Height: 169 cm

Sarutobi Ayame is a ninja from the Oniwabanshuu by training, but is currently working as an assassin. She also has a part-time job at the Kunoichi Café.

Since her initial appearance, she has developed a serious obsessive infatuation with Sakata Gintoki, to the point of even stalking him. She is severely myopic, to the point of being almost blind (cannot tell living things from inanimate objects) without her eyeglasses. It was also hinted that she doesn't see well even with her eyeglasses on, as she once mistook Kagura (dressed up as Gintoki) for Gintoki himself. However, this could solely be due to her infatuation with him. She has frequently clashed with Otae for various reasons.

Ayame has a playful attitude but gets serious when it comes to her work. She also seems to enjoy bondage and S&M. Although Gintoki is indifferent to her affections and even treats her badly most times (e.g throwing things at her whenever he senses her stalker presence), her 'M' or masochistic personality only makes her enjoy the punishment, and his 'cruelty' only serves to encourage her.