Emilio Gino Segrè was born into a Sephardic Jewish family in Tivoli, near Rome, on 1 February 1905, the son of Giuseppe Segrè, a businessman who owned a paper mill, and Amelia Susanna Treves. He had two older brothers, Angelo and Marco. His uncle, Gino Segrè, was a law professor. He was educated at the ginnasio in Tivoli and, after the family moved to Rome in 1917, the ginnasio and liceo in Rome. He graduated in July 1922 and enrolled in the University of Rome La Sapienza as an engineering student.
In 1927, Segrè met Franco Rasetti, who introduced him to Enrico Fermi. The two young physics professors were looking for talented students. They attended the Volta Conference at Como in September 1927, where Segrè heard lectures from notable physicists including Niels Bohr, Werner Heisenberg, Robert Millikan, Wolfgang Pauli, Max Planck and Ernest Rutherford. Segrè then joined Fermi and Rasetti at their laboratory in Rome. With the help of the director of the Institute of Physics, Orso Mario Corbino, Segrè was able to transfer to physics, and, studying under Fermi, earned his laurea degree in July 1928, with a thesis on "Anomalous Dispersion and Magnetic Rotation".
After a stint in the Italian Army from 1928 to 1929, during which he was a commissioned as a second lieutenant in the antiaircraft artillery, Segrè returned to the laboratory on Via Panisperna. He published his first article, which summarised his thesis, "On anomalous dispersion in mercury and in lithium", jointly with Edoardo Amaldi in 1928, and another article with him the following year on the Raman effect.
In 1930, Segrè began studying the Zeeman effect in certain alkaline metals. When his progress stalled because the diffraction grating he required to continue was not available in Italy, he wrote to four laboratories elsewhere in Europe asking for assistance and received an invitation from Pieter Zeeman to finish his work at Zeemans laboratory in Amsterdam. Segrè was awarded a Rockefeller Foundation fellowship and, on Fermis advice, elected to use it to study under Otto Stern in Hamburg. Working with Otto Frisch on space quantization produced results that apparently did not agree with the current theory; but Isidor Isaac Rabi showed that theory and experiment were in agreement if the nuclear spin of potassium was +1/2.