The crew was reduced to a single pilot; the 64-foot wingspan was increased to 105 feet; more powerful Pratt & Whitney J57 engines replaced the J65s and all but the first six planes were equipped for air refueling.

Missions included Sky Shield, Toy Soldier and Green Hornet which were classified surveillance programs, which later would be performed by the U-2's of the 4028th SRS.

Between March and August 1958 the RB-57Ds Flew air sampling flights from the Marshall Islands, Eniwetok Proving Ground in conjunction with the USA Nuclear testing program (Code name Hardtack).

As a result of the 1963 Partial Nuclear Test Ban Treaty, the HASP program had been terminated, so all U-2 aircraft had the HASP air sampling equipment removed and were refitted with photo and electronic (PHOTINT & ELINT) collection gear The 4080th SRW then moved to Laughlin AFB, Texas on 1 April 1957 and was designated the 4080th Strategic Wing on 15 June 1960.

Despite Soviet Premier Nikita S. Khrushchev's denial, President John F. Kennedy directed SAC to begin U-2 high-altitude reconnaissance flights over the island.

After high-altitude reconnaissance flights over Cuba, Majors Richard Heyser and Rudolf Anderson returned with pictures of ballistic missile sites and nuclear storage facilities under construction.

President John F. Kennedy addressed the nation 22 October and six tense days followed while Heyser, Anderson and several other U-2 pilots from the 4080th continued reconnaissance flights over Cuba from a forward operating location established at McCoy AFB, Florida for the duration of the crisis.

During this move they had a small group of people assigned/attached who were working on the classified programme "Lightning Bug" which were photo reconnaissance drones.

On 31 December 1963 President Lyndon B. Johnson issued the order for the 4080th SRW to support the reconnaissance effort in South Vietnam.

They moved a detachment of three 4028th SRS U-2E aircraft (#347, #370 & #374) to Bien Hoa Air Base to supplement the tactical reconnaissance being undertaken by the F-101s.

The aircraft were quickly moved to Andersen AFB in Guam, and finally after some weeks of inactivity to Bien Hoa as OL-20 on 5 March with operations starting also most immediately.

The 4080th was a SAC major command controlled (MAJCON) wing, and in order to retain the lineage of its MAJCOM 4-digit combat units and to perpetuate the lineage of many currently inactive bombardment units with illustrious World War II records, Headquarters SAC received authority from Headquarters USAF to discontinue its MAJCON wings that were equipped with combat aircraft and to activate inactive Air Force controlled (AFCON) units, most of which were inactive at the time which could carry a lineage and history.

As a result, the 4080th SRW designation was discontinued and its mission, equipment and personnel transferred to the 100th Strategic Reconnaissance Wing in June 1966.