Stewie Griffin
Stewie's voice is one of the cornerstones of Family Guy itself. In addition, the character's development from a severely evil genius into a sarcastic mess is one of the best in animated television. It became clear early on in the show's run that Stewie's plans for world domination could only go so far while remaining funny, so MacFarlane and company tweaked the character to give him more depth. While Stewie's intelligence has remained startlingly high, his loner attitude has fallen by the wayside. As such, he's developed entertaining partnerships with various Family Guy cast members, none of which are more notable than his friendship with Brian.
Stewie Griffin smiling in Family Guy
Similar to his role as Peter Griffin, MacFarlane has to cover a wide range of emotions as Stewie. While some scenes may be in line with Family Guy's famous irreverence, other sequences call for a much more emotional approach. Without Stewie, the show would lose a huge number of its comical moments and its heart. As such, Seth MacFarlane's role as Stewie Griffin is his most valuable acting performance in Family Guy.
Stewie Griffin choking on a grape in Family Guy
Peter Griffin
MacFarlane perfectly encapsulates Peter's background by supplying the character with an overwhelmingly strong Bostonian accent. However, it's not just Peter's dialogue that makes Seth MacFarlane's performance as Peter Griffin so impressive. As well as performing many musical numbers as Peter, MacFarlane has also developed an iconic laugh for his main character.
Peter Griffin in Family Guy holding a baseball bat in a school corridor
Despite being set in the fictional New England town of Quahog, Peter is the only main Family Guy character with an accent that reflects his roots, but that only helps him stand out more. Peter also has a tendency to go from child-like wonder to existential misery at the drop of a hat, and it's Seth MacFarlane's acting range that allows him to do it all believably. In addition, some of Peter Griffin's best quotes in Family Guy are only so highly revered because of MacFarlane's acting.
Peter Griffin celebrating on Family Guy
Glenn Quagmire
Quagmire probably speaks the fastest out of all of Family Guy's characters, let alone the group given life by MacFarlane. Glenn Quagmire wears many hats throughout the show, including that of an airline pilot, Peter's drinking buddy, and a ladies' man. As such, he adopts a different persona depending on his current setting. Seth MacFarlane is a master of consistent performances when it comes to voice acting, but Quagmire is one of his ultimate triumphs when it comes to flexibility. The character also has a relatively short fuse, and his raging outbursts are some of the best sequences because of MacFarlane's delivery.
Glenn Quagmire looking annoyed as he hosts a trivia night in Family Guy
Jasper
Brian isn't the only talking dog in Family Guy, as his cousin Jasper is often forgotten. Although used sparingly, Jasper has appeared several times throughout the show. Despite being a relation of Brian, Seth MacFarlane does an admirable job of making the two characters sound nothing alike. Their overall demeanor and mannerisms couldn't be more disparate, but they still interact well whenever they share a scene.
Jasper gasping in Family Guy
Carter Pewterschmidt
In an arguably uncomfortable casting move, Seth MacFarlane plays Lois Griffin's father as well as her husband. However, the two characters couldn't be more different if they tried. Where Peter finds joy in almost everything he does, Carter revels in misery and seemingly likes very little. The character of Carter Pewterschmidt gives Seth MacFarlane the chance to play a deeply pessimistic character, which isn't a common occurrence for the actor. Luckily, the negativity of Carter works well when played against other Family Guy characters.
Carter Pewterschmidt yelling at Brian and Stewie Griffin in Family Guy
Seamus Levine
There is a counterpart in Family Guy for a great many characters from The Simpsons. The Sea Captain from The Simpsons and Seamus Levine from Family Guy have a great deal in common, including their voices. Both characters speak with the seasoned growl of a sailor, but Seamus' all-wooden limbs set the two men apart. MacFarlane likely based his performance as Seamus on Hank Azaria's Sea Captain, but the imitation is sufficiently different to make the two figures independent of one another.
Meg Griffin on a candle-lit date with Seamus Levine in Family Guy
Tom Tucker
Seth MacFarlane's news anchor cadence is very accurate, if not parodying the profession a little. Tom Tucker has been shown outside the newsroom on numerous occasions, and he's often more comfortable conversing with others as if he's still in front of the camera. MacFarlane keeps Tom Tucker's speech patterns very similar for comedic effect - as if the character's media training has taken him over completely. MacFarlane has made effective use of Tom Tucker's consistent communication style in various scenarios, including acting in Halloween movies and even in more intimate moments.
Tom Tucker with a beer looking angrily at Peter Griffin in Family Guy
God
The version of God in Family Guy is unusually jovial and uncharacteristically inappropriate at times. Seth MacFarlane's naturally deep voice lends itself well to voicing a character like God, but he injects his own twisted sense of humor into the show's iteration of the bearded entity. While Family Guy's God looks every inch the traditional representation, he's been portrayed as many things that don't align with how he's described in the Bible. God often interacts with other Seth MacFarlane characters, having regular disagreements with Peter Griffin and his friends.
God presenting a man in a glass case to a roomful of businessmen in Family Guy
Dr. Elmer Hartman
MacFarlane adopts a gritty, authoritative tone for his role as Dr. Hartman, which is actually at odds with the character's lack of professionalism. Dr. Hartman often asks his patients questions that he should really know the answer to, but he does so with an unnerving amount of confidence. The conviction in MacFarlane's voice as the character bumbles through medical examinations is a huge factor in what makes Hartman's scene so funny.
Lois laughing at Dr Hartman holding a clipboard in Family Guy