| Actor | Character |
| Kyle MacLachlan, FBI agent | |
| Miguel Ferrer, FBI forensics specialist | |
| David Lynch, FBI regional bureau chief | |
| David Duchovny, cross-dressing FBI agent | |
| Michael Ontkean, Twin Peaks sheriff | |
| Harry Goaz, Twin Peaks deputy | |
| Michael Horse, Native American Twin Peaks deputy | |
| Kimmy Robertson, receptionist | |
| Sheryl Lee, murdered at start of series | |
| Ray Wise, father of Sheryl Lee's character | |
| Grace Zabriskie, mother of Sheryl Lee's character | |
| Sheryl Lee, cousin to other character played by Lee | |
| Warren Frost, town doctor | |
| Mary Jo Deschanel, wife of town doctor | |
| Lara Flynn Boyle, daughter of town doctor | |
| Richard Beymer, owns Great Northern Hotel | |
| David Patrick Kelly, brother of Beymer's character | |
| Jan D'Arcy, wife of Beymer's character | |
| Sherilyn Fenn, daughter of Beymer's character | |
| Robert Bauer, mentally handicapped son of Beymer's character | |
| Joan Chen, widowed sawmill owner | |
| Dan O'Herlihy, late owner of sawmill | |
| Piper Laurie, sister of sawmill owner | |
| Jack Nance, brother-in-law of sawmill owner | |
| Don S. Davis, Air Force officer | |
| Charlotte Stewart, wife of Air Force officer | |
| Dana Ashbrook, son of Air Force officer | |
| Everett McGill, owner of Big Ed's Gas Farm | |
| | Actor | Character |
| Wendy Robie, wife of Big Ed's Gas Farm owner | |
| James Marshall, nephew of Big Ed's Gas Farm owner | |
| Peggy Lipton, waitress at R&R | |
| Chris Mulkey, paroled husband of R&R waitress | |
| Heather Graham, younger sister of R&R waitress | |
| Eric Da Re, trucker, drug-runner, and abusive husband | |
| Madchen Amick, waitress at R&R, wife of trucker | |
| Walter Olkewicz, drug-runner, bartender at Roadhouse | |
| Michael Parks, dangerous brother of Roadhouse bartender | |
| Victoria Catlin, owner of One Eyed Jack's | |
| Phoebe Augustine, would-be victim of killer | |
| Russ Tamblyn, eccentric local psychiatrist | |
| Gary Hershberger, best friend of Air Force officer's son | |
| Catherine E. Coulson, carries around a piece of wood | |
| Lenny Von Dohlen, receives Meals on Wheels, horticulturist | |
| Ian Buchanan, lover of sheriff's receptionist | |
| Hank Worden, works in the Great Northern, no official name | |
| John Boylan, in charge of Twin Peaks | |
| Robyn Lively, lover of person in charge of Twin Peaks, seductress | |
| Annette McCarthy, rich woman, abused by husband | |
| David Warner, former business partner of late sawmill owner | |
| Kenneth Welsh, psychotic ex-partner of primary FBI agent | |
| Billy Zane, falls in love with Great Northern's owner's daughter | |
| Michael J. Anderson, speaks backwards in a red room | |
| Al Strobel, one-armed man OR the spirit that possesses him | |
| Frank Silva, killer spirit | |
| Carel Struycken, large supernatural being | |
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