The show is located inside of The Muppet Theater.
The audience is ushered into a reproduction of the Muppet Theater from The Muppet Show, complete with Statler and Waldorf (in audio animatronic form) in their box.The two curmudgons heckle and welcome the audience to the show. The Muppet Show logo is lowerd onto the stage and an audio animatronic version of Nigel the Conductor and the muppet orchestra rises from the floor. Kermit pops out of the "O" announcing "It's The Muppet Show Live at Muppet Studios! Yaaaay!!!". As the sign is raised up,the screen cuts to a video of The Muppet Show's opening theme from The Muppets with re-shot footage and a new comment from Statler and Waldorf. Kermit welcomes everyone to the show and introduces the first act. Sam the Eagle, Beaker, Link Hogthrob and Rowlf come out on stage and perform "Smells Like Teen Spirit". Mahna Mahna and the Snowths perform "Mahna Mahna" to the original audio track featuring Jim Henson's vocals. New footage of Mahna Mahna leaving the theater and stopping at a burger bar before calling the theater was shot. A video clip is shown from episode 119 of The Muppet Show where The Newsman reports that furniture is turning into monsters as a man watching at home is eaten by his living room. An audio animatronic Kermit sitting on a tree stump sings "Bein' Green". A video clip from the Muppets viral video, Ghost Hunters is shown. From inside the Muppet Mobile Lab, Bunsen and a penguin observe Beaker's trip into a haunted house. Scooter, Floyd and the Electric Mayhem come out onstage and perform "Mr.Bassman" to the original audio track featuring Richard Hunt and Jerry Nelson's vocals. A video clip from the Muppets viaral video, Popcorn is shown.The Swedish Chef prepares popcorn shrimp (or tries to) while performing the song "Popcorn" on various kitchen appliances. Miss Piggy comes out on stage and performs "Never Before, Never Again" accompanied by Rowlf on piano. Walter comes out on stage and performs The Whistling Caruso. Fozzie comes out onstage and performs a comedy act only to be heckled by Statler and Waldorf and is pulled off stage by a shepard's crook. A video clip from episode 201 of Muppets Tonight is shown. Dr. Phil van Neuter tells Mulch to put on a scary video, but Mulch accidentally shows a home movie from a Christmas party, where Dr. Van Neuter got drunk after drinking too much egg nog. It was only one hour and twenty-five minutes after the party began that Dr. Van Neuter passed out. Floyd, Animal, Zoot and Dr. Teeth come out onstage and sing "New York State of Mind". Kermit and Fozzie drive onstage in a replica of Fozzie's Studebaker and they sing "Movin' Right Along". A video clip is shown from episode 423 of The Muppet Show where what begins as a Pigs in Space sketch finishes with Veterinarian's Hospital. Kermit comes out onstage and sings "Caribbean Amphibian" with the help of Floyd, Janice, Animal and Zoot. A video clip from episode 204 of Muppets Tonight is shown. Clueless Morgan is going to knight school to learn jousting. Polly Lobster threatens to hit Clueless over the head with a frying pan if he doesn't sing. Clueless begins a heartfelt rendition of "Beautiful Dreamer", but Polly has lied, and hits him anyway. Mr. Callahan then hits Polly with another frying pan to punish him for lying.The Electric Mayhem then come out on stage and sing "Can You Picture That?". The Muppets then come out onstage and sing "Rainbow Connection".The curtain closes and the audience leaves while The Muppet Show's closing theme plays.
Right outside of the theater is a hall to the Muppet*Vision 3D and the Dr.Teeth and the Electric Mayhem Live theaters.
- The puppeteers who perform the Muppets in the live segments of the show are trained by the Jim Henson Company for authenticity.
- Whenever Rowlf, Floyd, Zoot, Janice, and Animal appear onstage, their audio animatronic versions are lowered into the floor.