348 METHODS AND FORMULAS DS 13.13 



13.13 Giemsa 1902 23684,31:429 



formula: 0.05% eosin 95, 0.8% azur 5 



note: This is the original formula for Giemsa's stain, given in a footnote to the paper 

 cited. From this time on almost any eosin-aznr mixture has been referred to as 

 Giemsa. The formulas of Slider and Downey 1929 and Gatenby and Cowdry 1928 are 

 referred to by their publishers as Giemsa's stain. When reference is made in the present 

 work to DS 13.13 Giemsa 1902 the solution of SHder and Downey 1929 should be em- 

 ployed urdess dry powder is specified. Kopel 1945 (4349, 25 :61) substitutes ethanol for 

 methanol in preparing the working solution. 



13.13 Giemsa see also DS 13.13 Langeron 1942, Slider and Downey 1928, Gatenby and 

 Cowdry 1928 



13.13 Grosso 1914 see DS 21.3 Grosso 1914 



13.13 Haynes 1926a 20540b, 1:68 



REAGENTS REQUIRED: A. 1.5% azur II or azur C; B. sat. sol. ethyl eosin in clove oil 

 method: [water] —> a, 5 mins. — > abs. ale. quick rinse ^ B, 30 sees. —> balsam, via 

 xylene 



13.13 Haynes 1926b 20540b, 1:107 



REAGENTS REQUIRED: A. 2.5% phloxine; B. 0.1% azur I; C. ADS 22.1 Wolbach 1911 

 method: [sections] -^ water — » A, 15 mins. -^ water, thorough wash -^ B, 30 mins. -^ 



water, thorough wash — * C, from drop bottle, till differentiated — > abs. ale, minimum 



possible time — > balsam, via xylene 



13.13 Kingsley 1935 20540b, 10:127 



STOCK solutions: I. water 50, methanol 10, glycerol 10, buffer pH 6.9 30, methylene 



blue 0.130, azur A 0.020; II. acetone 70, methanol 20, glycerol 10, methylene violet 



0.026, eosin Y 0.090 

 reagents required: A. stock I 50, stock II 50; B. 0.015% acetic acid; C. acetone 60, 



acetic acid 0.005, eosin Y 0.0005; D. n-butyl ale. 60, eosin Y 0.001 

 METHOD FOR SMEARS: [air-dried smear] -^ methanol 3^^ to 1 min. -^ dry -^ A, 5-8 mins. 



— > water, thorough wash — * dry 

 METHOD FOR PARAFFIN SECTIONS: Water -^ A, flooded on slide, 9-10 mins. -^ water, 



thorough wash -^ C, rinse -^ D, rinse — > balsam, via xylene 

 TEMPORARY METHOD FOR FROZEN SECTIONS: [blot section to slide] — > A, flooded on slide, 



2-3 mins. — » wash — » [examine] 

 - PERMANENT METHOD FOR FROZEN SECTIONS: [blot sectiou to slide] — > A, 4-5 mins. (fresh) 



or 8-10 mins. (fixed) -^ [thence as for paraffin sections] 

 note: The application of this stain to a blood smear is described under DS 13.10 above. 



See also DS 21.42 Kingsley 1937. Ritchie 1941 {Tech. Bull, 2 :157) prefers, for smears, 



to dilute the A stain 8:5 with water. 



13.13 Kingsley 1937 see DS 21.42 Kingsley 1937 



13.13 Langeron 1942a Giemsa for wet smears — aucl. Langeron 1942, 583 



REAGENTS required: A. Water 100, potassium iodide 2, ADS 12.2 Lugol (1905) 3; B. 



0.5% sodium thiosulfate; C. water 97, DS 13.13 Giemsa 1902 3; D.\% monosodium 



phosphate 

 method: [smears fixed in F 3000.0000 Schaudinn 1893 or F 3700.0010 Zenker 1894, 



24 hrs.] — > rinse -^ A, 5 to 10 mins. — > rinse — » B, 10 mins. -^ running water 5 mins. 



-* distilled water — > C, 30 mins. -^ C, fresh solution, 12 hrs. -+ D, if differentiation 



required — > neutral mountant, via graded acetone-xylene mixtures 



13.13 Langeron'l942b Giemsa for sections — auct. Langeron 1942, 583 



REAGENTS required: A. 70% ale. 97, ADS 12.2 Lugol (1905) 3; B. 0.5% sodium thio- 

 sulfate; C. water 97, DS 13.13 Giemsa 1902 3 

 method: [5 M paraffin section of F 3000.0000 Schaudinn 1893 or F 3700.0010 Zenker 1894 

 material, dewaxed and brought to water] -^ A, 20-30 mins. -^ rinse -^ B, 10 mins. 

 -^ tap water 5 mins. -^ distilled water — > C, 30 mins. — > C, fresh solution, 2 to 12 

 hrs. — ♦ balsam, via graded acetone-xylene mixtures 

 note: Langeron (loc. cit. p. 751) also refers to this as Wolhach's method; but see DS 13.13 

 Pappenheim 1908 (note) and DS 23.12 Wolbach 1919. 



