Barney Miller frequently featured LGBTQ+ characters, portraying them with dignity and complexity. The show consistently framed prejudice and discrimination as external problems, using these characters' stories to critique societal intolerance and advocate for fair treatment, particularly through the empathetic lens of Captain Miller, resulting in a net positive impact.

Barney Miller featured a character in one episode who was a man dressing as a woman, which served as a plot device. The show handled the character with a degree of humanity, avoiding outright villainy or mockery. However, the portrayal focused more on the police officers' reactions and discomfort, rather than offering an affirming or deeply explored depiction of the character's identity, resulting in a neutral net impact.

The series portrays Jewish characters, most notably Detective Fish, with warmth and humor. While his Jewish identity is a consistent aspect of his character, any humor associated with it stems from his personal quirks and experiences rather than a critique or negative stereotype of the faith itself.