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    On the Trope Namer 

(paraphrased) Hey, wouldn’t it be cool if the character turned out to be a woman?
Random staff membernote  during the development of the original Metroid. Now we're here.

Kid Icarus: Gosh-a-cus, Princess Lana! Samus is super-duper-a-cus!
Lana: Well... Samus is a veteran of many impossible missions! Samus is a super-powered cyborg! Samus is the greatest space hunter in the Galactic Federation! (Samus removes her helmet) Samus is a... woman?!
Kevin: Whew! You sure are!
Captain N: The Game Master comic story "Money Changes Everything"

Wasn't expecting me to be a woman, were you?
Samus Aran to Mario, Paper Mario X, Prologue: How to Steal a Castle

Whoa! Sorry about you suit... mam?! GUYS! We've been chased by a girl!
Sonic the Hedgehog, right after finding out Samus's gender, Paper Mario X, Chapter 61: The Mixed-Up Paths of Bowser's Castle

Pvt. Vasquez: Damn! (Beat) I totally mistook you for a man!
Samus: Samus is a woman's name.

Ange: No, how is that...
Tusk: It's a... It's a...
Momoka: IT'S A SHE!
Ange: You're a woman in that armor all along?!
Samus: Yes. Do you have a problem with that?
Metroid: Kamen Rider Generations Vol. 2 — Ex-Aid Era feat. Cross Ange, "Episode 002: Fallen Angel, part 2"

Kraid: I'm the strongest dragon that you've ever seen
You're gonna die, motherfucker, I take up five screens!
I'm gonna swallow you whole, and then you'll go down easy!
Then you'll be digested and converted to feces!
I'll take off your helmet, see the fear in your eyes
And your beautiful hair, and your well-toned thighs
And your pillowy lips and... wait, you're not a guy?
Samus: Is that a problem, you FUCK?!
Kraid: Uh, no, Prepare to Die!
Kraid was hesitating but Samus was set to fight
She got into a battle stance that made her butt look super-tight
Then Kraid said
Kraid: No wait, I think there has been a gigantic mistake
I actually just wanted to give you desserts I have baked!
But Samus said
Samus: What pisses me off most in this world
Is when enemies get nice when they all find out I'm a girl!
So do me a favor, and take your cakes and your pies,
And shove them so far up your ass that they end up behind your eyes!
Stop treating me like I'm a sex object,
Mother Brain's a woman and she gets respect!
Mother Brain: Yeah, Kraid, you never talk to me that way!
Kraid: Oh, gosh, I wonder if it's because you're a huge disgusting brain?
Starbomb, "Regretroid"

Luigi: It's not fair, Link. The only people who flirt with me are other guys.
Link: Listen, if this is about 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: Samus?
Luigi: That's it! I told him I didn't swing that way...
Link: (facepalming) 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!?

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?"

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.

Must restore peace! Must escape! Peace in the universe is once again restored! HOLYCRAPIMAGIRL!!!
Brentalfloss, "Metroid in 5 Seconds"

"I thought his name was Metroid."


    Anime and Manga 
Saionji: You're a girl?
Utena: I don't remember saying I was a boy!

[[Cagalli walks in wearing a fancy dress]]
Kira: You're... a girl.
Cagalli: YOU ALREADY KNEW THAT!
Kira: Well... yes, but... seeing you like this, just... reminded me.
Cagalli: Same thing, jackass!

Athrun: "You're a...a girl??"
Cagalli: "...Wh-what IS it with you men?!"

    Comic Books 
What? Everyone just assumed I was male. The ruthless, Nietzschean will to power, however, isn't gender-specific.
Deathmonger, Empowered vol. 9

    Comic Strips 
Kevin: You where a girl under that hat the whole time!
Kimmy: Kim is a ''girl's'' name
Kevin: I thought it was short for Kimothy!''

    Fanfiction 
Captain Borghese's wide-eyed stare finally tore away from staring at where the plasma blast had struck. He blinked in surprise to see the smiling woman revealed by the now transparent helmet.

Gloomtail: WHAAAAATTT? You destroyed my precious younger sister!
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...
Paper Mario X 2, Chapter 72: The Tail of Gloom and Its Doom

    Film — Live-Action 
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?

Neo: I just thought you were a guy.
Trinity: Most guys do.

Sgt. Hondo Harrelson: Sorry, wrong room.
Officer Sanchez: Who were you looking for?
Hondo: Chris Sanchez.
Sanchez: I'm Chris Sanchez.
Hondo: You're Chris Sanchez?

    Literature 
No wonder you go by Jack. I have never heard of anyone named Jacqueline who was not a giiirl oh crap.
Wreth, And Then There Were Monsters

The [power-armoured] figure halted a couple of metres away and, to my astonishment, addressed me by name.
"Hello Ciaphas," it said, though a vox unit on its chest. The voice sounded familiar, although I couldn't be entirely sure until a golden gauntlet rose to push back its visor. It opened, with a hiss of breaking atmospheric seals, and a well-remembered face, framed with golden hair, grinned at me, devilment dancing as always in the depthless blue eyes. "We really must stop meeting like this."
Clearly enjoying my stupedfied expression, which by this point I'm bound to admit would have done credit to Jurgen, Amberley's smile widened still further.
With all due modesty, I have to say I recovered remarkably fast under the circumstances.note 
Commissar Ciaphas Cain, Duty Calls

Kitai: Have I ever attempted to deceive you?
Tavi: The first night I met you, you gave me that [scar] with one of those stone knives. And I thought you were a boy.
Kitai: You are slow and stupid. We both know this. But have I ever deceived you?

    Myths & Religion 
And Bradamante, taking off her shield,
Had next removed her helmet from her head.
In doing so, the golden coif, which held
Her tresses coiled and flat, she likewise shed.
They fell about her shoulders and revealed
Her unmistakeably as a young maid,
Who was as beautiful in countenance
Orlando Furioso written in 1516

    Live-Action TV 
"You were so easily convinced that it had to be his son that was swinging the knife, but it's 2015, Emma. Sexist much?"
Piper Shaw, Scream: The TV Series
Myka Bering: Pete, Artie says that H.G Wells is actually....
Pete Lattimer: A woman, a really hot woman, maybe good with a gun.
Myka: He left out the gun part.

Myka Bering: H.G. Wells is actually a woman, I am going to have to process this.
Pete Lattimer: Make it fast would ya?

    Video Games 

Oh man, don't tell me I've been slaying hot girls this whole time.
Eddie Riggs, after one of the evil druids he was about to kill turned out to be an ally, BrĂ¼tal Legend

Hold on, the Assassin is a chick? How did I miss that?
The Assault, Monday Night Combat

Shadow Naoto: "Naoto"... Such a cool, manly name! But a name doesn't change the truth. It doesn't let you cross the barrier between the sexes. How could you become an ideal man when you were never male to begin with...?
Yosuke: Wait, what...? Did he... Did I hear that right!?
Kanji: H-He's not a guy!?

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...
Rival Schools: United By Fate

Max: You know, I would like to talk to you about various things so can you come to our ship, Sir Valbarossa?
Valbarossa: You'll abide by the proper etiquette if you're going to invite a lady.
Eshiria: Wha? You're a lady, Valbarossa?

Whoa! He's... a she!
Ratchet, on the Mysterious Thief/Angela Cross's true identity, Ratchet & Clank: Going Commando

Uh... You're a girl? Mkay, we were all under the impression that uh... Can you just give me a minute I need to call your parents, mkay? This will just take a second. [...] Your child has just let me know that, um, s-she's actually a girl. Oh, you knew that? Mkay. Well, no, no, of course it's- It's fine, it's just I-I believe everyone was under the impression that um... that he was a boy. I mean that she was a boy! Mkay, so the whole King and Stick of Truth thing was actually - Was actually a girl the entire time.
Mr. Mackey, if you make the New Kid a girl, South Park: The Fractured but Whole

Goofy: You're pretendin' to be a boy, aren't ya?
Sora: (Sora and Donald both gasp) You're a... girl!?
Mulan: You didn't notice?
Sora: (Sora and Donald shake their heads) Not me.
Mulan: I think it's working.
Mushu: I don't know - those two would fall for anything.

    Visual Novels 
Suzuha Amane: I'm the John Titor who's been posting on @channel.
Rintaro Okabe: ...I thought he was a guy.
Suzuha Amane: I fooled you? Guess my camouflage worked, then.

    Webcomics 

"No, Sir."
"That's 'No, Ma'am'"
"No, M... Wait, you're a woman?"

    Web Original 
Tucker: What is your problem?! Why do you even care if he's captured, I thought you hated that guy anyway for stealing your girlfriend?
Church: I never said I hated Tex, just that she was the reason we never got married.
Caboose: ..."She?"
[...]
Grif: Aha, I knew it! Only a chick could give me a headache this big!
Tex: What's the matter, never seen a girl before? How long have you guys been out here?

    Western Animation 
Batman: Red Claw is a woman?
Red Claw: Is that a problem?
Batman: Not at all. I'm an equal-opportunity crime-fighter.


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