486 METHODS AND FORMULAS DS 23.216 



method: [heat-fixed smears] —>■ A, heated to bubbling -» cool -^ [repeat twice] -* wash 

 -^ blot -* B, warmed to steaming -^ cool -^ repeat twice -* C, till decolorized -* 

 wash -> D, 30 sees. —>■ wash -^ dry 



23.216 Gray 1941 14900, 147:329 



REAGENTS REQUIRED: A. Water 100 malachite green 0.5, magenta 0.05 

 method: [heat-fixed smears] -^ A, heated to steaming, 1 min. — * wash —>■ dry 



23.216 Kahlden and Laurent 1896 Kahlden and Laurent 1896, 101 



REAGENTS REQUIRED: A. DS 11.43 Ziehl 1882; B. 5% nitric acid; C. DS 11.44 Loffler 1890 

 method: [heat-fixed smear] -^ A, 20-40 mins. -^ B, 1 min. -> wash -^ C, 2 mins. -> 

 wash —y blot — » dry 



23.216 Lagerberg 1917 23684, 79:191 



REAGENTS REQUIRED: A. sat. sol. {circ. 25%) copper sulfate; B. 20% ammonium hy- 

 droxide; C. DS 11.43 Ziehl 1882; D. 4% sulfuric acid 

 method: [heat-fixed smears]-^ A, flooded on slide -^ B, added drop by drop till ppt. 

 just redissolved -^ warm to steaming — » add several drops A-* B, rinse -^ water, 

 wash -> C, on slide, heated to steaming, 1 min. -^ D, till no more color comes away 

 — > water, wash —* dry 



23.216 May 1926 20540b, 1:105 



REAGENTS REQUIRED: A. 5% chromic acid; B. ammonia; C. DS 11.43 Ziehl 1882; D. 1% 



sulfuric acid; E. DS 11.44 Loffler 1890 

 method: [heat-fixed smear] -> A, on slide, 30 sees. -^ B, added (twice as much) to A 



on slide; 2 min. -^ rinse -* C, on slide, heated to steaming, 2-3 mins. -^ rinse -^ D, 



15-30 sees. -> wash — > E, few drops added to water on slide, 10-30 sees. — » rinse -»■ 



blot — > dry 



23.216 MoUer 1891 23684, 10:9 



REAGENTS REQUIRED: A. chloroform ; B. 5% chromic acid; C. DS 11.43 Ziehl 1882; D.1% 



sulfuric acid; E. 1% methylene blue 

 method: [heat-fixed smears] -^ A, 1-2 mins. -^ abs. ale, rinse -^ water, wash ^- B, 1 

 min. -^ water, wash -^ B, 1 min. -^ water, wash — > C, on slide, heated to steaming, 

 5 mins. -♦ water, wash -^ D, till faint pink -^ water, wash —> E, 10 sees. -^ water, 

 wash — » dry 



23.216 Muzzarelli 1931 test. 1942 Langeron Langeron 1942, 1216 



reagents required: A. water 75, DS 11.44 Manson 25; B. 10% nitric acid; C. 1% eosin 



method: [heat-fixed smears]—* A, warmed to steaming, 30 sees. — » water, wash -^ B, 



till no more color comes away — > water, wash — > C, 15 sees. — > water, wash — > dry 



23.216 Neisser and Hueppe test. 1928 Schmorl Schmorl 1928, 355 



reagents required: A. sat. sol. magenta in sat. sol. aniline; B. 25% sulfuric acid; C. 



1 % methylene blue 

 method: [heat-fixed smear] -^ A, 100°C., 1-5 hrs. — > B, 5 sees. -^ abs. ale, till no more 



color comes away — > water — + C, 3-5 mins. — > wash -^ dry 

 note: Schmorl {loc. cit.) recommends a steam chest for step A. 



23.216 Proca 1909 6630,68:307 



formula: water 50, DS 11.44 Loffler 1890 50, DS 11.43 Ziehl 1882 4 

 method: [heat-fixed smears] — > stain, 1 min. -^ water, wash -^ dry 

 result: live bacteria, colorless spores on blue; dead bacteria, blue spores on red. 

 note: See also DS 23.21 Gay and Clark 1924, who have adapted this technique to 

 general use. 



23.216 Ruiz 1946 test. 1947 Pemander 20540b, 22:164 



reagents required: A.1% methyl violet; B.0.1% eosin Y 100, normal horse serum 30 

 method: Mix 1 drop A with bacterial suspension, leave 5 mins. Add 1 drop B, leave 3 

 mins. Make smear and dry. 



