ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 635 



(4) Staining- and Injecting-. 



Solution for Staining Protozoa and Blood-corpuscles.* — T. 

 Watabiki makes the solutions in the following way: — Solution 1 : 

 methyl-blue (sic) 1 ; absolute alcohol, 10 ; carbonate of soda, 1 ; distilled 

 water, 90. This mixture is incubated for two days at 37° C. Solution 2 

 consists of yellow water-soluble eosin, 1 ; distilled water, 200. The 

 two solutions are mixed, and then incubated for twenty-four hours 

 at 37° C. The dry powder, • 3-0 • 5 grm., is called methylen-azureosine. 

 The final solution .consists of methylen-azureosine, • 5 ; methyl-alcohol, 

 150 ; neutral glycerin, 150. Staining of preparations is done in the 

 usual way. Leucocytes and bacteria stain in three to five minutes ; 

 Protozoa take from five to thirty minutes. 



Demonstrating the Presence of Spirochaetes in the Urine in 

 Cases of Trench Fever.f— A. T, Nankivell and C. E. Sundell adopted 

 the following procedure : — Ten c.cm. of urine are centrifuged at high 

 speed for fifteen minutes. The supernatant fluid is pipetted off and 

 5 c.cm. of distilled water added. The fluid is again centrifuged for 

 fifteen minutes and the supernatant fluid pipetted off. Films are then 

 made by placing three loopfuls on a slide. The preparation is then 

 dried in an incubator. The majority of the specimens were stained by 

 a modified Fantana silver method. The film is washed for two minutes 

 with a solution of 8 p.c. formalin and 1 p.c. glacial acetic acid. The 

 film is then washed in absolute alcohol, and when nearly dry is washed 

 with distilled water and then flooded with a hot 5 p.c. solution of tannic 

 acid, containing 1 p.c. of carbolic acid. After allowing the mordant to 

 act for two minutes the film is washed in distilled water. A 2 p.c. silver 

 nitrate solution, to which a trace of ammonium hydrate is added, is 

 pipetted over the film until a dark brown colour is obtained. The slide 

 is then washed with distilled water and afterwards dried in a current of 

 cold air. The film should be mounted in Canada balsam. The spiro- 

 chaetes appear jet black against a grey or brown background. With 

 dark ground illumination they are white. With Giemsa's stain they 

 stain pale red with a tinge of blue, but with dark ground illumination 

 they are yellow. An alternative staining to Fontana or Giemsa is 

 carbol-fuchsin. The spirochsetes stain pink by this method. 



(6) Miscellaneous. 



Micro-chemical Reaction for Calcite.$ — St. J. Thugutt states that 

 calcite when intermixed with zeolites can be recognized by the following 

 reactions : The powdered mineral (grains about 0"1 mm. diam.) is heated 

 on platinum foil over a Tecla burner for fifteen seconds and then treated 

 with N/10 cobalt nitrate. The calcite becomes coated with a thin skin 

 of blue basic cobalt salt, whilst the zeolites are. unaffected. Removing 



* Kitasato Archives of Eiper. Med., i. (1917) pp. 153-6. 



t Lancet (1917) ii. pp. 672-4. 



X Journ. Chem. Soc, ii. (1917) p. 508, 



2 u 2 



