Eisenstaedt's most famous photograph is of an American sailor kissing a young woman on August 14, 1945 in Times Square. (The photograph is known under various names: V-J day in Times Square, V-Day, etc.) Because Eisenstaedt was photographing rapidly changing events during the V-J Day celebrations, he stated that he didn't get a chance to obtain names and details, which has encouraged a number of mutually incompatible claims to the identity of the subjects.

Eisenstaedt was successful enough to become a full-time photographer in 1929. Four years later he photographed a meeting between Adolf Hitler and Benito Mussolini in Italy. Other notable pictures taken by Eisenstaedt in his early career include a waiter ice skating in St. Moritz in 1932 and Joseph Goebbels at the League of Nations in Geneva in 1933. Although initially friendly, Goebbels scowled for the photograph when he learned that Eisenstaedt was Jewish.

Because of oppression in Hitler's Nazi Germany, Eisenstaedt emigrated to the United States in 1935, where he lived in Jackson Heights, Queens, New York, for the rest of his life. He worked as a photographer for Life magazine from 1936 to 1972. His photos of news events and celebrities, such as Dagmar, Sophia Loren and Ernest Hemingway, appeared on 90 Life covers.

Eisenstaedt, known as "Eisie" to his close friends, enjoyed his annual August vacations on the island of Martha's Vineyard for 50 years. When on assignment in the Galapagos Islands, Eisenstaedt left the Galapagos prior to the assignment's completion so he could arrive on time for his Vineyard vacation in the Menemsha area of the town of Chilmark.[citation needed] During his Vineyard summers, he would conduct photographic "experiments," by working with various lenses, filters, and prisms, but always working with natural light. Eisenstaedt was fond of Martha's Vineyard's photogenic lighthouses, and was the focus of lighthouse fund raisers for the Vineyard Environmental Research Institute (VERI), the lease-holder of the lighthouses. One fund raiser was titled "Eisenstaedt Day" and was an international event. The last Eisenstaedt lighthouse fundraiser was held in August 1995, the month of his death on Martha's Vineyard.
He was awarded the National Medal of Arts in 1989.[5]
Eisenstaedt's last photographs were of President Bill Clinton with wife, Hillary, and daughter, Chelsea, on August 1993, at the Granary Gallery in West Tisbury on Martha's Vineyard. This historic "private" photo-session took place in a fenced-in courtyard protected by the Secret Service for over one hour, and was fully documented by William E. Marks.Marks, who took hundreds of photographs of Eisenstaedt in every situation imaginable for over ten years, also photographed Eisenstaedt signing his famous V-J Day photograph on the morning before his death.