352 METHODS AND FORMULAS DS 13.21-DS 13.22 



method: [water] ^ stain, 24 hrs. — > abs. ale, minimum possible time —> balsam, via 



xylene 

 note: Houcke {loc. cit.) recommends varying 'the acidity of the working solution by 



experiment to adapt it to use after various fixatives. 



13.21 Holmes and French 1926 azur C-eosin Y-orange II 



20540b, 1:25 

 REAGENTS REQUIRED: A. 1.5% azur C; B. abs. ale. 99, acetic acid 1, eosin Y 0.025, 



orange II 0.025 

 method: [water] -^ A, 5 mins. — > methanol, till color clouds cease, 5-10 sees. -^ B, till 



no more blue comes away -^ abs. ale, quick rinse — * balsam, via xylene 

 result: nuclei, and some bacteria, blue; cell inclusions and blood, as Giemsa; collagens, 

 bright orange; muscle, pink. 



13.21 Kedrovsky 1931 see DS 22.12 Kedrovsky 1931 



13.21 Mann 1894 test. 1942 Langeron cit. Masson erythrosin-orange G-toluidine blue 



Langeron 1942, 613 

 REAGENTS REQUIRED: A. ADS 12.2 Lugol (1905); B. 5% sodium thiosulfate; C. water 



100, orange G 1, erythrosin 0.2; D. 1% toluidine blue; E. 0.2% acetic acid 

 method: [sections] —> water —> a, J^ hr. -^ water, rinse -^ jS, till bleached ^ water, 



thorough wash —* C, 15 mins. — * water, rinse -^ D, on slide, 1-2 mins. — * water, 



rinse — > E, till differentiated -^ balsam, via usual reagents 

 note: Langeron {loc. cit.) says "Ce procMe, dit de Dominici, jouit en France d'une assez 



grande vogue — etc." See, however, DS 13.21 Dominici 1902, above. 



13.22 IN COMBINATION WITH OTHER DYES 



13.22 Bauer and Leriche 1934 methylene blue-eosin Y-cresyl blue 



16550, 42:1385 

 reagents required: A. water 100, brilliant cresyl blue 0.25; B. DS 13.11 Jenner 1899 



(working sol.) 

 method: Mix 4 parts blood with 1 A, as drop on slide, 2 mins. -^ smear — > dry -^ B, 



2 mins. — > water, rinse — > dry —* neutral mountant. 



13.22 Blank 1942 mercurochrome-azur-eosin 11284,27:1342 



STOCK solutions: I. water 82, 40% formaldehyde 9, abs. ale. 5, DS 13.13 Geimsa 1902 



2.25, methylene blue 0.25, sodium borate 0.5 

 reagents: required: A. 0.1% mercurochrome 220 in 25% methanol; B. stock I 5, 



water 95 

 method: [sections (nuclei may be hematoxylin stained)] — > water -^ A, \ min. — * wash 



— > B, 2 mins. ^•90% ale, till no more color comes away 



13.22 Geschickter 1930 azur-erie garnet 20540b, 5:81 



formula: water 100, azur A 0.8, erie garnet B 0.1 

 preparation: To the azur dissolved in 80 water add, very rapidly, the garnet dissolved 



in 20. Filter. 

 method: [frozen sections of fresh tissue] — > stain, 15-20 sees. — > wash — > M 10 mountant 



13.22 Houcke 1928a methylene blue-toluidine-thionin-fuchsin 



6630, 99 :786 

 preparation of stock formulas: I. Mix, without agitation, in a small graduate, 14 1 % 



acid f uchsin with 22 1 % methylene blue. Leave 24 hours. Pour liquid from viscous ppt. 



which adheres to side of graduate. Dry ppt. and dissolve in 20 methanol. II. Add 0.5 



1% acid fuchsin to 10 sat. sol. thionine. Leave 1 hour. Centrifuge. Decant and drain. 



Dry ppt. in tube. Dissolve in 10 methanol. III. Mix 11 1% toluidine blue with 5 1% 



acid fuchsin. Then treat as II. 

 working formula: water 100, stock I 2.5, stock II 2.5, stock III 2.5, 1% acetic acid 1 

 method: [sections] -^ water—* stain }i to 2 hrs. — > abs. ale, shortest possible time -^ 



balsam, via xylene 

 note: Houcke {loc. cit.) comments that the pH of the'working solution is critical, but 



the optimum, which varies both with types of tissue and fixatives employed, can only 



be established empirically. 



