Journal of Applied Microscopy. 391 



Kleinenbet'g's Hvefnatoxyliji. — This stain has had a wide use, but is now 

 largely replaced by better formulae. It is mentioned here merely because it is 

 on the shelves of so many laboratories. It is doubtful whether it is equal to 

 Delafield's in any kind of botanical work. A good description is given in the 

 Quarterly Journal of the Microscopical Society, 74: 208 — , 1897. 

 Erlich^s Hematoxylin. — 



Distilled water, 50 cc. 

 Absolute alcohol, 50 cc. 

 Glycerine, 50 cc. 

 Glacial acetic acid, 5 cc. 

 Haematoxylin, 1 gr. 

 Alum in excess. 

 Keep it in a dark place until the color becomes a deep red. If well 

 stoppered, it will keep indefinitely. Transfer to the stain from 50 per cent, or 

 35 per cent, alcohol. Stain five to thirty minutes. Since there is no danger 

 from precipitates and the solution does not overstain, it is not necessary to treat 

 with water or with acid alcohol, but the slide may be transferred from the stain 

 to 70 per cent, alcohol. Eosin, erythrosin, or orange G are good constrast stains. 

 Boehmer's Hcematoxylin. — 



. ( Haematoxylin, 1 gr. . 



( Absolute alcohol, 12 cc. 



\ Alum, 1 gr. 



\ Distilled water, 240 cc. 



The solution A must ripen for two months. When wanted for use, add 

 about 10 drops of A to 10 cc. of B. Stain ten to twenty minutes. Wash in 

 water and proceed as usual. 



Mayer's H(zm-aliim. — Haematoxylin, 1 gr., dissolved with heat in 50 cc. of 

 95 per cent, alcohol and added to a solution of 50 gr. of alum in a liter of dis- 

 tilled water. Allow the mixture to cool and settle ; filter ; add a crystal of 

 thymol to preserve from mould {Lee). 



It is ready for use as soon as made up. Unless attacked by mould it keeps 

 indefinitely. Transfer to the stain from water. It is seldom necessary to stain 

 for more than ten minutes, and four or five minutes is generally long enough. 

 As a rule, better results are secured by diluting the stain (about 1 cc. to 10 cc. 

 of distilled water) and allowing it to act for ten hours or over night. This is a 

 good stain for the nuclei of filamentous algae and fungi, since it has little or no 

 effect upon cell walls or plastids. Wash thoroughly in water and transfer to 

 10 per cent, glycerine. (See Vol. 1, No. 9 of this Journal.) Specimens may be 

 mounted in balsam if they can be gotten through without shrinking. 



Haideiihairi' s Iron-Alum-HcBmatoxylin. — This stain was introduced by 

 Haidenhain in 1892 and has gained a well deserved popularity with those 

 engaged in cytological work. Two solutions are used and they are never mixed. 



A. \yn to 4 per cent, aqueous solution of ammonia sulphate of iron. (At 

 present we use a 3 per cent, solution.) 



B. One-half per cent, aqueous solution of haematoxylin. 



B 



