Glaser was born in Cleveland, Ohio, and took his degree in physics and mathematics at the Case Institute of Technology there. After graduating in 1946, he went on to gain his doctorate for cosmic-ray research from the California Institute of Technology in 1950. From 1949 to 1959, Glaser worked in the physics department of the University of Michigan, becoming professor in 1957. In 1959 he moved to the University of California at Berkeley as a professor of physics and subsequently (1964) as a professor of physics and biology.

While at the University of Michigan, Glaser became interested in techniques for the visualization and recording of elementary particles. The Wilson cloud chamber, using supersaturated vapor, had been in use since the 1920s, but was unsuited to the detection of the highly energetic particles emerging from the new accelerators of the 1950s.

Glaser considered other unstable systems that could be used, and experimented with superheated liquids, in which ionizing particles would leave a trail of vapor bubbles. In 1952 he produced the first radiation-sensitive bubble chamber, in which he used diethyl ether under pressure and controlled temperature. A sudden brief reduction in pressure was used and trails of bubbles forming along the tracks of particles could be captured by high-speed photography before the bulk of the liquid boiled. For this invention, and its subsequent development into a useful research tool, Glaser received the 1960 Nobel Prize for physics. At the University of California at Berkeley his interests turned to methods of applying physics to molecular biology.

The bubble chamber, using liquid hydrogen at low temperature, is now a basic component of almost all high-energy physics experiments, and has been the instrument of detection of many strange new particles and phenomena. Present-day bubble chambers are much bigger (and more expensive) than Glaser's original, which was only three cubic centimeters in volume.