Afro Samurai does not feature any identifiable LGBTQ+ characters or themes. The narrative focuses exclusively on Afro's quest for revenge, with no explicit or implied queer representation within its storyline or character arcs.

Afro Samurai does not feature any identifiable transsexual characters or themes within its narrative. The story primarily focuses on Afro's quest for revenge and the pursuit of the Number One headband, without exploring aspects of gender identity.

The show features Lady Sio, a prominent female antagonist and skilled swordswoman. While she engages in direct physical combat with Afro Samurai, she is ultimately defeated by him. There are no clear instances of her or any other female character achieving victory over one or more male opponents in close-quarters physical combat.

The anime adaptation of Afro Samurai maintains the established genders of its core characters from the original manga. No characters canonically established as one gender in the source material are portrayed as a different gender in the show.

The characters in the Afro Samurai animated series, including the protagonist Afro, consistently maintain the racial depictions established in the original manga by Takashi Okazaki. No canonical characters were portrayed as a different race.