Popeye made his film debut in Popeye the Sailor, a 1933 Betty Boop cartoon (Betty only makes a brief appearance, repeating her hula dance from Betty Boop's Bamboo Isle). It was for this short that Sammy Lerner's "I'm Popeye the Sailor Man" song was written. I Yam What I Yam became the first entry in the regular Popeye the Sailor series.

For the first few cartoons, the opening-credits music consisted of an instrumental of "The Sailor's Hornpipe," followed by a vocal variation on "Strike Up the Band (Here Comes a Sailor)," substituting the words "for Popeye the Sailor" in the latter phrase. After that the "I Yam What I Yam" tune was used as the theme song. As Betty Boop gradually declined in quality as a result of the Hays Code being enforced in 1934, Popeye became the studio's star character by 1936.

The character of Popeye was originally voiced by William "Billy" Costello, also known as "Red Pepper Sam." When Costello's behavior allegedly became a problem because of the MPAA Code, he was replaced by former in-betweener animator Jack Mercer, beginning with King of the Mardi Gras in 1935. Jack Mercer copied Costello's gravelly voice style familiar to audiences. Olive Oyl was voiced by a number of actresses, the most notable of which was Mae Questel, who also voiced Betty Boop. Questel eventually took over the part completely until 1938. William Pennell was the first to voice the Bluto character from 1933 to 1935's "The Hyp-Nut-Tist", after which Gus Wickie voiced Bluto until his death in 1938, his last work as the "Chief" in Big Chief Ugh-A-Mug-Ugh.

Thanks to the animated-short series, Popeye became even more of a sensation than he had been in comic strips. During the mid-1930s, polls taken by theater owners proved Popeye more popular than Mickey Mouse,^[10][11] and by 1938, polls showed that the sailor was Hollywood's most popular cartoon character, leaving Mickey in a third place (The second place was taken over by Donald Duck). Despite this, Popeye would lose that place beginning in 1942, when Bugs Bunny became more popular than Popeye was. In 1935, as Popeye was able to surpass Mickey Mouse in popularity, Paramount added to Popeye's popularity by sponsoring the "Popeye Club" as part of their Saturday matinée program, in competition with Mickey Mouse Clubs. Popeye cartoons, including a sing-along special entitled Let's Sing With Popeye, were a regular part of the weekly meetings. For a 10-cent membership fee, club members were given a Popeye kazoo, a membership card, the chance to become elected as the Club's "Popeye" or "Olive Oyl," and the opportunity to win other gifts.

The original 1932 agreement with the syndicate called for any films made within ten years and any elements of them, to be destroyed in 1942. This would have erased all Fleisher films, which are considered the best of the series. King was not sure what effect the cartoons would have on the strip; if the effect was very negative, King was very eager to erase any memory of the cartoons by destroying them. However, the films were not destroyed, either through oversight or because of their success.^{[citation needed]}

The Popeye series, like other cartoons produced by the Fleischers, was noted for its urban feel (the Fleischers operated in New York City, specifically in Broadway), its manageable variations on a simple theme (Popeye loses Olive to bully Bluto and must eat his spinach and defeat him), and the characters' "under-the-breath" mutterings. The voices for Fleischer cartoons produced during the early and mid-1930s were recorded after the animation was completed. The actors, Mercer in particular, would therefore improvise lines that were not on the storyboards or prepared for the lip-sync (generally word-play and clever puns). Even after the Fleischers began pre-recording dialog for lip-sync shortly after moving to Miami, Mercer and the other voice actors would record ad-libbed lines while watching a finished copy of the cartoon.^[12] Fleischer Studios produced 108 Popeye cartoons, 105 of them in black-and-white. The remaining three were two-reel (double-length)Technicolor adaptations of stories from the Arabian Nights billed as "Popeye Color Features": Popeye the Sailor Meets Sindbad the Sailor (1936), Popeye the Sailor Meets Ali Baba's Forty Thieves (1937), andAladdin and His Wonderful Lamp (1939).

The Fleischers moved their studio to Miami, Florida in September 1938 in order to weaken union control and take advantage of tax breaks. The Popeye series continued production, although a marked change was seen in the Florida-produced shorts: they were brighter and less detailed in their artwork, with attempts to bring the character animation closer to a Disney style. Mae Questel, who started a family, refused to move to Florida, and Margie Hines, the wife of Jack Mercer, voiced Olive Oyl through the end of 1943. Several voice actors, among them Pinto Colvig (better known as the voice of Disney's Goofy), succeeded Gus Wickie as the voice of Bluto between 1938 and 1943.

In 1941, with World War II becoming more of a source of concern in the United States, Popeye was enlisted into the U.S. Navy, as depicted in the 1941 short The Mighty Navy. His costume was changed from the black shirt and jeans he wore in the original comic to an official white Navy suit, which Popeye continued to wear in animated cartoons until the 1960s. Popeye periodically appeared in his original costume when at home on shore leave, as in the 1942 entry Pip-Eye, Pup-Eye, Poop-Eye, An' Peep-Eye, which introduced his four identical nephews, and in the 1950 and 1952 Famous cartoons Popeye Makes a Movie and Big Bad Sindbad, which featured clips from 1937's Popeye the Sailor Meets Ali Baba's Forty Thieves and 1936's Popeye the Sailor Meets Sindbad the Sailor respectively. (See: List of Popeye the Sailor theatrical cartoons (Fleischer Studios).)