462 SU.MMARY OF CURRENT RESEARCHES RELATING 10 



method has been used with success for many Invertebrates, and many 

 mammalian and human tissues, which tend to disintegrate into fragments 

 with the usual freezing methods. 



C4} staining' and Injecting. 



New Method of Staining Diphtheria Bacilli.* — C. Ponder has 

 found that the following procedure is extremely satisfactory for detect- 

 ing Bacillus diphtherise in direct smears and in films from cultures. A 

 film is made on a cover-glass and fixed in the usual way. It is then 

 treated with the following stain: — Toluidin-blue 0'02 grm. ; glacial 

 acetic-acid 1 c.cm., absolute alcohol 2 c.cm., distilled water to lOU c.cm. 

 A loopful of the fluid is dabljed on and spread over the film. The cover- 

 slip is then turned over and mounted as a hanging drop. It is then 

 examined by artificial light. Diphtheria bacilli are stained a pale blue, 

 while the granules are of a deep red hue. The rings for the hanging drop 

 are made with plasticin. 



Demonstrating Unstriped Muscle Fibres in the Intima of the 

 Human Aorta. t — A. Ollendorff alludes to the difficulty of demonstrating 

 the presence of unstriped muscle fibres in blood vessels, especially if they 

 be isolated or in small bundles. He adopted Benda's procedure for 

 staining the myoglia or supporting substance of the smooth musculature. 

 The material used was obtained from bodies 20 to 25 years of age and 

 removed not later than 2 to 3 hours after death, fresh tissue being of 

 the greatest importance. The material was at once placed in Zenker's 

 fluid for 24 hours and then w'ashed for several hours in water. The 

 third step was to make frozen sections, which were immersed for 24 hours 

 in 0*5 p.c. chromic acid solution. After a wash in water the sections 

 were treated for ;-i minutes with 0'25 p.c. solution of permanganate of 

 potash. After another wash in water they were removed to Pal's mixture 

 of sulphite of soda and oxalic acid for 5 minutes. After another wash 

 in water the sections were placed on slides and stained with Benda's 

 mixture (crystal violet, hydrochloric acid, alcohol and anilin water). The 

 stain is mopped up with blotting paper and then dilute Lugol's solution 

 poured over the section. Next, the Lugol is mopped off, the section 

 dried, and then differentiated with a mixture of equal parts of anilin-oil 

 and xylol. After this it is treated with xylol and mounted in balsam. 

 The muscle fibres whose margins are not visible are seen as violet-coloured 

 streaks, while the nuclei are quite evident. 



Demonstrating the Structure of Microfilarise.J — G. Pittaluga finds 

 that the best way to show the structure of filarial is to adopt the intra- 

 vitam method of staining. The material used was obtained from in- 

 habitants of Spanish G-uinea. A small drop of live blood from the ear 

 or finger is placed on a slide and mixed with a similar or suitable quan- 

 tity of a solution of alkalin-methylen-blue. 



Simple Methods in the Bacteriological Diagnosis of Cholera.§ — 

 C. Krumwiede read a paper before the Society of American Bacterio- 



* Leanest (1912) pp. 22-3. 



t Anat. Anzeig. xxxviii. (1911), pp. 569-73. 



i Bol. Instit. Nacional Higiene Alfonso XIII. viii. (1912) pp. 77-92 (3 pis.). 



§ Centralbl. Bak.t., Ite Abt. Ref., li. (1912) pp. 685-6. 



