DS 23.215 DYE STAINS OF SPECIAL APPLICATION 483 



method: [heat-fixed smears, still warm] —* A, flooded on slide and warmed to steaming, 

 1 min. — » 90% ale. till color clouds cease — > B, flooded on slide and warmed to steam- 

 ing, 1 min. -+ water, rinse — > balsam, via usual reagents 



note: The original method recommended that either 1% sulfuric acid or 1% sodium 

 carbonate be added to A in amounts determined empirically for each organism . Zikes 

 1930 (23684, 81:161) substitutes his ADS 12.2 for A above. 



23.215 Loffler 1890b 23684, 7 :629 



REAGENTS REQUIRED: A. Water 80, tannin 20, ferric alum 5, sat. aq. sol. indigocarmine 

 10, sat. ale. sol. methyl violet 10; B. sat. aq. sol. aniline 100, crystal violet to sat, 10% 

 sodium carbonate 0.1 

 method: [heat-fixed smears] — > A, flooded on slide and warmed to steaming, 1 min. — ♦ 

 90% ale, till color clouds cease—* B, adjusted by trial and error to required pH, 

 flooded on slide and warmed to steaming, 1 min. — > wash -^ balsam, via usual 

 reagents 

 note: The formula for A above will come as a shock to those who have not consulted 

 the original reference. Most authors quote "ferrous sulphate" from the fact that the 

 solution of "ferrum sulphitricum oxydulatum ammoniatum" given on p. 629 (loc. cit) is 

 casually referred to as the f err osulph alios ung on p. 630. The formula for the B solution 

 comes from Loffler 1889 (23684, 6:213) and was there recommended for staining after 

 a hematoxylin mordant. 



23.215 Maneval 1929 20540b, 4:21 



REAGENTS REQUIRED: A. water 100, tannic acid 13, ferric chloride 1, DS 11.43 Ziehl 1882 



6.5, hydrogen peroxide (3%) 12; B. water 60, 95% ale. 27.5, aniline 2.5, acetic acid 



0.1, magenta 2 

 method: [smears of distilled water, suspensions of actively motile organisms] —* A, 2-4 



mins. — ♦ wash — » B, 2-3 mins. —> wash -^ dry 



23.215 Muir test. 1920 Stitt Stitt 1920, 56 



REAGENTS REQUIRED: .4. DS 23.217 Muir (1920) sol. B. 80, DS 11.43 Ziehl 1890 20; 



B. sat. sol. (circ. 4%) potassium alum 100, sat. ale. sol. (circ. 15%) methyl violet 20 

 method: [heat-fixed smears] -^ A, steamed over bath, 1 min. -^ water, 2 mins. — + dry —* 



B, on slide, warmed to steaming, 1 min. -^ water, wash -^ dry 



note: Both A and B solutions are unstable. 



23.215 Nicolle and Morax test. 1904 Besson Besson 1904, 169 



REAGENTS REQUIRED: A. DS 23.215 Loffler 1890b, (sol. A); B. DS 11.43 Ziehl 1882 

 method: [heat-fixed smear] -^ A, on slide, heated to steaming, 3 or 4 changes —> B, on 

 slide, heated to steaming, 3 or 4 sees. — ♦ wash -^ dry 



23.215 Novel 1939 see MS 33.52 Novel 1939 



23.215 Pittfield test. 1910 Heymann Ehrlich, Krause, et al. 1910, 2, 394 



STOCK solutions: I. 10% aluminum acetate 10, sat. ale. sol. crystal violet 100, II. 10% 



tannic acid 

 WORKING solution: stock I 50, stock II 50 



method: [heat-fixed smears] — > stain, lightly warmed -^30% ale. till differentiated -* dry 

 notes: Wright 1928 (20540b, 3 :17) cites (with reference to Kendall 1902: 11032, 5 :1936) 

 a method of Pittfield 1902 in which the mordant contains "sat. sol. alum." 



23.215 Remy and Sugg test. 1904 Besson Besson 1904, 169 



REAGENTS REQUIRED: A. DS 23.215 Loflfler 1890 (sol. /I); B. ADS 12.2 Gram 1884; C. sat. 



aq. sol. aniline 80, sat. ale. sol. {circ. 10%) crystal violet 0.2, water 20 

 method: [heat-fixed smear] -^ A, 15-30 mins. -^ B, on slide, few moments -^ wash — > 



abs. ale, wash — > C, 30 mins. at 37°C. -^ wash — > balsam, via usual reagents 



23.215 Rossi test. 1904 Besson Besson 1904, 170 



RE.\GENTs required: .4. ADS 12.2 Rossi (1904); B. DS 11.43 Ziehl 1S82 

 method: [heat-fixed smear] -^^ ^4, 30 mins. — ♦ wash — » dry —* B, heated to steaming, 3-4 

 mins. — > wash — > dry 



