HEIDENHAIN'S AZAN STAIN 



117 



HEMATOXYLIN 



Heidenhain's Azan Stain (Heidenhain, M., 

 Ztschr. f . wiss. Mikr., 1915, 32, 361-372). 

 The following details are from Lee (1928, 

 p. 279): Color sections 1 hr. at 55°C. 

 in 2% aq. azocarmine plus 10 drops 

 p;lacial acetic acid in small staining jar. 

 Wash in water. Differentiate in 96% 

 ale. 100 cc. plus anilin oil 0.1 cc. until 

 cytoplasm becomes pale pink and nuclei 

 clear red. To hurry differentiation add 

 2 drops anilin oil. Rinse in 96% ale. 

 containing few drops acetic. Put in 

 5% aq. phosphotungstic acid about 2 

 hrs. until connective tissue is com- 

 pletely decolorized. Wash rapidly in 

 water. Stain |-3 hrs. in following solu- 

 tion diluted with equal or double parts 

 aq. dest. : anilin blue (water sol. Griib- 

 ler) 0.5 gm.; orange G, 2 gm.; acetic 

 acid, 8 cc; aq. dest. 100 cc. Examine 

 staining under microscope. Wash in 

 water, dehydrate in abs. ale, clear in 

 xylol and mount in balsam. This is a 

 very useful stain. See also McGregor, 

 L., A.m. J. Path., 1929, 5, 545-557 for use 

 of this technique particularly as applied 

 to normal renal glomerules. Under 

 Islets of Langerhans is given use of a 

 slightly modified azan method by 

 Gomori. 



Heidenhain's Iron Hematoxylin, see Iron 

 Hematoxylin. 



Heinz Bodies. These spherical bodies are 

 sometimes seen in erythrocytes espe- 

 cially when examined in the dark field or 

 when colored with Azur 1. They have 

 been referred to as Substantia Meta- 

 chromatica Granularis and B-substance. 

 The best way to demonstrate them is 

 to use the technique of Figge, F. H. J., 

 Anat. Rec, 1946, 94, 17. Give 0.3% aq. 

 sulfanilamide to mice as drinking water. 

 Within 4-6 days these bodies will appear 

 in at least 90% of erythrocytes whence 

 they are cast out into the plasma. 

 They are most readily seen in unstained, 

 unmounted blood smears. They dis- 

 appear when studied in oil, balsam or 

 other mounting media. Heinz bodies 

 are granules of heme-containing pro- 

 tein denatured by this drug within the 

 cells. They are not produced by sod- 

 ium sulfathiazole. 



Helianthin, see Methyl Orange, 



Heliotrope B, see Amethyst Violet. 



Helium, see Atomic Weights. 



Helly's Fluid is Zenker's fluid in which 5% 

 formalin is substituted for 5% acetic 

 acid. 



Helminthosporia. Stain for nuclei in (Par- 

 ris, G. K., Phvtopathology, 1944, 34, 

 700). 



Hemalum (Mayer's) Hematin, 1 gm.; 90% 

 ale, 50 cc. ; aq. dest., 1000 cc. ; ammonia 

 alum, 50 gms.; thymol, 1 crystal. 

 Keeps better after adding 20 cc. glacial 



acetic acid and making Acid Hemalum. 



A good nuclear stain when diluted with 

 aq. dest. 1:20. The above formula has 

 been modified by Lillie (R. D., Stain 

 Techn., 1942, 17, 89-90): hematoxylin, 

 5 gm.; sodium iodate (NalOj), 1 gm.; 

 ammonia alum (AINH4 (804)2 + 12 

 H2O), 50gm.;aq. dest., 700 cc, glycerol, 

 300 cc, glacial acetic acid, 20 cc. No 

 ripening is necessary. Stain sections 

 formalin fi.xed material, 2-5 min. Blue 

 2-10 min. in tap water. Counterstain 

 in 0.2% aq. eosin Y. Dehydrate clear 

 and mount as usual. This method is 

 quick and gives a sharp stain. 



Hematin, identified by luminescence with 

 Luminol. Do not confuse with hema- 

 tein, see Hematoxylin. 



Hematocrit, a tube used to concentrate red 

 blood cells by centrifugation and to 

 measure their volume, see Ponder, E. 

 in Glasser's Medical Physics, 597-600. 



Hematoidin (hematin + G. eidos, appear- 

 ance). An iron free pigment produced 

 by phagocytic digestion of erythrocytes 

 or in clots and old hemorrhages, chemi- 

 cal composition similar or identical with 

 Bilirubin. Seen as red or orange rhombic 

 plates or radiating yellow needles, 

 insoluble in ether, water and soluble only 

 with difficulty in alcohol, easily soluble 

 in chloroform. Gives positive Gmelin's 

 test. 



Hematoporphyrin (G. haima, blood + 

 porphyra, purple). — Written by Frank 

 H. J. Figge, Dept. of Anatomy Univer- 

 sity of Maryland Medical School, 

 Baltimore, Md. Contrary to a deeply 

 rooted misconception, this substance 

 is not the pigment as it occurs in hemo- 

 globin, but is artificially produced by 

 the drastic decomposition of hemo- 

 globin in concentrated strong acids. 

 Since it does not occur in nature, such 

 terms as "hematoporphyrinuria" are 

 obsolete. In addition, protoporphyrin, 

 which is the true, unaltered, pigment 

 found in heme compounds, is not ex- 

 creted as such by the kidney. Proto- 

 porphyrin is heme minus iron and has 

 two vinyl group side-chains. Hemato- 

 porphyrin is heme minus iron, plus two 

 hydrogen and two hydroxy! groups. 

 Hematoporphyrin is soluble in water, 

 ether, alcohols, dilute alkalies, and acids. 

 For references and additional informa- 

 tion, see Porphyrins. 



Hematoxylin is the most useful of all dyea 

 in animal histology and pathology (Gr. 

 haimatodec, blood like -f- Xylon, wood). 

 It is an extract of logwood (Haematoxy- 

 lon campechianum) and is marketed in 

 crystalline form. When the crystals 

 are first dissolved in water or alcohol it 

 is not an energetic stain; but requires 

 to be "ripened" before it can be used to 



