Tyndall was born into a poor Protestant Irish family. After a thorough basic education he worked as a surveyor in Ireland and England (1839–47). When his ambitions turned from engineering to science, Tyndall spent his savings on gaining a Ph.D. from the University of Marburg, Germany (1848–50), but then struggled to find employment.

In 1853 Tyndall was appointed professor of natural philosophy at the Royal Institution, London. There he became a friend of the much-admired physicist and chemist Michael Faraday (Faraday, Michael), entertained and instructed fashionable audiences with brilliant lecture demonstrations (rivaling the biologist T.H. Huxley (Huxley, T.H.) in his popular reputation), and pursued his research. An outstanding experimenter, particularly in atmospheric physics, Tyndall examined the transmission of both radiant heat and light through various gases and vapours. He discovered that water vapour absorbs much more radiant heat than the gases of the atmosphere and argued the consequent importance of atmospheric water vapour in moderating the Earth's climate—that is, in the natural greenhouse effect. Tyndall also studied the diffusion of light by large molecules and dust, known as the Tyndall effect, and he performed experiments demonstrating that the sky's blue colour results from the scattering of the Sun's rays by molecules in the atmosphere.

Tyndall was passionate and sensitive, quick to feel personal slights and to defend underdogs. Physically tough, he was a daring mountaineer. His greatest fame came from his activities as an advocate and interpreter of science. Tyndall, in collaboration with his scientific friends in the small, private X Club, urged greater recognition of both the intellectual authority and practical benefits of science. He was accused of materialism and atheism after his presidential address at the 1874 meeting of the British Association for the Advancement of Science, when he claimed that cosmological theory belonged to science rather than theology and that matter had the power within itself to produce life. In the ensuing notoriety over this “Belfast Address,” Tyndall's allusions to the limitations of science and to mysteries beyond human understanding were overlooked. Tyndall engaged in a number of other controversies—for example, over spontaneous generation, the efficacy of prayer, and Home Rule for Ireland.