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Mizu is the main protagonist in the Netflix series, Blue Eye Samurai, who is voiced by Maya Erskine. The titular blue-eyed samurai, she is a half-Japanese, half-European sword-master who has sworn herself to a quest of vengeance.

Eiji the Swordmaker is a character in the Netflix series, Blue Eye Samurai, who is voiced by Cary-Hiroyuki Tagawa. A blind blacksmith, he is a master crafter of swords, and serves as a mentor and father figure to Mizu.

Ringo was born without hands, a disability that could have resulted in his being left to die as an infant (due to the stigma of bad luck in Japanese folklore). While his father spared him this treatment, he would beat him for asking too many questions, and constantly berated him in front of customers when apprenticing him at his soba tavern. It is unknown what happened to Ringo's mother, who is never shown, but she has presumably died by the series' start.

Madame Kaji is a supporting character in the Netflix series, Blue Eye Samurai, who is voiced by Ming-Na Wen. Madame Kaji is a woman who has managed to find autonomy over her life in a world hostile to women. Tart, unflappable, wise and unsentimental (can sometimes be charmingly persuasive). She worked her way up to this managerial role from the bottom up. As a brothel madam, she manages a group of prostitutes and as a Courtesan (called Oiran in Japan) she has more refined entertainment skills and training in the traditional arts than common prostitutes.

Hamata, also known as Boss Hamata, was a sadistic crime lord who controls most businesses in his port-town by extorting tribute payments. He commands a large gang of yakuza, known as the "Thousand-Claw Army", who murder anyone who deceives or defies him.

Abijah Fowler is a character in the Netflix series, Blue Eye Samurai, who is voiced by Kenneth Branagh. An Irish gun and opium smuggler and influential trader with a lust for violence, he leads a plot against the Shogunate with Heiiji Shindo and their co-conspirators, Lord Tokunobu Daiichi and the Shogun's advisor, Chiba. He is also possibly the biological father of Mizu. Fowler serves as the overall main antagonist of Season One.

Madame Kaji is a supporting character in the Netflix series, Blue Eye Samurai, who is voiced by Ming-Na Wen. Madame Kaji is a woman who has managed to find autonomy over her life in a world hostile to women. Tart, unflappable, wise and unsentimental (can sometimes be charmingly persuasive). She worked her way up to this managerial role from the bottom up. As a brothel madam, she manages a group of prostitutes and as a Courtesan (called Oiran in Japan) she has more refined entertainment skills and training in the traditional arts than common prostitutes.

Heiji Shindo is a black market middleman, trading prostitutes, opium, and rare goods from the West including illegal guns. One of his brothers (it is unclear if he is older or younger) runs the Shindo Dojo in Kyoto, and acts as an informant to him; the Shindo House, a brothel in Kyoto, is also owned by their clan. Heiji claims he saw enough 'truly great swordsmen' in his youth, to know his talents were elsewhere, so in spite of his samurai background he pursued a career as a merchant- a position that allowed him to amass great wealth, but that was considered the lowest in prestige of the four social castes.

Chiaki is a character in the Netflix series, Blue Eye Samurai, who is voiced by Mark Dacascos. Chiaki was an assassin who tricked Eiji into making him a sword; he said his father was a bookbinder who'd been slain by a drunk ronin, and Chiaki wished to avenge him, claiming he would at least have a chance with one of Master Eiji's blades, even if it cost him his own life. He later became the leader of the Four Fangs, a small group of mercenaries for hire who are feared through much of Japan.