Staff and patients at Standish Hospital near Stroud are in the final stages of moving to a new site.
The future of the old building is under consideration
Tuesday is the last day of operation before the entire facility is relocated to Gloucestershire Royal Hospital.
Ambulances will ferry patients to the new premises, with nurses accompanying them where possible.
The move is expected to take two days and relatives are asked to wait until 6pm on the 29th and 30th to visit so people can settle into the new wards.
The move brings to an end years of protests by local residents to keep the historic hospital open.
The building, formerly a stately home, was given to the Red Cross a century ago.
It was put into service as a military hospital, treating TB cases.
More recently, it has specialised in orthopaedics, rheumatology and respiratory care.
The future of the old building now depends on two bids currently under consideration by Gloucestershire NHS Trust.