154 HMMATEIN (HMMATOXYLIN) STAINS. 



KAWITZ (Leitfaden, 2nd ed., p. 63) takes 1 grin, haematein, 6 grms. 

 ammonia alum, 200 grms. each of water and glycerin. 



Or (Zeit. wiss. Mik., xxv, 1909, p. 391) 1 grm. hsematein, 10 grms. 

 of nitrate of aluminium, 250 grms. each of water and glycerin. 



252. HANSEN'S Solution (Zool. Anz. t 1895, p. 158). See fourth edition. 



253. HARRIS'S Solution (Micr. Bull, xy, 1898, p. 47 ; Journ. App. 

 Mic., iii, p. 777). Alum-hsematoxylin solution ripened by addition of 

 mercuric oxide. MAYER (Grundzuge, 1901, p. 171) finds the formula 

 " gives too much hsematein." 



254. Bohmer's Heematoxylin (Arch. mik. Anat., iv, 1868, p. 345 ; 

 Aerzt. Intelligenzbl., Baiern., 1865, p. 382). Make (A) a solution of 

 haematox. cryst. 1 part, alcohol (absolute) 12 parts, and (B) alum 

 1 part, water 240. For staining, add two or three drops of A to a 

 watch-glassful of B. 



The alcoholic solution of hsematoxylin ought to be old and dark 

 ( 237). 



A. Gr. HORNYOLD (Trans. Manch. Micr. Soc., 1915) prepares : 

 Solution A : haematoxylin, 0-7 grm. ; absolute alcohol, 20 c.c. Solution 

 B : alum, 0-35 grm. ; aq. dest., 60 c.c. Mix A and B, expose to light 

 in window for three or four days, then add 20 drops of tincture of 

 iodine. Stain sections five to ten minutes till red-brown. Differentiate 

 in 70 per cent, alcohol, to which add a few drops of acetic acid. The 

 sections then turn blue. Grood stain after osmic fixatives. 



255. Delafield's Hsematoxylin (Zeit. wiss. Mik., ii, 1885, p. 288 ; 

 frequently attributed erroneously to GRENACHER or PRUDDEN). 

 To 400 c.c. of saturated solution of ammonia-alum (that is about 

 1 to 11 of water) add 4 grms. of hsematox. cryst. dissolved in 25 c.c. 

 of strong alcohol. Leave it exposed to the light and air in an un- 

 stoppered bottle for three or four days. Filter, and add 100 c.c. of 

 glycerin and 100 c.c. of methylic alcohol (CH 4 0). Allow the solution 

 to stand (uncorked) until the colour is sufficiently dark, then filter. 



This solution keeps for years. It is well to allow it to ripen for 

 at least two months before using it. 



For staining, enough of the solution should be added to pure water 

 to make a very dilute stain. It is an extremely powerful stain. 



It is still much used. I find that When well ripened for years 

 rather than months it is quite a first-class stain. 



BUTSCHLI (Unters. ub. mikroscopische Schdume u. das Protoplasma, 

 etc., 1892) recommends, under the name of "acid haematoxylin," 

 solution of Delafield very strongly diluted, and with enough acetic acid 

 added to it to give it a decidedly red tint. This gives a sharper and 

 more differentiated nuclear stain than the usual solution. 



MARTINOTTI (Zeit. wiss. Mik., xxvii, 1910, p. 31) makes it up with 0*2 

 per cent, of hcematein, and less alum (2 per cent.). 



