Journal of Applied Microscopy. 583 



The society was indebted to Ohio State University for many courtesies 

 extended to it, and to the local committee, under the chairmanship of Dr. A. 

 M. Bleile, for the very satisfactory arrangements made to insure the success of 

 the meeting. The trolley ride under the guidance of Mr. Feiel, another member 

 of the local committee, was also thoroughly enjoyed. Henry B. Ward. 



University of Nebraska. 



Methods of Staining. — The following methods are given by request : 

 Stroebe's anilin blue stain for nerve fibers in hardened sections. — -Harden the 

 tissue in Miiller's fluid ; stain one-half to one hour in a saturated aqueous solu- 

 tion of anilin blue ; wash in water ; transfer to a small dish of water containing 

 twenty to thirty drops of a one per cent, alcoholic solution of caustic potash, 

 (potassium 1 ; alcohol 100 ; let the mixture stand twenty-four hours and filter). 

 In from one to several minutes the sections become bright brownish-red and 

 transparent ; place in distilled water five minutes ; sections turn blue ; stain in a 

 half-saturated solution of safranin for a quarter to one-half hour ; wash ; dehy- 

 drate in absolute alcohol, and mount. 



Unna'' s polychrome methylen blue solution: 



Methylen blue, 1 part. 



Carbonate of potassium, 1 part. 



Distilled water, 100 parts. 

 This solution is oxidized for several months, when methyl violet and methyl red 

 are formed. The ripened solution is sold by Griibler. 



Unna^s modified orcein method: 



Griibler's orcein, 1 part. 

 Hydrochloric acid, 1 part. 

 Absolute alcohol, 100 parts. 

 The sections are put in a porcelain capsule with just enough of the stain to 

 cover them, and warmed to about 30 degrees C. In ten or fifteen minutes the 

 stain becomes thick, owing to the evaporation of the alcohol. The sections are 

 then rinsed in alcohol, cleared and mounted. This method may be used for 

 connective tissue. The elastin stains dark brown ; the collagen, light brown. 

 Phosphotungstic acid hceniatoxylifi : 

 Haematoxylin, 1 gr. 



1 per cent, phosphotungstic acid (aqueous solution), 100 cc. 

 Heat the haematoxylin in a little water until it dissolves, and when cool add it to 

 the dilute acid. The solution is greenish-brown at first, changing to pale brown. 

 It may be used at once, keeps well, and requires no antiseptic. For elastic 

 fibers, striated muscle fibers, fibrin, and cartilage, the tissue is hardened in 

 alcohol, stained in the solution for twenty-four hours, washed in water, then 

 dehydrated and mounted in the usual way. Nuclei and elastic fibers are stained 

 blue ; connective tissue, pink. 



There are two neuroglia stains by Mallory — one with anilin-gentian violet, and 

 the other, which is here given, with the phosphotungstic haematoxylin solution. 

 a. Fix in 4 per cent, aqueous solution of formaldehyde for four days. 



