Santiago Felipe Ramon y Cajal investigated brains in the nineteenth and twentieth centuries in Spain. He identified and individuated many components of the brain, including the neuron and the axon. He used chick embryos instead of adult animals, then customary in brain research, to study the development and physiology of the cerebellum, spinal cord, and retina. Ramon y Cajal received the Nobel Prize in Physiology and Medicine in 1906, along with Camillo Golgi, for his work on the structure of the nervous system.

The Golgi staining technique, also called the black reaction after the stain's color, was developed in the 1870s and 1880s in Italy to make brain cells (neurons) visible under the microscope. Camillo Golgi developed the technique while working with nervous tissue, which required Golgi to examine cell structure under the microscope. Golgi improved upon existing methods of staining, enabling scientists to view entire neurons for the first time and changing the way people discussed the development and composition of the brain's cells. Into the twenty-fist century, Golgi's staining method continued to inform research on the nervous system, particularly regarding embryonic development.