496 METHODS AND FORMULAS DS 23.223 



23.223 Bertrand and Medakovitch 1924 test. Langeron 1942 



Langeron 1942, 1204 

 REAGENTS REQUIRED: A. sat. sol. aniline 90, 95% ale. 10 magenta 5; B. 3% hydrochloric 



acid; C 1% lithium carbonate; D. 1% malachite green 

 method: [paraffin sections of F 1500.0010 fixed material] -^^ water -^ A, 1 min. 60°C. 

 -^ water, rinse -^ B, till no more color comes away — > water, wash -^ C, 1 min. -^ D, 

 30 sees. — > balsam, via usual reagents 

 RECOMMENDED FOR: acid-fast bacilli in sections. 



23.223 Campbell 1929 4349, 12:129 



REAGENTS REQUIRED: A. DS 11.43 Kinyoun (1946); B. 0.5% hydrochloric acid in 35% 



ale; C. DS 11.122 "Harris"; D. 1% ammonia; E. 1% orange G 

 method: [paraffin sections of F 3700.0010 Zenker 1894 fixed material]^ water -> A, 

 30 mins. -^ rinse -^ B, 2 changes, till pink -^ C, 2 mins. -^ B, till nuclei differenti- 

 ated -^^ D, till nuclei blue — > wash ^^ E, till plasma orange—* balsam, via acetone 

 and xylene 

 recommended for: demonstration of B. leprae (red) in sections. Myelin sheaths also 

 stain red. 



23.223 Czaplewski test. 1896 Kahlden and Laurent Kahlden and Laurent 1896, 117 



reagents required: .4. DS 11.43 Ziehl 1882; B. 95% ale. 100, fluorescin to sat., 



methyl blue to sat. ; C. sat. ale. sol. methyl blue 

 method: [sections] —> water ^ a, 3-5 mins., 37°C. ^ drain -+ B, 12 dips, draining 



slowly between each — > C, flooded on slide, 1 min. -^ rinse — > dry -^ balsam 



23.223 Doubrow 1929 4285a, 6:142 



REAGENTS REQUIRED: A. DS 11.43 Ziehl 1882; B. 4.5% picric acid in 95% ale; C. 0.5% 



light green in 95% ale. 

 method: [sections, nuclei stained by DS 11.111 Regaud 1910] -> A, 45 mins., 50°C. 



— > B, till only nuclei appear stained -^ water, till yellow stain removed — > C, 1 min. 



— > balsam, via usual reagents 



23.223 Douglas 1932 11284, 17:1131 



REAGENTS REQUIRED: A. DS 11.43 Ziehl 1882 100, sodium chloride 0.3; B. 5% nitric 



acid in 95% ale.; C. 1% azur II 5, DS 11.44 Jadassohn (1928) 5, water 90; D. ADS 



22.1 Wolbach 1911 

 method: [paraffin sections] ^ A, overnight -^ refrigerator, 30 mins. -^ wash —> 5, 1 



min. — * wash -^ C, 10 mins. — > wash -^ D, till differentiated -^ balsam, via usual 



reagents 



23.223 Fits 1939 11284,25:743 



REAGENTS REQUIRED: A. water 90, methanol 10, phenol 5, new magenta 0.5; B. 20% 



formaldehyde; C. 2% hydrochloric acid in 95% ale; D. 1% potassium permanganate; 



E. 2% oxalic acid; F. DS 11.123 Harris 1900; G. DS 12.221 Fite 1939 

 method: [sections of formaldehyde-alcohol-fixed material] — * water —> A, 12-24 hrs. — + 



B, 5 mins. — > C, 5 mins. — > rinse -^ D, 2-5 mins. —^E,! min. wash — > F, 2 mins. 



— » wash —* G, till stained — > 95% ale, — * balsam, via usual reagents 



23.223 Flexner test. 1938 Mallory Mallory 1938, 276 



REAGENTS REQUIRED: A. DS 11.43 Ziehl 1882; B. ADS 12.2 Gram (1904); C. aniline 

 method: [paraffin sections, nuclei stained by any DS 11.122 technique] —> water —> A, 



1 hr. — * water, wash -^ B, ^2 to 1 min. -* water, wash -^ blot — * C, till differentiated 



—*■ balsam, via xylene 



23.223 Fuller 1938 11284,23:416 



reagents required: A. 5% ferric alum; B. water 80, glycerol 10, 95% ale. 10, hema- 



toxyUn 1; C. 5% picric acid in 95% ale; D. 1.5% magenta in sat. aq. sol. aniline; E. 



3% nitric acid in 95% ale; F. 0.01% ammonia; G. 1% light green 

 method: [sections] — > water—* A, 5 mins., 50°C. -^ rinse-* B, 5 mins. 50°C. -* C, till 



nuclei only stained — > D, on slide, heated to steaming 3 mins. — > cool — » reheat, 3 



mins. —> E, till sections pale pink —* F, few sees. — > wash, 10 mins. — > G, 5 mins. — > 



rinse — > balsam, via usual reagents 

 result: bacilli, red; nuclei, blue green; cytoplasm, pale green. 



