ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 503 



C4) Staining: and Injecting-. 



Staining Tubercle Bacilli.* — H. Biersy describes the following new 

 procedure for staining tubercle bacilli in sputum : — 1. To 1 vol. of 

 sputum add 2 vols, of distilled water, if the sputum be thin ; but if thick, 

 5 vols, of distilled water. Then add hypochlorite soda 1 drop plus 

 20 to 40 drops of 1 p.c. caustic soda to 5 c.cm. of sputum. 2. , Heat the 

 mixture at a temperature not exceeding 37°. Add soda solution drop by 

 drop until the mixture is quite homogeneous. ;>. Pour the mixture into 

 a test tube with rubber stopper (1 vol. plus 1 vol. of distilled water). 

 Mix. Add drop by drop acetic acid until the reaction is feebly acid to 

 litmus. Centrifuge for ten minutes ; decant off supernatant and reserve 

 it. Spread the deposit on slides. Add acetic acid to the decanted fluid 

 until a precipitate forms. Centrifuge, and then spread the new deposit 

 on slides. Make films of the deposit on slides ; dry, fix, and stain by 

 Ziehl-Xeelsen method. 



Staining with Alizarin. f — G. Colosl says the following method 

 gives good results. The material used was chiefly Invertebrates. The 

 material was fixed and embedded in paraffin. The sections were bathed 

 for a few seconds in a 1 p.c. aqueous solution of acetate of uranium. 

 After washing in running water, the sections were treated with 

 1 to 200 aqueous sulphate of soda and alizarin. After a few seconds 

 the preparations were washed, and, having been passed through up- 

 graded alcohols, were mounted. The medium does not seem to be of 

 importance. 



New Technique for Staining Sections with HaBmalum-eosin.l 

 L. Tribendeau recommends the following method of preparing and 

 employing solutions of hajmalum-eosin : 



Preparation oj Mother -solution. — Place 1 grm. of nitrate of silver 

 crystals in a clean corked flask of some 150 c.cm. capacity, and which 

 has been rinsed previously with distilled water ; add 100 c.cm. of 

 distilled water and dissolve by shaking. Into this solution empty as 

 quickly as possible 50 c.cm. of solution of potash (oxide of potash pure 

 10 grm., distilled water 100 c.cm.), cork the flask immediately, and shake 

 it for some seconds while holding it by the neck and the cork. Allow 

 it to stand for about one minute, and then decant off the cloudy and 

 brownish supernatant fluid down to the level of the heavy precipitate of 

 oxide of silver which collects at the bottom of the flask. Add 150 c.cm. 

 of distilled water to this precipitate, cork, shake again, set aside for a 

 minute, and re-decant as above. Repeat the whole process three or four 

 times. 



Dissolve 2 • 5 grm. of htematoxylin (f rangaise) in 50 c.cm. of absolute 

 ethyl-alcohol. Add this solution to the oxide of silver precipitate 

 and empty the mixture into a long-necked flask, place in the water- 

 bath, and shake from time to time until the alcohol starts to boil. 



* Comptes Rendus, clxiii. (1916) pp. 110-2. 

 t Monitore Zool. Ital., xxvi. (1915) pp. 248-51. 

 X C.R. Soc. Biol. Paris, Ixxis. (1916) pp. 288-9. 



2 M 2 



