HEMATOXYLIN 



118 



HEMOPHILUS PERTUSSIS 



advantage. Ripening is brought about 

 by the formation of oxidation products. 

 Consequently it is recommended that 

 solutions be exposed to light and air. 

 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. eth)^ alcohol should 

 alwaj^s 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 hematoxjdin 

 with aq. dest. for the Iron Hematoxylin 

 technique. See also Delafield's, Ehr- 

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

 toxylin solutions, likewise Azure II 

 eosin and Hematoxylin. 



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 inaq. dest. 

 Stain with Harris' Hematoxylin (full 

 strength) for 12-15 min. Blue in tap 

 water or in aq. dest. -f 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 maj' be em- 

 ployed. The Benslej'-s (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. erythrosin can take the place 

 of the eosin but the advantage is ques- 

 tionable. 



Kemin 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, Donogany 

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

 fluid, of pyridin and of 20% aq. NAOH 

 on a slide and allow to dr}'. Radiating 

 crystals appearing within several hours 

 indicate presence of hemochromogen. 



Hemocytoblasts, see Erythrocytes, develop- 

 mental series. 



Hemofuscin. Mallory's fuchsin stain. Fix 

 in Zenker's fluid, alcohol or 10% forma- 

 lin. Stain nuclei in paraflin or celloidin 

 sections with Iron Hematoxylin. Wash 

 thoroughly in water. Stain 5-20 min. 

 in : basic fuchsin 0.5 gm., 95% ale. 50 cc. 

 and aq. dest. 50 cc. Wash in water. 

 Differentiate in 95% alcohol, dehydrate 

 in abs. ale, clear in xylol and mount in 

 balsam in the case of paraffin sections. 

 Celloidin sections are to be cleared in 

 terpineol or origanum oil after 95% ale. 

 Nuclei blue, hemofuscin granules bright 

 red, hemttsiderin and melanin unstained 

 (Mallory, p. 136). 



Hemoglobin, histochemical test (Ralph, 

 P. IL, Stain Techn., 1941, 16, 105-106). 

 Flood dried blood smear with 1% 

 benzidine in absolute methyl ale, 1 

 min. Pour off and replace with 25% 

 superoxol in 70% ethyl ale, 90 sec. 

 Wash in aq. dest., 15 sec. Dry and 

 mount in neutral balsam. Hemoglobin 

 dark brown. 



Hemoglobin Estimation is done by compar- 

 ing blood with a colored paper scale or bj' 

 a more accurate scale in a hemoglobinom- 

 eter. The experimental error is at 

 least 5%. Staining reactions for hemo- 

 globin within cytoplasm (Kindred, J. 

 E., Stain Techn., 1935, 10, 7-20). 



Hemolysis. Methods for measuring the 

 velocity of hemolysis depend on the 

 fact that red blood cell suspensions as 

 ihey hemolyse become more and more 

 translucent. Techniques differ merely 

 in the v/aj's of measuring the trans- 

 mitted light. Simple visual photom- 

 eters and photoelectric ones are de- 

 scribed by Ponder, E. Glasser's Medical 

 Ph5'-sics, 605-612. The same authority 

 explains the "equilibrium methods" for 

 measuring the amount of hemolysis 

 which has taken place if the process has 

 been arrested. One of these is to count 

 the cells remaining, another to deter- 

 mine the amount of hemoglobin set 

 free, etc. 



Hemophilus Pertussis. Staining of cap- 

 sules in air dried smears with 5% aq. 



