STAINING REAGENTS. 69 



a (loop blue, and other parts of some tissues light blue, e.g., the 

 matrix of hyaline cartilage. 



2. Strong Nucleus- Staining Hsematoxylin (Hamilton). 



Ha-matoxylin 

 Alum 

 Glycerine 

 Distilled water 



12 grams. 

 50 

 65 cc. 



130 



Boil, and while hot add 5 cc. liquid carbolic acid. Allow the 

 mixture to stand in the sunlight for at least a month. 



3. Delafield's Hsematoxylin. To 100 cc. of a saturated solution 

 of ammonia alum add drop by drop a solution of i gram hsematoxylin 

 dissolved in 6 cc. absolute alcohol. Expose to the air and light 

 for a week. Filter. Add 25 cc. glycerine and 25 cc. of methylic 

 alcohol. Allow it to stand exposed to the light for a long time. 

 Filter. 



This solution stains extremely rapidly, and may be greatly diluted 

 when it is used. It keeps for a very long time, and stains well 

 even tissues which have been hardened in chromic or osmic acid. 



4. Kleinenberg's Hsematoxylin. (i.) Make a saturated solution 

 of calcium chloride in 70 per cent, alcohol. Shake it well, and 

 allow it to stand. Decant the saturated solution, add alum to 

 excess, shake it well, allow it to stand for a day or so, and filter. 



(2.) Make a saturated solution of alum in 70 per cent, alcohol. 

 Filter. 



(3.) To one volume of the filtrate from (i) add eight volumes 



Of (2). 



(4.) To (3) add drop by drop a saturated solution of hsematoxylin 

 in absolute alcohol, until it becomes of a decided purple colour, 

 but not too dark, as the solution becomes darker by keeping and 

 exposure to light. It should be prepared at least a few months 

 before it is wanted. 



It may be diluted to any extent by adding the mixture (i) or 

 (2). As it contains much spirit, sections placed in it must be 

 covered, i.e., protected from evaporation, else the spirit will rapidly 

 evaporate. 



This logwood stain is particularly valuable when it is required 

 to stain a tissue or an organ " in bulk." This a diluted solution 

 will do in 24-48 hours, provided the pieces be not too large. 



5. Acid Hsematoxylin (Ehrlicli). 



Haematoxylih ..... i gram. 

 Absolute alcohol . . . 30 cc. 



To the solution add 



Glycerine . . . . . 60 cc. \ Saturated 



Distilled water . . . . . 60 ,, /with alum. 



Glacial acetic acid . . . 3 



