Madhabi Mukherjee was born on 10 February 1942, originally Madhuri Mukherjee. She was raised with her sister Manjari by their mother in Kolkata, in what was then Bengal, India. As a young girl, she became involved in the theatre.

She worked on stage with doyens like Sisir Bhaduri, Ahindra Choudhury, Nirmalendu Lahiri and Chhabi Biswas. Some of the plays she acted in included Naa and Kalarah. She made her film debut as a child artist in Premendra Mitra's Kankantala Light Railway (1950).

Madhabi first made a major impact with Mrinal Sen's Baishey Shravan (Wedding Day) in 1960. The film is set in a Bengal village just before and during the horrific famine of 1943 in Bengal that saw over 5 million die. Madhabi plays a 16-year-old girl who marries a middle-aged man. Initially she brightens up his life but then World War II and the Bengal Famine hits them. The couple's marriage disintegrates.

Her next major film was Ritwik Ghatak's Subarnarekha (The Golden Thread ) made in 1962 but released in 1965 – the last in a trilogy examining the socio-economic implications of partition, the other two being Meghe Dhaka Tara (The Cloud-Capped Star) (1960) and Komal Gandhar (E-Flat) (1961). It is also perhaps Ritwik Ghatak's most complex film. In the film Ghatak depicts the great economic and socio-political crisis eating up the very entrails of the existence of Bengal from 1948 to 1962 ; how the crisis has first and foremost left one bereft of one's conscience, one's moral sense. Madhabi gives a wonderful performance as Sita, the younger sister of Ishwar (Abhi Bhattacharya) who kills herself when as a prostitute waiting for her first customer, she finds out the customer is none other than her estranged brother.

In the early 1960s, she was recruited by Satyajit Ray to portray the role of Arati in the 1963 film, Mahanagar (The Big City).

Recalling her meeting with Ray, Madhabi wrote:

He read me the entire story, Mahanagar. I was stunned. This was the first woman-centered screenplay I had encountered. I was not going to play second fiddle to the main male character as in all plays and films I had acted in or was familiar with. (p.20)

In Mahanagar, Madhabi Mukherjee plays Arati who takes a job as a saleswoman due to financial constraints in the family. The large joint family is horrified at the thought of a working woman. For Arati, going door to door selling knitting machines opens up a whole new world and new friends and acquaintances including an Anglo-Indian friend, Edith. Earning money also raises Arati's status in the family especially when her husband (Anil Chatterjee) loses his job. When Edith is sacked unfairly, Arati resigns in protest...Madhabi's towering performance as Arati dominates the film. Quoting Film critic Roger Ebert: " it might be useful to see the performance of Madhabi Mukherjee in this film. She is a beautiful deep, wonderful actress who simply surpasses all ordinary standards of judgment."

This film was soon followed by her portrayals of Charu in Charulata (The Lonely Wife), the 1964 film based on Rabindranath Tagore's novella Nashtanir (The Broken Nest, 1901). Mukherjee's stunning portrayal of Charulata, a bored and neglected housewife of Calcutta in the 19th century, is undoubtedly a towering performance in the history of Indian cinema.

Without doubt Madhabi reached the peak of her career with this film, possibly Ray's greatest film as well, the Apu trilogy notwithstanding. As the bored and neglected housewife in Victorian Calcutta of the 1870s who gets attracted to her husband's cousin Amal (Soumitra Chatterjee), Madhabi makes the central role of Charu her own. It is without doubt one of the greatest performances of Indian cinema. She lives the role. She is Charulata. Till date Madhabi in Charulata remains the benchmark for what an ideal Tagore heroine should be and it is said that when Ray returned to Tagore with Ghare Baire (1984) (The Home and the World), he stylised Swatilekha Chatterjee in a manner similar to Madhabi in Charulata.

Madhabi's third and last film with Ray was Kapurush (The Coward) in 1965. The films looks at Amitabha Roy (Soumitra Chatterjee), a screenwriter whose car breaks down in a small town. He lodges with a local resident, Bimal Gupta (Haradhan Bannerjee). Bimal is married to Karuna (Madhabi Mukherjee), who was a past girlfriend of Amitabha, a fact which Bimal is unaware of. Despite playing out predictably, Kapurush still has a great deal of charm, most notably in the wordless acting prowess demonstrated by Soumitra and Madhabi. Through their subtle eye movements and small body gestures, we are able to discern their unspoken turmoil especially Madhabi's who behaves totally indifferently to Soumitra even as he tries to re-connect to her.

Though she remained a big star in the Bengali commercial film industry, after Kapurush Madhabi failed to reach such critical heights as her films with Ritwik Ghatak and Satyajit Ray again.

Her major films post Kapurush include Calcutta 71 in 1972 by Mrinal Sen, Biraj Bou in 1972 by Manu Sen, Strir Patra in 1972 by Purnendu Patri, Ganadevata in 1978 by Tarun Majumdar, Bancharamer Bagan in 1980 by Tapan Sinha, Chokh in 1982 then Chhandaneer in 1989 by Utpalendu Chakrabarty and Utsab in 2000 by Rituparno Ghosh.

Madhabi is married to Bengali film actor Nirmal Kumar. They have two daughters but are currently separated.

She wrote her autobiography Ami Madhabi in 1995.

• Bakita Byaktigato (2013)
• Hing Ting Chot (2010)
• Mayer Adar (2002)
• Utsab (2000)........ Bhagabati
• Rangin Basanta (1995)
• Artikram (1994)
• Prithibir Shesh Station (1993)
• Mon Mane Na (1993)...........Dipak's mother
• Daan Pratidaan (1992)
• Antarer Bhalobasha (1991)
• Agni Trishna (1989)
• Chandaneer (1989)
• Kari Diye Kinlam (1989)
• Aghaton Ajo Ghatey (1989)
• Anjali (1988)
• Ekti Jiban (1988)
• Pratikar (1987)
• Uttar Lipi (1986)
• Anurager Choa (1986)
• Bhalobasa Bhalobasa (1985)
• Putulghar (1985)
• Jog Biyog (1984)
• Samapti (1983)
• Chhoto Maa (1983)
• Chokh (1983)
• Matir Swarga (1982)
• Prafulla (1982)
• Subarnalata (1981)
• Manikchand (1981)............ Baroboudi
• Saheb (1981)
• Bancharamer Bagan (1980).............Chhakari's wife
• Ganadevata (1979).............Padma
• Yugo Manab Kabir (1976)
• Phool Sajya (1975)
• Natun Surya (1975)
• gnishwar (1975)
• Bindur Chheley (1973).............Bindubasini
• Bon Palashir Padabali (1973)
• Strir Patra (1972)
• Chinna Patra (1972)
• Biraj Bou (1972)
• Jiban Rahasya (1972)
• Calcutta 7 (1971)
• Chhadmabeshi (1971).............Sulekha
• Samantaral (1970)
• Duranta Charai (1969)
• Adwitiya (1968)
• Garh Nasimpur (1968)
• Chhotto Jignasa (1968)
• Kheya (1967)
• Shankhabela (1966)
• Swapna Niye (1966)
• Joradighir Chowdhury Paribar (1966)
• Kapurush (1965).............Karuna Gupta
• Ghoom Bhangar Gaan (1965)
• Subarnarekha (1965).............Sita
• Thana Theke Aschi (1965)
• Godhuli Belaye (1964)
• Binsati Janani (1964)
• Charulata (1964).............Charulata
• Mahanagar (1963)............. Arati Mazumder
• Aaj Kal Parshu (1961)
• Baishey Shravana (1960)
• Tonsil (1956)
• Kankantala Light Railway (1950)

• 2014:Winner, Filmfare East Lifetime achievement award
• 1970:Winner, National Film Award for Best Actress for-Dibratrir Kabya
• 1965:Winner, BFJA Award for Best Actress for -Charulata
• 1966:Winner, BFJA Award for Best Actress for -Dholgobinder Karcha
• 1967:Winner, BFJA Award for Best Actress Award for -Joradighir Chowdhury Paribar
• 1971:Winner, BFJA Award for Best Actress Award for -Dibaratir Kabya
• Kalakar Awards