Only 200 people, chosen on a first come, first served basis, will be able to take the trip and receive a special souvenir from the State Railway of Thailand (SRT).
Tickets are priced at 1,499 baht per person, covering breakfast, lunch, snacks and beverages.
Tickets are now available at all SRT train stations, including the online channel.
The Kiha 183 diesel-powered train will leave Hua Lamphong Station in Bangkok at 8.10am on Saturday and return at 7.45pm.
Passengers will be able to visit Chao Sam Phraya National Museum, Wat Chaiwatthanaram, Ayutthaya Historical Park, Wat Phanan Choeng and Kong Khong Market.
They will be able to visit Bang Pa-in station, a place where King Rama V arranged lunch for vassals and diplomats during the opening of Thailand's first railway from Bangkok to Ayutthaya on March 26, 1896.
"Passengers will be able to take the Kiha 183 diesel-powered train ride to experience the art and culture along the Bangkok-Ayutthaya route," SRT public relations director Ekarat Sriarayanpong said.
He advised people taking the trip to wear Thai traditional dress.
Ekarat said the train trip was a cooperation between the SRT, the Tourism Authority of Thailand and related agencies to stimulate domestic tourism and generate income for locals.
He added that the SRT is planning train trips to many provinces every Saturday and Sunday to meet the needs of tourists.