Observations of the Golgi Substance with the Electron Microscope

Abstract

OBSERVATIONS by several investigators during the past few years lend support to the view that the Golgi substance is present in living cells in the form of droplets stainable supra-vitally with certain vital dyes^1–4. The classical form of the Golgi substance has been considered to be a gross artefact induced by the development of myelin figures from droplets of phospholipid^3,4. This view has been accepted by some^5–7 and questioned by others^8,9. It was felt that observations with the electron microscope might aid in the elucidation of this complex problem. With this possibility in mind, liver and intestinal epithelium were fixed by perfusion in two different fixatives, one the Kolatchev–Nassonov modification of Champy's fluid¹⁰ and the other a mixture of equal parts of 2 per cent osmic acid, 3 per cent potassium dichromate and 2 per cent lanthanum nitrate. The pH of the former fixative is 2.0 and that of the latter is 3.4. After fixation, slices of tissue were transferred to the same fixative for varying lengths of time. Both tissues were examined during the period of fixation with oil immersion at a thousand diameters by crushing the tissues under a coverslip in water or glycerine. Neither myelin figures nor the Golgi substance could be identified during the first twenty-four hours with the osmic acid, potassium dichromate, lanthanum nitrate mixture, nor during the first forty-eight hours fixation with modified Champy's fluid. Further treatment with 2 per cent osmic acid for three days at 37° C. resulted in the appearance of the Golgi substance with its typical form and distribution throughout both tissues after fixation in modified Champy's fluid but not after fixation in the osmic, dichromate, lanthanum mixture. Secondary treatment of the tissues preserved in the latter fixative with a mixture of equal parts of 3 per cent potassium dichromate and 1 per cent chromic acid followed by treatment with 2 per cent osmic acid for four days at 37° C. resulted in the development of faint positive images of the Golgi substance, whereas secondary treatment with modified Champy's fluid for twenty-four hours followed by treatment with 2 per cent osmic acid at 37° C. for four days resulted in the development of typical, well-blackened Golgi figures in both tissues.