HEMATOIDIN 



154 HEMOCHROMOGEN CRYSTAL TEST 



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 difliculty 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 Iddney. 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 PorpJiyrins. 



Hematoxylin is the most useful of all dyes 

 in animal histology and pathology (Gr. 

 haimatodec, blood like + Xylon, wood). 

 It is an extract of logwood {Haernatoxy- 

 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 

 advantage. Ripening is brought about 

 by the formation of oxidation products. 

 Consequently it is recommended that 

 solutions be exposed to light and ajr. 

 Hematein (not hematin — a blood pig- 

 ment) is the oxidation product which 

 yields a fine deep blue coloration and is 

 the one most desired. It can be pur- 

 chased. To make up solutions of 

 hematein instead of hematoxylin is 

 logically sound but there is no way to 

 prevent further ripening (oxidation) 

 with the development of other browner 

 unwanted products and precipitation of 

 dyes. Therefore it is good practice to 

 begin with hematoxylin, to let it ripen 

 naturally over a fairly long period of time 

 or to ripen almost immediately by 

 adding about 5% hydrogen peroxide, or 



5% of 1% aq. potassium permanganate. 

 10% solution of hematoxylin in 

 96% or abs. ethyl alcohol should 

 always be kept on hand. It attains 

 maximum ripening in about one year, 

 but must be kept in a stoppered bottle 

 for otherwise the alcohol will evaporate. 

 It is diluted to 0.5% of hematoxylin 

 with aq. dest. for the Iron Hematoxylin 

 technique. See also Delafield's, Ehr- 

 lich's, Harris' and Mayer's hema- 

 toxylin solutions, likewise Azure 11 

 eosin and Hematoxylin. Addition of 

 a drop of Tergitol No. 7 to hematoxylin 

 solution will greatly increase speed of 

 staining but has no other advantage 

 (McClung 1950 p. 136). 



Hematoxylin and Eosin is rightly the most 

 used of all staining methods. If the 

 tissues have been fixed in a fluid con- 

 taining mercuric chloride such as Zen- 

 ker's fluid deparaffinize sections and 

 treat with dilute iodine in 70% alcohol 

 for 1-2 min. Wash in aq. dest., bleach 

 in 5-10% aq. sodium hyposulphite to 

 remove iodine and wash again in aq. dest. 

 Stain with Harris' Hematoxylin (full 

 strength) for 12-15 min. Blue in tap 

 water or in aq. dest. + few drops sat. 

 aq. lithium carbonate, 5-10 min. Stain 

 in 0.2% aq. eosin, 1 min. Rinse in aq. 

 dest. and 95% alcohol. Dehydrate in 

 absolute alcohol, clear in xylol and 

 mount in balsam. Nuclei, deep blue; 

 cytoplasm, pink. In place of Harris' 

 alum hematoxylin, which we use, 

 Delafield's Alum Hematoxylin or Ehr- 

 lich's Acid Hematoxylin may be em- 

 ployed. The Bensleys (p. 73) dilute 

 1 part of the last named with 2 parts 

 cold sat. aq. ammonium alum and 4 

 parts aq. dest. Nuclei, dark blue; 

 cytoplasm, collagenic fibers, erythro- 

 cytes, pink; smooth muscle, lavender. 

 0.2% aq. erj'throsin can take the place 

 of the eosin but the advantage is ques- 

 tionable. 



Hemin Crystal Test for blood pigment, 

 Teichmann (Stitt, p. 698). Dissolve in 

 100 cc. glacial acetic acid, 0.1 gm. of 

 KI, of K Br and of K CI. Add few 

 drops to suspected material on a slide 

 and cover. Gently warm until bubbles 

 begin, then slowly cool and examine for 

 typical dark brown crystals. The test 

 is not very sensitive but positive result 

 is conclusive. 



Hemochromatosis, clinical test for, see 

 Iron. 



Hemochromogen Crystal Test. Donogdny 

 (Stitt, p. 698). Mix 1 drop of suspected 

 fluid, of pyridin and of 20% aq. NAOH 

 on a slide and allow to dry. Radiating 

 crystals appearing within several hours 

 indicate presence of hemochromogen. 



