AUER BODIES 



AUER BODIES 



1. Unstained moisl preparations: A 

 drop of blood or bone marrow is placed 

 on a clean cover slip, inverted on a slide 

 and rimmed with vaseline or suitable 

 substance to prevent drying. The prepa- 

 ration is examined using the phase 

 microscope. Auer bodies appear as dis- 

 crete cytoplasmic structures. 



2. Supravital technic: The supravital 

 technic is carried out in the same man- 

 ner as indicated above (No. 1.) for un- 

 stained moist preparations with the 

 exception of the fact that the slide has 

 been previously coated with neutral 

 red and janus green. Preparations may 

 be examined with the bright field, dark 

 field or phase microscope. Under the 

 bright field microscope, the Auer bodies 

 appear as discrete structures which 

 stain with neutral red. The color of the 

 Auer bodies varies from a deep red 

 (acidic reaction) in young cells to tan 

 (less acidic reaction) in the more mature 

 leukemic cells. The staining reaction 

 of the Auer bodies with neutral red 

 closely parallels that of cytoplasmic 

 granules. Auer bodies stain blue-purple 

 in supravital preparations prepared 

 with brilliant cresyl blue (Dameshek, 

 W. Arch. Int. Med., 1932, 50, 579). 



3. Romanowsky stains: The Roman - 

 owsky dyes may be used to demonstrate 

 Auer bodies. Due to the solubility of the 

 azurophilic component of the Auer 

 body in water and alcohol, this technic 

 is not as satisfactory as the supravital 

 and the unstained moist technics. 



4. Swlan black B technic: Dry blood 

 and bone marrow films are fixed with 

 either formalin vapor or with a 1% solu- 

 tion of formalin in 95% ethyl alcohol 

 for 20-60 seconds. The dried-fi.xed films 

 are placed in a saturated solution of 

 Sudan black B (National Aniline Co., 

 New York) in 70% ethyl alcohol for 

 20-30 minutes. The solution of Sudan 

 black must be prepared at least 2 weeks 

 prior to using. The blood films are then 

 dipped in 70% ethyl alcohol and rinsed 

 in 50% ethyl alcohol and washed in dis- 

 tilled water, dried and mounted with 

 "permount." Auer bodies are sudano- 

 philic and appear black or black-brown. 

 This technic is an excellent method for 

 demonstrating Auer bodies. 



5. Periodic Acid-Schiff {PAS) re- 

 action: Dried films are fi.xed with forma- 

 lin vapor for 20-60 seconds. The fixed 

 preparations are immersed in an 

 aqueous 0.5% solution of periodic acid 

 for 5 minutes, washed thoroughly with 

 distilled water and immersed in Schiff's 

 reagent 10 minutes, followed by 3 suc- 

 cessive changes in sulfurous acid, 3-5 

 minutes each, washed in distilled water, 

 dried and mounted with "permount." 



Auer bodies are PAS-positive appear- 

 ing a moderate pink. 



6. Toluidine blue ami Thionin: Dried 

 films may be fixed with formalin vapor, 

 4% aqueous basic lead acetate or other 

 suitable fixatives. Preparations are 

 washed, dried and immersed in a dilute 

 solution (0.2-0.5%) toluidine blue or 

 thionin, washed in distilled water, dried 

 and mounted with "permount." Auer 

 bodies stain lavender following the use 

 of this technic. 



7. Plasinal reaction: Dried films are 

 fixed with formalin vapor for 20-60 

 seconds, washed in distilled water and 

 placed in a 1% aqueous solution of 

 mercuric chloride for 5 minutes, washed 

 in distilled water and placed in Schiff's 

 reagent; the films are placed in three 

 successive changes of sulfurous acid, 

 washed in distilled water, dried and 

 mounted with "permount." In staining 

 control preparations, the same pro- 

 cedure is followed with the exception 

 of the fact that distilled water is sub- 

 stituted for mercuric chloride. If the 

 control exhibits a positive reaction, the 

 length of exposure to Schiff's reagent 

 must be reduced. Following the plasmal 

 reaction, the Auer bodies appear a very 

 pale pink, and may best be identified by 

 means of the phase microscope followed 

 by bright field observation of the 

 stained bodies. 



8. M. nmli reaction: Dried blood and 

 bone marrow films may be fi.xed either 

 with formalin vapor or 1% formalin in 

 95% ethj'l alcohol, washed with dis- 

 tilled water and dried. The fi.xed films 

 are immersed in a mixture containing 

 equal parts of 1% alkaline solution of 

 alpha-napthol and 1% aqueous solution 

 of p-aminodimeth_ylanaline (Eastman 

 Kodak, P2147) for 1-5 minutes, washed 

 in distilled water and mounted in water. 

 Auer bodies stain deep blue following 

 the M. nadi reaction. Preparations fade 

 rapidly. 



9. Sato and, Sekeya Peroxidase Technic: 

 Dried blood and bone marrow films are 

 placed in a 0.5% aqueous solution of 

 copper sulfate for 30 seconds, washed 

 briefly in 50% ethyl alcohol and placed 

 in a benzidine-hydrogen peroxide mix- 

 ture (0.1 gm. benzidine in 100 ml. dis- 

 tilled water and 2 drops of 3% hydrogen 

 peroxide) for 2 minutes, washed with 

 distilled water, dried and mounted with 

 "permount". Counterstaining with 1% 

 aqueous safranin is optional. Auer 

 bodies are peroxidase-positive and ap- 

 pear blue or j'ellow. 



Auer bodies are soluble in water, 

 saline and many organic solvents and 

 are readily digested bj' ribonuclease. 

 Digestion by ribonuclease may be pre- 



