He is known chiefly for his book Me'or Enayim (Hebrew, Light of the Eyes) in which he used critical methods to test the literal truth of the Aggadah, the non legalistic and narrative portions of the Talmud. His views were sharply criticised by Judah Loew ben Bezalel (the Maharal of Prague) in the latter's Be'er ha-Golah.
Dei Rossi's great work, Me'or Enayim ("Light of the Eyes") (Mantua, 1573-75; Berlin, 1794; Vienna, 1829; Vilna, 1863-66), includes the two works already mentioned and a third entitled Imre Binah. The latter is divided into four parts; the first part contains a survey of the Jews at the time of the Second Temple, narrates the origin of the Septuagint, points out the contradictions between some of the beliefs of the Talmudists and the proved results of scientific research, records the origin of the Jewish colonies in Alexandria and Cyrene, chronicles the wars of Bar Kokhba against the Romans, etc. Dei Rossi quotes from the writings of Philo, whose orthodoxy he questions. He criticizes him for having allegorized Biblical narratives of facts, and points out that the Alexandrian philosopher never gives the traditional interpretation of the Biblical text.
In the second part Dei Rossi criticizes a number of the assertions of the Talmudists (some of the criticisms were already extant and many of his criticisms were repeated by later commentators), and gives explanations of various aggadic passages which can not be taken literally (as, for instance, the aggadah which attributes the death of Titus to a gnat which entered his brain while he was returning to Rome). The third part is devoted to a study of Jewish chronology and translations from the writings of Philo, Josephus, and others, with commentaries. The fourth part deals with Jewish archeology, describing the shapes of the priestly garments and the glory of the Second Temple, and giving the history of Queen Helen and her two sons.
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