Evolution of staining methods in neuroanatomy: Impetus for emanation of neuron doctrine during the turn of 20th century

The nervous system is distinctive as compared to other tissue systems in human body owing to intricate structural organization. Histological studies played a key role in unveiling complex details of nervous tissue. However, the process of developing suitable staining method for nerve cells was arduous and spanned across almost half a century. The present study explored details of the journey involving quest for propitious staining method in neuroanatomy culminating in promulgation of neuron doctrine at the onset of 20th century. Initial efforts involving hematoxylin (including its diverse modifications) and subsequent adoption of analogous dye-based stains (like Nissl's method) had limited success in visualization of different parts of a nerve cell and structural details of nervous tissue. This was due to inability of dye-based stains to penetrate the connective tissue sheath of nervous tissue. Eventually, advent of metallic stains in form of silver impregnation method (Golgi stain), reduced silver impregnation method with gold stain (Cajal's stain) and silver carbonate staining method of Río-Hortega unraveled the structure of nervous tissue. The evolution of staining methods catalyzed the refinement of theories pertinent to constitution of nervous tissue. Golgi's staining led to emergence of reticular theory (neurons exist as a network) and Nissl's staining was the basis of the concept of Nervösen Grau (nerve cells and glial cells are embedded in mass of gray matter). Finally, Cajal's staining method successfully elucidated the complex anatomy of nerve terminals and resulted in emanation of neuron doctrine (neurons exists as individual units with adjacent connections).