He established his reputation as a composer with his publication of a book of madrigals in Venice in 1546, and in 1551 he took part in one of the most famous events in 16th century music theory, the debate between Vicente Lusitano and himself in Rome in 1551.
Only one keyboard instrument using his 31-note-to-the-octave system survives from the Renaissance: the ‘Clavemusicum Omnitonum Modulis Diatonicis Cromaticis et Enearmonicis', built by Vito Trasuntino of Venice in 1606 to play the diatonic, chromatic and enharmonic.
While Vicentino was known as a composer, and wrote two books of madrigals and motets in a harmonically sophisticated style, it was his work as a music theorist that gained him renown.
In the 1550s, in Italy, there was a surge of interest in chromatic composition, some of which was part of the movement known as musica reservata, and some of which was motivated by research into ancient Greek music, including modes and genera.
In L'antica musica ridotta alla moderna prattica, he mentioned that the strength of singing must respect carefully the text and passage being sung.
Vicentino's solution in effect divides the octave into 31 equal parts, with good intonation for the thirds and sixths but somewhat beating narrow fifths.