A Safe Spore Stain for Class Use

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Abstract
In looking for an efficient method of pre-treating bacterial spores for staining with carbol fuchsin, the author found that the ammonia in Lagerberg's formula and the chromic acid in Moeller's formula were both fairly effective in increasing permeability. Combining the two proved highly successful; and the following method has been used for four years by classes of beginners in the study of spore-formers isolated from soil by the students. Cover the heat-fixed smear with five percent chromic acid and after thirty seconds add twice the amount of concentrated ammonia to the acid. After two minutes rinse and steam with carbol fuchsin for two or three minutes. Rinse, destain with one percent sulphuric acid for fifteen to thirty seconds, rinse again and flood slide with tap water. Add to this a few drops of Loeffler's methylene blue and allow to counter-stain for ten to thirty seconds. Rinse, blot, dry and examine. If instructions are followed the results are always good.

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