In 1942, Dey accompanied Krishna Chandra Dey on a visit to Bombay. There he started working as an assistant music director first under Krishna Chandra Dey in a Bengali film Chanakya in 1939 and then under Sachin Dev Burman. Later, he assisted other music composers and then started to work independently. While working independently as a music director for various Hindi movies, Dey continued to take musical lessons in Hindustani classical music from Ustad Aman Ali Khan and Ustad Abdul Rahman Khan. He often works with near about 185 music directors in his singing career in Bollywood.

Dey started his career in playback singing with the movie Tamanna, in 1942. The musical score was by Krishna Chandra Dey and Manna sang a duet named "Jago Aayee Usha Ponchi Boley Jago" with Suraiya which was an instant hit.

But it was only in 1943 that he got his first solo break with Ram Rajya. Incidentally, the producer of the film Vijay Bhatt and its composer Shankar Rao Vyas had approached K C Dey with an offer for playback in the film. When K C Dey refused the offer on the grounds that he would not lend his voice to other actors, they spotted Manna Dey sitting in the corner of the room and offered him the opportunity.

Shankar Rao Vyas taught Manna Dey the songs and he chose to sing them in his uncle's distinct style. And thus started the illustrious career with the first song "Gayi tu gayi Seeta sati" (Ram Rajya, 1943).

His songs like "O Prem Diwani Sambhal Ke Chalna" from 1944 film Kadambari composed by Anil Biswas, "Dil Churaney Ki Liye from Dur Chaley" (1946) composed by Jafar Khurshid, his duets with Amira Bahee like "E Diniya Jara Suney" from Kamala (1946) and duet song "Aaj Bor Aayee" with Meena Kapoor from 1947 film Chaltey Chaltey became chartbusters in respective years. Between 1945 and 1947, many Dey-Rajkumari duets like "Hay Gagan Me Badal Tharey" in 1945 for the film Vikaramaditya, "Aowji Morey" from Insaaf (1946), all four duets from the film Geet Govind composed by Pandit Indra – "Kit Ho Nando Kumar", "Chorr Sakhi Aaj Laj", "Apney Hi Rang", "Lalit Labang Lata" from Geet Govind became popular.

He sang for first time songs composed by Sachin Dev Burman, Upar Gagan Vishal and Duniya Ke Logo in the 1950 movie Mashal, which became popular and from here his association with S.D.Burman began. Its lyrics were written by Kavi Pradeep. In 1952, Dey sang for a Bengali and a Marathi movie with the same name and storyline, Amar Bhupali. This established him as a leading playback singer in Bengali films and Marathi films as well by 1953.

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In December 1953, Dey married Sulochana Kumaran. She was originally from Kannur, Kerala. Together they had two daughters – Shuroma Herekar (1956–2016), a U.S. based scientist and Shumita Dev (b. 1958), a Bengaluru-based businesswoman. Sulochana died in Bengaluru in January 2012. She had been suffering from cancer for some time. After her death, Dey moved to Kalyan Nagar in Bengaluru after spending more than fifty years in Mumbai.

On 8 June 2013, Dey was admitted to the ICU in a Bengaluru hospital after a chest infection gave rise to other complications. His health gradually improved and about a month later doctors took him off the ventilator support. Later, he was discharged from the hospital.

He was hospitalized again in the first week of October 2013, and died of a cardiac arrest at 3:45 pm on 24 October at Narayana Hrudayalaya hospital in Bengaluru, aged 94. Musicians, politicians, cricketers and other notable persons issued statements on his death. He was cremated in Bengaluru itself.