494 METHODS AND FORMULAS DS 23.222 



23.222 Brown and Brenn 1931b 11056, 21:21 



STOCK solutions: I. water 100, gentian violet 1; II. water 100, sodium bicarbonate 5, 



phenol 0.5 

 REAGENTS REQUIRED: A. stock I 100, stock II 11; 5. ADS 12.2 Lugol (1905); C. acetone 



75, ether 25; D. 0.005% magenta; E. 0.1% picric acid in acetone 

 method: [sections, nuclei stained by DS 11.122 technique]—* waters A, on slide, 2 

 mins. — > rinse — > 5, 1 min. — > wash — > blot — » C, on slide, till no more color comes 

 away -^ blot D, 5 mins. — > wash — » blot E, till color turns to yellowish pink -^ bal- 

 sam, via acetone and xylene 



23.222 Glynn 1935 1789a, 20 :896 



REAGENTS REQUIRED: A. DS 11.45 Crystal violet; B. ADS 12.2 Gram (1884); C. acetone; 



D. water 100, magenta 0.05, hydrochloric acid q.s. for pH 2.5; E. sat. sol. picric 



acid 

 method: [sections] —* water -^ A, 2 mins. — > drain -^ B, on slide, 1 min. — > C, till no 



more color comes away — > water, wash — > Z), 3 mins. —>■ drain -^ E, on slide, 1 min. 



-^ C, 10-15 sees. —> balsam, via xylene 

 result: Gram-positive bacteria, violet; Gram-negative bacteria, red. 



23.222 "Gram-Weigert" — compl. script. 



The original method is Weigert 1887 (see below). Almost any technique involving the 

 differentiation of gentian violet with iodine followed by aniline has come to bear this 

 name. Unfortunately Weigert's 1898 (23681, 9:290) resorciii-magenta method (see 

 DS 21.13 Weigert 1898) has also become confused with the 1887 method. Authors who 

 state only that they "stained by the Gram-Weigert method" may have used crystal 

 violet or magenta, with or without prior nuclear staining in carmine (Zinsser, Bayne- 

 Jones, 1939, 860) or hematoxylin (Mallory, 1938, 272), with or without aniline 

 differentiation. 



23.222 Haythorn 1929 4349, 12:128 



REAGENTS REQUIRED: A. DS 11.122 Mallory 1938; B. 0.1% hydrochloric acid in 70% 

 ale; C. DS 23.211 Ehrlich 1882; D. ADS 12.2 Lugol (1904); E. aniline 60, xylene 30; 

 F. sat. ale. sol. erythrosin; G. aniline 30, xylene 60 

 method: [sections] — + water —>■ A, 5-10 mins., or till deeply stained — > B, quick rinse — > 

 tap water, wash — > C, 2-5 mins. — > blot -^ D, 2-5 mins. — * E, till differentiated — » F, 

 30-60 sees. -^ G, till no more color comes away — > balsam, via xylene 



23.222 Krajian 1943 11284,28:1602 



reagents required: A. DS 11.44 Loffler 1890; B. xylene 65, creosote 35; C. DS 11.43 



Krajian 1943 

 method: [sections]—* water—* A, 3 mins. -^' wash—* abs. ale, least possible time for 



dehydration -^ B, till differentiated, 2-5 sees. — » C, on slide, 2 changes, 10 sees, each 



—> blot — * B, on slide till differentiated — > blot — * balsam, via xylene 

 result: nuclei. Gram-negative organisms, Negri bodies, red; Gram-positive organisms, 



actinomycetes, blue. 



23.222 Krajian 1950 Krajian 1950, 196 



REAGENTS REQUIRED: A. 5% thorium nitrate; B. DS 11.121 Krajian 1950; C. 1% hydro- 

 chloric acid in 70% ale, D. water 100, potassium iodide 5, zinc sulfate 5; E. DS 11.43 

 Ziehl 1882; F. 2% sodium sulfite; G. 3% acetic acid; H. 50:50 aniline xylene; I. cresote 

 method: [5 ju to 7 m sections] — > water — * A, 5 mins. -^ rinse -^ B, 3 mins. -^ wash — * C, 

 till no more color comes away — * tap water, till blue -^ D, 5 mins. — > rinse — * E, 7-10 

 mins. — > wash — * F, 2 mins. -^ rinse — * G, 3 mins. -^ blot — * H, 30 sees. — * /, till 

 pink clouds cease -+ dammar, via xylene 

 result: Gram-positive organisms, blue black; Gram-negative organisms, red. 



23.222 Male 1924 11035,42:455 



reagents required: A. 0.5% methyl violet; B. ADS 12.2 Gram 1880; C. water 75, 



95% ale. 25, light green 0.05, neutral red 0.25 

 method: [sections] -^ water -^ A, 1-2 mins. -^ B, 1-2 mins. -* 95%, ale, till no more 



color comes away -^ C, 3-5 mins. -^ wash -> balsam, via usual reagents 



