Many theories have been proposed to explain the acid-fast property of the tubercle bacillus, but no satisfactory explanation has yet been given. It is suggested that the property is dependent upon the permeability of the cytoplasmic membrane. Evidence will be presented in support of this concept. It will be shown that when the Ziehl-Neelsen technique is employed the dye exists within the cell in two distinct portions: a small portion is bound to the cytoplasm and the remainder is free. The characteristic color of the stained bacillus is due to the free dye which can be removed without altering the acidfast believed that this phenomenon of acidfastness might be based upon the properties of chemicals isolated from the tubercle bacillus or of complexes of these chemicals. At one time there was a general belief that the presence of a wax sheath around the cell was responsible for its peculiar staining characteristics, but this is no longer tenable after the study made by Knaysi (1929). Some authors including Ehrlich thought that acid-fastness was related to the permeability of the cell membrane, but this structure was not clearly defined and experimental evidence was not given. The one point on which there is complete agreement is that the integrity of the cellular structure must be maintained to preserve the acid-fast property.Since our proposed explanation of the mechanism of acid-fastness is based on the function of a cellular structure, a brief review of pertinent information on the cytology of the tubercle bacillus is indicated.The cell wall of the tubercle bacillus has been observed directly with the electron microscope by Mudd and Anderson (1944). Mudd and Mudd (1927) demonstrated the hydrophobic property of the surface of the bacillus. The apparent functions of the cell wall are to protect the cell from mechanical injury and to impart to the cell its characteristic shape. The electron microscopic studies of Mudd, Polevitsky, and Anderson (1942) and Mudd and Anderson (1944) give direct evidence of the cytoplasmic membrane in bacteria. According to Knaysil (1929Knaysil ( , 1938Knaysil ( , 1944Knaysil ( , 1946, who refers frequently to this structure, the membrane of the tubercle bacillus probably consists of lipids and protein. With ordinary technique it appears to be the external boundary of the cytoplasm rather than a separate strtucture.
EXPERIMENTAL RESULTSAlthough there is some evidence that the cell wall of the tubercle bacillus retains small amounts of certain other dyes, our observations suggest that this 1 A recent personal communication from Knaysi states that what he referred to as the cell "membrane" in his paper (1929) is now usually known as the cytoplamic membran. 777