Hydrogen Chloride Gas for Effecting Staining of Internal Structures in Escherichia Coli.

The cells were smeared in water or water which had stood over about 10 mg. of magnesium powder per ml. for 30 minutes or longer. After the smear was dry and whitish in appearance it was held over a beaker of hot water (60-65° C.) until it was translucent or becoming translucent and exposed immediately to hydrogen chloride (gas) for a few seconds. After drying, it was covered with a 0.1% aqueous solution of neutral red for 5-8 minutes. The excess stain was washed from the slide with water and, while wet, placed in a saturated aqueous solution of mercuric nitrate for 5-15 seconds. The smear was rinsed in water and allowed to dry. When dry the slide was placed on a 50° C. warm plate and covered with a thin film of a 5% aqueous solution of nigrosin adjusted to a pH of about 3. The film dried quickly and upon cooling was ready for study. The stained material in the cells varied in shape and location with the moisture content of the smear and the time of exposure to hydrogen chloride. In the area of the smear directly exposed to the gas, the cells in general possessed a round or oval stained structure. Where there was little, if any, exposure to the gas the cells were uniformly stained. There were various gradations in the location and shape of the stained material in the cells from the one extreme to the other.