- Adaptation Name Change: Only Murphy & Sparks retain their names from 2020 for the most part (Murphy's first and nickname were changed from Michael "Mike" to Hazel "Hank" and the name of Sparks' rank changed).
- Adaptation Personality Change: None of the personality traits from Sealab 2020 can apply to these characters.
- They Killed Kenny Again: Every episode or so, Sealab gets destroyed. Come the next episode, everything's back to normal.
- Unsympathetic Comedy Protagonist: Everyone except Quinn. And even he has his moments.
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Captain Hazel "Hank" Murphy
Voiced by: Harry GozSupposed to be the leader of Sealab, but is often acts like immature. When his voice actor died, he went to space to fight in the Great Spice Wars.
- Cloudcuckoolander: Just ask Bucket-head Wendy.
- Character Outlives Actor
- The Hero: (The original)
- Manchild: Oh yes.
- Psychopathic Manchild
Captain Bellerophon "Tornado" Shanks
Voiced by: Michael GozA retired football coach, came to Sealab as Captain Murphy's replacement after he went off to fight in the Great Spice Wars.
- Aerith and Bob: With a name like Bellerophon that a given. Though he explained that he and his several dead brothers are all named after characters in Greek mythology
- The Hero: (The replacement)
- Ooh, Me Accent's Slipping: Originally had a Southern accent, but later dropped it because it made people think he was gay.
- Sixth Ranger
- Suspiciously Similar Substitute: To Captain Murphy. Though he's more self-aware than Murphy.
Jodene Sparks (Commander)
Voiced by: Bill Lobley
- Affably Evil: Runs a supervillian operation on the side. But then again..
- Deadpan Snarker: Sparks invokes this usually when he's around Captain Murphy or many of the other crew members.
- Evil Cripple: Subverted. He's incredibly greedy, and may or may not run a supervillain operation. But, while he initially claims to be crippled, he later admits that he can walk just fine, he's just lazy.
- The Lancer
- Last-Name Basis: Really, if one didn't pause at just the right moment in "All That Jazz", they never would even know Sparks's first name.
- Manipulative Bastard: Oftentimes.Marco: Sparks, any idea what happened to [Hank's] happy-cake oven thing?Sparks: Uhh, yeah. I stole it and dumped it out in the ocean.Marco: What!? Why did you do that?!Sparks: Just part of a larger plan to drive him insane. Oh, I also have the orphans wet his bed while he's asleep.
- Only Sane Man: He's fairly smart but too lazy and greedy to be a responsible employee; but is Depending on the Writer on this.
Debbie DuPree (Lieutenant Commander)
Voiced by: Kate Miller
- Alliterative Name
- The Chick
- The Fundamentalist: In season 4 she adopts this trait for one episode, and it got brought up again briefly in later ones.
- My Biological Clock Is Ticking: "Chickmate" centered about hers going off, but at the end she decides she didn't need a baby because everyone else is a Manchild anyway.
- One Steve Limit: Shares a name with Debbie, but she is known as White Debbie.
Derek "Stormy" Waters (Lieutenant Commander)
Voiced by: Ellis Henican
- Back to School: Stormy never apparently got past fifth grade and is seen in class with fourth graders.
- The Big Guy
- The Ditz: He is pretty, but also pretty stupid.
- Pretty Fly for a White Guy: Tried to flash a gang sign to Black Debbie, but everyone else wasn't impressed.
Dr. Quentin Q. Quinn (Lieutenant Commander)
Voiced by: Brett Butler
- Alliterative Name
- Ignored Expert
- Last-Name Basis: Always referred to as Quinn. His first name's only seen on a computer monitor in "All That Jazz" and on his insurance policy in "The Policy".
- Omnidisciplinary Scientist: Has four Ph.Ds.
- Only Sane Man: Most level headed out of Sealab, but Depending on the Writer.
- Robotic Reveal Outed him as one, when he was trying to stop a conversation.
- The Smart Guy: Has an IQ of 260.
Marco Rodrigo Diaz de Vivar Gabriel Garcia Marquez (Lieutenant Commander)
Voiced by: Erik Estrada
Hesh Hepplewhite (Lieutenant)
Voiced by: MC Chris
- A Wild Rapper Appears!: Often Hesh's role in an episode.
- Alliterative Name
- Guest Star: Voiced by MC Chris.
- Sixth Ranger
- Streetwalker: Hinted to have been one in "Dearly Beloved Seed", as he fell 'off the wagon' and prostituted himself to a man. The original version of the episode made it clear he had been, as that was how he and Debbie met.
- Third-Person Person: HESH WANTS SOME SEX!
- Wholesome Crossdresser
Debbie Allison Love (Lieutenant)
Voiced by: Angela Gibbs
- Ascended Extra: She started getting more screentime about the time Murphy left the show.
- As Herself: In "Return To Oblivion" Debbie Love is apparently playing herself, as seen in the Chunk opening credits.
- One Steve Limit: Shares a name with Debbie, but is known as Black Debbie. This causes confusion by Stormy.
- Passionate Sports Girl: Was a star quarterback, and even won the Heisman Trophy, much to Shanks's delight.
Dr. Ilad Virjay (Unknown Rank)
Voiced by: Adam Reed
- Ascended Extra: Right about the time Murphy left the show he started showing up in more episodes.
- Funny Foreigner: Has an Indian accent and is the doctor of the lab.
- Hidden Depths: Leads a rockabilly band called the Mahatmamas
- Morally Ambiguous Doctorate: It's not even clear just what his degree is in, and there have been plenty of times where he violated the Hippocratic Oath out of apathy.
Dolphin Boy (Lieutenant Jr. Grade)
A boy that wanders around Sealab, but speaks in chirps.