link''': Listen, if this is about [[NoodleIncident that incident in '93]], it was a really awkward time for me and I didn't --\\
'''Luigi''': No. This ''bounty hunter'' guy tried to pick me up at the bar last night. Said he was only ''planetside'' for the evening and wanted to show me the time of my life. Couldn't even see his face under all that ''orange armor''...\\
'''Link''': Wait... Did this "guy" give you his name?\\
'''Luigi''': Yeah. Began with a 'S'. Sam... or something.\\
'''Link''': ''[[TropeNamer Samus]]?''\\
'''Luigi''': ''That's it!'' I told him I didn't swing that way...\\
'''Link''': ''({{facepalm}}ing)'' Oh, by Ganon's ''pork fetish'', Luigi...\\
'''Luigi''': What? What is it?\\
'''Link''': ''Nothing''. Not a ''gosh-darn thing''. ''Say!'' You wanna come over and cry yourself to sleep on my couch for ''no reason whatsoever!?''
-->-- ''Webcomic/ItSucksToBeWeegie'', "[[http://www.interrobangstudios.com/potluck/index.php?strip_id=1084 Flirting with Danger]]"

->''But no living man am I! You look upon a woman. Éowyn I am, Éomund's daughter. You stand between me and my lord and kin. Begone, if you be not deathless! For living or dark undead, I will smite you, if you touch him.''
-->-- '''Éowyn''', ''[[Literature/TheLordoftheRings The Lord of the Rings]]''

->'''Akira:''' Brother, you're alive!\\
'''Daigo:''' I'm sorry, Akira. It must have been tough for a lady like yourself...\\
'''Edge:''' What? His voice... it sounds like...!\\
'''Gan:''' A lady like yourself?!\\
'''Akira:''' I'm sorry I've been deceiving you. I'm not the younger brother of the big boss... I'm his younger sister...
-->-- ''VideoGame/RivalSchools: United By Fate''

->''Oh no, please don't tell me I've been slaying hot chicks this entire time.''
-->-- '''Eddie Riggs''', after one of the evil druids he was about to kill turned out to be an ally, ''VideoGame/BrutalLegend''

->'''Neo:''' I just thought you were a guy.\\
'''Trinity:''' [[ThereAreNoGirlsOnTheInternet Most guys do.]]
-->-- ''Film/TheMatrix''

->''Hold on, the Assassin is a chick? How did I miss ''that''?''
-->-- '''The Assault''', ''MondayNightCombat''

->'''Tony Stark:''' Good God, you're a woman! I honestly- I couldn't have called that. I mean, I would apologize, but isn't that what we're going for here? I thought of you as a soldier first.
->'''Ramirez:''' I'm an airman.
->'''Tony Stark:''' Well, you have actually excellent bone structure there. I'm kinda having a hard time not looking at you now. Is that weird?
-->-- ''Film/IronMan''

->'''Gloomtail''': WHAAAAATTT? You destroyed my precious younger sister!\\
'''[[Franchise/TheLegendOfZelda Link]]''': Hooktail WAS a girl!\\
'''Flurrie''': Wait, you're Hooktail's brother?\\
'''{{Kirby}}''': But what if "he" is a girl too…\\
'''Flurrie''': No, I'm sure this guy's a…guy…
-->-- ''FanFic/PaperMarioX 2'', Chapter 72: The Tail of Gloom and Its Doom

->'''[[TheHero Chrom]]''': Wait, you're...you're a woman?\\
'''[[spoiler: "Marth"]]''': [[LampshadeHanging And quite the actress, too.]] [[PaperThinDisguise Honestly, I'm surprised you didn't figure it out until just now.]]
-->-- ''VideoGame/FireEmblemAwakening''

->Chuck: Are you a guy, or a lady? Because I never heard the name Samus before so I don't know if it's a male or female name, and your voice filter sounds kinda unisex so I have no way to tell.\\
(Samus Stares for a while)\\
Samus: Female. If that even matters.\\
Chuck: Thanks, sorry. Just wanna make sure I don't misgender anyone by accident.

-> '''Sgt. Hondo Harrelson:''' Sorry, wrong room.\\
'''Officer Chris Sanchez:''' Who were you looking for?\\
'''Hondo:''' Chris Sanchez.\\
'''Sanchez:''' I'm Chris Sanchez.\\
'''Hondo:''' ''You're'' Chris Sanchez?
--> -- ''Film/{{SWAT}}''