It is well to first make a microscopical examination of the stool by taking one of the whitish epithelial flakes from the rice-water material and making a straight smear which is then dried and fixed with heat. This may be stained best by a dilute carbol fuchsin (1 to 10). Methylene blue makes a good stain, or more differential is that by Gram’s method which shows the Gram-negative spirilla stained by the bismarck brown counterstain, giving the appearance of fish parallel to one another in a stream. According to Koch a diagnosis can be made in this way of one-half the cases during an epidemic. The scintillating motility of cholera spirilla may strike one in the examination of the stool in hanging drop. Dunbar has a quick diagnostic method in which epithelial flakes from the stool are emulsified in peptone solution. Then on a slide, according to the method to be later described, is deposited a drop of 1 to 50 normal serum dilution and on the same slide a second drop of 1 to 500 dilution of cholera serum. A loopful of the suspected stool emulsion is rubbed up in each of these serum dilutions and we should have cessation of motility and clumping in the cholera immune serum provided the organisms in the stool are true cholera spirilla. In case of an autopsy on a suspected case of cholera one should tie off, between double ligatures, at least two 5-inch sections of small intestines, one just above the ileo-caecal valve and one taken from about the middle of the ileum. These portions of gut should be dropped into sterile salt mouth bottles, well stoppered and sent to a bacteriological laboratory as soon as possible. As the cholera spirilla, when associated with faecal bacteria, tend to die off within twelve to twenty-four hours it would probably be advisable to inoculate an agar or blood serum slant with material from the ileum at the same time the sections of gut are removed. For diagnosis of a cholera carrier with a normal stool or a cholera suspect with a diarrhoeal one inoculate 2 or 3 tubes of peptone solution with 2 or 3 loopfuls of material from the stool. With suspected carriers who are constipated and to whom one should not give purgatives we may insert into the rectum a rubber tube or a throat swab in order to obtain material immediately. The cholera spirilla grow rapidly and being strong aerobes, they grow on the surface of the fluid so that by taking a loopful from the surface, we may in three to eight hours obtain a pure culture. Should there be a pellicle present, this should be avoided in transfer by tilting the tube slightly, so that the material near the surface be obtained without touching the pellicle. Inoculate a second tube from the surface of this first and, if necessary, a third (_enrichment method_).