DS 23,213 DYE STAINS OF SPECIAL APPLICATION 477 



23.213 Gabbet 1887 test. 1928 Zinnser and Bayne-Jones 



Zinnser and Bayne-Jones 1928, 847 

 REAGENTS REQUIRED: A. DS 11.43 Zichl 1882; B. water 75, sulfuric acid 25, methylene 



blue 2 

 method: [heat-fixed smears] — > A, warmed to steaming, 2 mins. —* water, wash —* B, 1 

 min. — > water, wash —* dry 



23.213 Gibbe test. 1903 Cole Cross and Cole 1903, 160 



formula: water 45, 95% ale. 45, aniline 9, magenta 6, methyl blue 3 

 preparation: Grind dyes together and dissolve in aniline and ale. Dilute to 100 with 



water. 

 method: [heat-dried smears] -^ stain, heated to steaming, 4-5 mins. — * 95% ale, till no 



more color comes away -^ balsam, via clove oil 



23.213 Giinther 1898 Gunther 1898, 347 



reagents required: A. DS 11.43 Ziehl 1882; B. 3% nitric acid in 70% ale; C. sat, sol. 



(circ. 2%) methylene blue 

 method: [coverslip smear] — > A, in watch glass, heated to steaming, 1 min. —* B, 1 min. 



— » water, wash -^ C, on coverslip, few sees. —* water, wash —* dry 



23.213 Herman test. 1904 Besson Besson 1904, 638 



reagents required: A. 1% ammonium carbonate 80, 3% crystal violet 20; B. 10% 



nitric acid; C. 1% eosin in 60% ale. 

 method: [heat-fixed smear] —>■ A, heated to boiling, 1 min. — > B, 4-5 sees. — + abs. ale, 



till decolorized — > C, 30 sees. -^ abs. ale, quick rinse — > dry 



23.213 Koch 1884 1648, 2 :10 



reagents required: A. DS 23.211 Ehrlich 1882; B. 25% nitric acid; C. sat. sol. (circ. 

 1.5%) Bismarck brown 



method: [smears] —>■ A, 12 hrs. -^ B, momentary dip — > 60% ale, wash —y D, 15 sees. — > 

 water, wash -^ dry 



note: Much confusion exists in the literature about this technique, which is often at- 

 tributed to Ehrlich (loc. cit., A above) who recommended his solution for general, not 

 diagnostic, staining. The journal reference given above is technically incorrect for, in 

 the two years prior to 1886, the journal was known as the Mitteilung der Kaiserliche 

 Gesundheitsamte, which is, however, not recorded in the World List of Scientific 

 Periodicals. 



23.213 MuUer and Chermock 1945 11284, 30:169 



reagents required: A. DS 11.43 MuUer and Chermock 1945; 5. 3% hydrochloric acid 



in 95% ale, C. DS 11.44 MuUer and Chermock 1945 

 method: [heat-stained smears] -^ A, \ min. -^ B, 30 sees. — > C, 1 min. — » wash -^ dry 



23.213 Neelsen 1883 Ziehl-Neelsen — compl. script 23730, 21 :497 



reagents required: A. DS 11.43 Ziehl 1882; B. 25% nitric; C. DS 11.44 Loffler 1890 

 method: [sputum smears, heat-fixed] — > ^, on slide warmed to steaming, 3-5 mins. — ♦ 



water, rinse -^ B, till faint pink -^ wash -^ C, 2 mins. -^ wash — » dry 

 result: acid-fast bacteria, red; others, blue. 



note: The B solution is nowadays usually replaced by a 3% solution of hydrochloric acid 

 in 95% ale (Cowdry 1943, 17.) The term Ziehl-Neelsen has come to mean any acid- 

 differentiated magenta stain. Blot 1901 (6539, C. Lyons, 234) recommends 40% 

 formaldehyde in place of C above. A detailed description of the use of this technique 

 is given under DS 23.20 above. 



23,213 Pappenheim 1898 2813, 37 :809 



reagents required: A. DS 11.43 Ziehl 1882; B. abs. ale 100, glycerol 20, aurin 1, 



methylene blue to sat. (circ. 2) 

 method: [heat-fixed smear] —> A, heated to steaming, 2 mins. — > B, on slide, drained 

 and renewed 4-5 times -* water, wash —* dry 



