James Fletcher Hamilton Henderson Jr. was born December 18, 1897, in Cuthbert, Georgia, to a school principal father and a piano-teacher mother who started both Fletcher and his younger brother Horace on the instrument by age six. He studied mathematics and chemistry rather than music, earning a bachelor's degree in the two subjects from Atlanta University in 1920. He moved to New York City that year intending to pursue graduate chemistry work at Columbia University, but found only a year of work as a laboratory assistant before taking a job as a song demonstrator for the Pace and Handy Music Company. When Harry Pace spun off Black Swan Records, the first record label owned and operated by African Americans, he made Henderson its musical director from 1921 to 1923, a job that had Henderson accompanying blues singers in the studio and touring with the Black Swan Troubadours behind Ethel Waters from October 1921 to July 1922. In April 1922, while touring through New Orleans, Henderson heard the young Louis Armstrong and tried to hire him; Armstrong turned him down unless Henderson would also take on drummer Zutty Singleton.

Henderson organized his own recording group in 1923, and in January 1924 it became the house band at the Club Alabam on West 44th Street, his first steady working band. A booking at the Roseland Ballroom that July led him to make Armstrong a second offer, and this time Armstrong accepted, joining the Roseland band on October 13, 1924, and helping turn it into what critics of the day called the best African American dance band in New York. Henderson's musical director and chief arranger through this period was saxophonist Don Redman, who reworked King Oliver's "Dippermouth Blues" into "Sugar Foot Stomp," recorded May 29, 1925, in New York with Armstrong, Joe Smith, and Elmer Chambers on trumpets, Charlie Green on trombone, Buster Bailey and Redman on reeds, Coleman Hawkins on tenor saxophone, and Kaiser Marshall on drums. Armstrong stayed only about a year, chafing at the band's written arrangements and its more genteel New York manners, and left in November 1925 to return to Chicago.

Through the second half of the 1920s Henderson's orchestra recorded prolifically, sometimes under his own name and sometimes under aliases such as the Dixie Stompers or Connie's Inn Orchestra to satisfy competing record labels; Discogs credits the Fletcher Henderson artist entry across more than 2,000 releases issued under upward of two dozen different billing names. Coleman Hawkins anchored the tenor chair from 1923 until 1934, the longest tenure of any Henderson sideman, while cornetist Rex Stewart, trumpeter Tommy Ladnier, trombonist Jimmy Harrison, and drummer Kaiser Marshall passed through the band across these years, and Redman's 1926 arrangement of "The Stampede" became one of the orchestra's most celebrated instrumentals. A 1928 car accident injured Henderson's shoulder, an event his wife Leora later blamed for a slackening of his business discipline, and after Redman left in 1927 Henderson took on more of the arranging himself, sharing the load from 1930 to 1931 with saxophonist Benny Carter and buying additional charts from freelance arrangers including John Nesbitt. The Depression hit the band hard: despite popularity at Harlem's Roseland and Savoy ballrooms, Henderson struggled with money and management and repeatedly lost sidemen to better-paying rivals.

Henderson's own arranging matured fully in the early 1930s, built on the sectional call-and-response he and Redman had pioneered, dividing the band into brass and reed choirs that traded riffs behind soloists such as Hawkins, Red Allen, Roy Eldridge, Chu Berry, and Ben Webster. "Down South Camp Meetin'" and "Wrappin' It Up," both recorded for Decca on September 11, 1934, and "Christopher Columbus," recorded March 27, 1936, in Chicago from a Chu Berry riff arranged by Horace Henderson, all rank among the finest documents of this mature style. By 1934, with his own band in financial trouble, Henderson began selling arrangements to clarinetist Benny Goodman at the urging of Goodman's brother-in-law and talent scout John Hammond, who needed fresh charts for Goodman's spot on NBC's "Let's Dance" broadcast. Henderson often transcribed the charts from his own band's recordings, sometimes with Horace's help meeting Goodman's demand for new material every week; Goodman's August 21, 1935, engagement at the Palomar Ballroom in Los Angeles, built substantially on Henderson arrangements including "King Porter Stomp," is commonly cited as the moment the Swing Era began in the popular imagination, and Goodman himself always credited Henderson for the material.

Henderson disbanded his own orchestra in 1939 and joined Goodman's organization directly as staff pianist and arranger, a role he held into the early 1940s even as he continued periodically reassembling bands of his own in New York, Chicago, and Los Angeles. During a mid-1940s engagement at Chicago's Club DeLisa, Henderson employed a young Sun Ra as an arranger. He toured with Ethel Waters again in 1948 and 1949, and his last working unit was a sextet that played New York's Cafe Society. A stroke in December 1950 paralyzed him partially and ended his performing career for good. Fletcher Henderson died in New York City on December 29, 1952, eleven days after his fifty-fifth birthday.

Working first alongside Don Redman and then largely on his own, Henderson built the basic architecture of big-band swing: a rhythm section of piano, bass, guitar, and drums supporting brass and reed sections that answered each other in call-and-response riffs behind soloists, rather than the collectively improvised front line of New Orleans jazz. Trumpeter Louis Metcalf recalled that "the sight of Fletcher Henderson's men playing behind music stands brought on a learning-to-read-music kick in Harlem which hadn't cared before it," crediting the band with spreading music literacy through the scene. Henderson's orchestra also functioned as a finishing school for major soloists, passing Louis Armstrong, Coleman Hawkins, Benny Carter, Roy Eldridge, and Chu Berry through its ranks over three decades. His arrangements, sold to and performed by Benny Goodman from 1934 onward, supplied much of the musical DNA of the swing era's commercial breakthrough, and historians rank Henderson alongside Duke Ellington as one of the two most consequential bandleader-arrangers of the pre-swing and swing periods, even though Henderson's own band never achieved comparable commercial success.