. Pathogenic microörganisms; a practical manual for students, physicians, and health officers . l the acidIS replaced; (3) wash in 1 or 2 per cent, solution of sodium chloride and mountin the same. Do not use water at any stage. The capsule stains a pale violet.(See Plate III, Fig. 15.) Hisss Copper Sulphate Method (Fig. 13).—The organisms are grown,jf possible, on ascitic fluid or serum media. If not, the organisms should be MICROSCOPIC METHODS 79 spread on the cover-glass mixed with a drop of serum, or better, with a dropof one of the diluted serum media. Dry in the air and fix by heat. The
. Pathogenic microörganisms; a practical manual for students, physicians, and health officers . l the acidIS replaced; (3) wash in 1 or 2 per cent, solution of sodium chloride and mountin the same. Do not use water at any stage. The capsule stains a pale violet.(See Plate III, Fig. 15.) Hisss Copper Sulphate Method (Fig. 13).—The organisms are grown,jf possible, on ascitic fluid or serum media. If not, the organisms should be MICROSCOPIC METHODS 79 spread on the cover-glass mixed with a drop of serum, or better, with a dropof one of the diluted serum media. Dry in the air and fix by heat. The capsules are stained as follows: A 5 per cent, or 10 per cent, aqueoussolution of gentian violet or fuehsin (5 satm-ated alcohoUc solution gentianviolet to 95 distilled water) is used. This is placed on the dried and fixedcover-glass preparation and gently heated for a few seconds until steam dye is washed off with a 20 per cent, solution of copper sulphate (crystals).The preparation is then placed between filter paper and thoroughlv dried(Plate III, Figs. 14 and 10).. Fig. 13.—Capsule stain by Hisss method. Rhinoscleroma bacillus. X 1000. (Thro.) Huntoons Method (Fig. 1, p. 33).—^A 3 per cent, solution of nutrose iscooked for 1 hour in an Arnold and tubed unfiltered after adding per cent,carbohc. The organisms to be stained are mixed with a drop of nutrose, spreadin thin film on glass shde and dried in air, not fixed. The stain is made up per cent, concentrated nitric acid, 1 per cent, of a 1 per cent, acetic acid solu-tion, 1 per cent, of alcoholic solution of basic fuehsin (or other stain), and 1 percent, of carbol-fuchsin all in distiUed water. The stain is kept on the film for30 seconds, washed in water, and dried. Staining Spores and Acid-fast Bacteria.—^We have already riotedthat during certain stages in the growth of a number of bacteria, sporesare formed which refuse to take up color when the bacteria are stainedin the ordinary
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