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Night Slashers is similar to Capcom's Final Fight series, which is an archetypal side scrolling beat'em-up game. One or two player characters move from left to right through each level (most of which are split into three or more scenes), fighting with the enemy characters who appear, until they reach a confrontation with a stronger boss character at the end of the level. Once that boss is beaten, the players automatically move on to the next stage. Enemies appear from both sides of the screen and from out of doorways or entrances set into the background, and the player(s) must defeat all of them to progress. If the players try to simply travel through the levels without fighting, the screen will stop scrolling until all current enemies have been defeated, before allowing the players to continue progress. Enemies may move outside the confines of the screen, but players may not. There is a time limit to each stage.
Unlike Final Fight, Night Slashers features three characters with larger move sets (including "smart bomb" or "screen zapper" type moves) and a horror theme. During the course of the game, players fight through hordes of attacking zombies and mutants, as well as other enemies such as werewolves and elementals. Bosses also include stereotypical monsters such as a mummy, a golem, and lookalikes of Count Dracula and Frankenstein's monster.
The player(s) play the role of one of three heroes. The game also has three different endings, one for each character:
- Jake (ジェーク) - an American monster hunter with cybernetic arms and mastery of the Lightning Element. His nickname is "The Psychic Cyborg".
- Christopher (クリストファー) - a European vampire hunter and martial artist who has mastery over Water and Ice.
- Hong Hua (趙.紅華) - an Asian female martial artist who is proficient with mystic arts and mastery of the Fire Element.
In the Japanese version, the blood and gore is uncensored (red blood instead of green, but in the overseas version, there is an option to change blood color and violence level). At the end of a melee attack, Christopher holds out a cross instead of a blue crystal ball. The "Go" arrow flips over to read "To Hell!" in blood. There are extra pictures and dialogue in the cutscenes.