ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 903 



litre of aq. dest. In this the sections are left for 24 hours. Then 

 wash and transfer to an alcoholic solution of eosin (as in No. 4) for 

 several minutes. Wash in alcohol, and clarify. This method pro- 

 duces a beautiful nuclear stain, the nuclei are coloured deep violet, 

 the rest of the tissue a beautiful rose red. 



6. Hsematoxyliu-glycerin and rosanilin nitrate. The sections are 

 placed in the dilute hsematoxylin-glycerin (No. 5) for 24 hours ; 

 then for ten minutes in a solution of rosanilin nitrate ; after washing 

 in water, dehydrate and clarify. 



7. Methyl-green and rosanilin nitrate. The sections are left in 

 the No. 1 solution of methyl-green for ten minutes, then after washing 

 are placed in solution of rosanilin nitrate for fifteen minutes ; wash 

 again, dehydrate, clarify. 



The first three methods may be modified by using very dilute 

 solutions and staining for 24 hours. For hardening, the author 

 always employed absolute alcohol ; and Miiller's fluid or chromic acid 

 for material to be imbedded in celloidin. 



Staining Nerves in Muscle. * — To obtain good images of the 

 ramification of nerves in muscle. Dr. K. Mays gives the following 

 process. For small muscles, lay freshly prepared portions in a recently 

 made mixture of 1/2 per cent, alkaline solution of gold chloride, 

 1 grm. ; 2 per cent, osmic acid, 1 grm. ; water, 20 grm., until the arbo- 

 rescent nerve ramifications can be perceived ; then in the following 

 mixture : glycerin, 40 grm. ; water, 20 grm. ; 25 per cent, hydro- 

 chloric acid solution, 1 • grm., for about a day. This prevents them 

 from becoming too dark. 



Thicker muscles are placed for 12 hours in a 2 per cent, solution 

 of acetic acid, then into a freshly made mixture of 1/2 per cent, 

 alkaline solution of gold chloride, 1 grm. ; 2 per cent, osmic acid, 

 1 grm. ; 2 per cent, acetic acid, 50 grm. In this they remain for two 

 or three hours and are then placed in the glycerin mixture. The 

 muscle becomes transparent and amber-coloured, the nerve black- 

 brown. By this method the nerve-end-plate is stained, but not the 

 hypolemmal parts of the nerve. 



Anilin-gTeen.| — Dr. J. H. List controverts Schiefferdecker's 

 statement that anilin-green requires exposure to light before it is 

 fully capable of staining cell-structures. He finds that solutions of 

 methyl-green and anilin-green (0'5 grm.-lOO c.c. distilled water) 

 always colour, even when freshly prepared, the reticulum of goblet 

 and other cells. He also recommends Bismarck-brown and rosanilin 

 nitrate, either in union or alone for the same object. 



In reply to List, Dr. P. Schiefferdecker J maintains that the 

 former has confused the reticulum which is perfectly apparent even 

 in unstained specimens, with the reticulum only demonstrable by 

 anilin-green solution which has undergone certain changes from lapse 

 of time and exposure to light. The latter reticulum is much thicker 

 than that which is described by List, but otherwise there does not 

 seem to be much difference between them. 



• Ztitschr. f. Biol., xx. (1884) pp. 4 19-528 (5 pis.). 



t Zeitachr. f. Wias. Mikr., ii. (1885) pp. 222-3. X Ibid-, pp. 223-4. 



3 N 2 



