Madhabi Mukherjee was born in 1943. Her original name was Madhuri. She started acting at a young age as child artist. She made her film debut as a child artist in Premendra Mitra's Kankantala Light Railway (1950). First acted in a lead female role in Tapan Sinha's Tonsil (1956). The next film she worked on was Mrinal Sen's Baishey Shravan (1960); Bijoy Chatterjee, the producer of this film gave her the name Madhabi. After appearing in Mrinal Sen's film, Madhabi acted in another maestro Ritwik Ghatak's 'Subarnarekha'.
She created remarkable impact with her performance as Charulata in Satyajit Ray's film 'Charulata' based on Rabindranath Tagore's novel. She also displayed memorable performance in Ray's 'Mahanagar' and 'Kapurush'.
Madhabi acted as a heroine and in character role with many actors. She did some performance on stage and directed one Bengali movie (Aatmaja).