1886.] 



MICKOSCOPICAL JOUENAL. 



45 



2d. To 3 per cent, alum-water for 

 ten minutes. 



3d. To haematoxylin dye, diluted 

 with an equal part of 3 per cent, 

 alum-water, for one hour. 



4th. To full strength dye, if neces- 

 sary, for half or one hour. 



5th. To alum-water for a moment, 

 or until any excess of color is soaked 

 out. 



6th. Brush thoroughly in water, 

 and put into one ounce of clean water 

 for fifteen minutes, to remove alum 

 crystals. 



7th. To 93 per cent, alcohol for 

 fifteen minutes. 



8th. To absolute alcohol for two 

 hours, or longer. 



9th. To oil of cloves for one hour, 

 or until ready to mount. 



Some leaves, chiefly ferns with 

 sori, may be double-stained with 

 haematoxylin and anilin blue ; the 

 former going to sori and spirals, the 

 latter to other parts. The process is 

 first to stain in haematoxylin, and 

 then to soak the color in part from 

 the body of the leaf by putting it in 

 alum-water. Next carry through 

 pure water and alcohol to a half- 

 grain anilin blue solution for thirty 

 or forty-five seconds, and proceed as 

 you do with a single blue staining, 



II. DOUBLE STAINING OF SECTIONS. 



For double stainings I use hema- 

 toxylin and carmine, and bkie, green, 

 and red anilins. 



Of the red anilins I prefer that 

 known under the head of magenta 

 or roseine pure, though fuchsin, pon 

 ceau, and solferino may be used. 

 These anilins are manufactured at 

 the Atlas Works of Brooke, Simp- 

 son & Spiller, London. 



The anilin dyes are made by dis- 

 solving the quantity given in each 

 process with aid of mortar and pes- 

 tle, in one ounce of 93 per cent, al- 

 cohol and filtering. 



The haematoxylin and carmine 

 dyes are made according to the fol- 

 lowing formulge : 



Hcematoxylin Dye. 

 Ground Campeachj wood, h ounce. 

 Pulv. alum, I " 



Mix and tritui^ate in a mortar for 

 twenty minutes, then add five ounces 

 of hot distilled water, and let it 

 stand for two days. Filter, and to 

 each ounce of the dye add two 

 drachms of 75 per cent, alcohol. In 

 twenty-four hours again filter to re- 

 move precipitated alum. This dye 

 is made somewhat after Dr. Arnold's 

 formula ; he using the extract instead 

 of the wood. It keeps, with occa- 

 sional filterings, in w^ell-stoppered 

 bottles for two months. 



Borax Carmifie Dye. 



Pulv. carmine, . . . . 7^ grains. 



Saturated aqueous solu- 

 tion of borax, . . . 7^ A- dr. 



Mix and add absolute al- 

 cohol, 15 drachms. 



Filter and collect crystals when 

 dry. Dissolve nine grains of crystals 

 in one ounce of distilled water. 



This is Dr. J. J. Woodward's for- 

 mula ; but not so strong, as his is a 

 saturated solution. 



Ammonia Carmine. 

 Pulv. carmine, . . . . 7^ grains. 

 Water of ammonia, . . 20 drops. 

 Absolute alcohol, ... h ounce. 



Gh'cerin, i " 



Distilled water, ... i " 



Put the pulverized carmine in a 

 test-tube, and add the ammonia. 

 Boil slowly for a few seconds, and 

 set aside, uncorked, for a day, to get 

 rid of excess of ammonia. Add the 

 mixed water and glycerin, and next 

 the alcohol ; then filter. 



Process I. — To Stain Sections with 

 Magenta and Blue Anilin. 



I St. Transfer froin alcohol to ma- 

 genta dye (one quarter of a grain to 

 the ounce) , and let it remain from 

 fifteen to thirty minutes. 



2d. Soak in alcohol for about the 

 same time, or until the color is en- 

 tirely, or in great part, removed from 

 parenchymal tissue. 



3d. Place or hold in a quarter or 

 a half-grain anilin blud" solution from 

 fifteen to forty-five seconds. 



