Each and every moment in System Shock 2 when Shodan would talk to you. Creepy shit. Also, being low on ammo and knowing that the door you're in front of holds some baddies in check, yet you must go through. That was the first game to ever make me not want to play, yet I kept going. That was an incredibly atmospheric game.
Runner up would be in Resident Evil 2, playing on a 5.1 surround sound system. Having completed most of the police station, thinking it is safe to walk around, and then the licker leaps through the one way mirror in the interrogation room. Holy fuck that scared the crap out of me. The controller flew from my hands and I proceeded to die gruesomely. Also the first time in the game you see a licker. It is crawling on the outside of the window and disappears from view.
Great games, those.
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