DS 13.31 



DYE STAINS OF GENERAL APPLICATION 



355 



Sections are passed directly from dis- essential that the slide be passed from 



tilled water into the stain, in which they 

 may remain from overnight to about 24 

 hours. Overstaining does not normally 

 occur. 



Each slide must be removed separately 

 from the stain and rinsed briefly and 

 rapidly in 95% alcohol. It will be observed 

 that in the alcohol color clouds rise from 

 the sections, and that there is then a brief 

 period when these color clouds cease. If 

 the slide is then left for a longer period in 

 alcohol, further stain will commence to 

 leave it. To secure satisfactory slides, it is 



alcohol to the carbon-xylene, which is 

 recommended for clearing, at the exact 

 moment when the primary color clouds 

 cease to leave. The purpose of the carbol- 

 xylene is to permit complete dehydration, 

 even though the length of time in alcohol 

 has not been sufficient to remove the 

 whole of the water from the section. The 

 carbol-xylene must be thoroughly re- 

 moved with pure xylene before the slide 

 is mounted in balsam, or the latter will 

 inevitably turn brown. 



13.31 METHYL GREEN IN COMBINATION AVITH PYRONIN 



13.31 Flinn 1939 see DS 23.219 Flinn 1939 



13.31 Grosso 1912 16059, 4:41 



formula: water 100, sat. sol. (arc. 9%) pyronin 5, sat. sol. {circ. 5%) methyl green 3.5, 

 sat. sol. {circ. 11%) orange G 3.5 



method: water —> stain, 30 sees. — > abs. ale, till differentiated^ n-projiyl ale, if de- 

 hydration insufficient -^ neutral mountant 



result: nuclei, green; plasma, red, orange, and yellow. 



13.31 Langeron 1942 Langeron 1942, 615 



STOCK solutions: I. water 100, methyl green 4, phenol 5; II. water 100, pyronin 4, 



phenol 5 

 REAGENTS REQUIRED: A. stock I 50, stock II 50; B. ale. 50, acetone 50; C. aniyl alcohol 

 method: distilled water — > A, 15 mins. at 50°C. -^ distilled water, rinse -^ B, till differ- 

 entiated -^ C, till dehydrated -^ balsam, via toluene 

 result: good differential staining of blood cells and glandular cell inclusions. 



13.31 Lipp 1940 see DS 23.214 Lipp 1940c 



13.31 Pappenheim 1901 1780a, 155:427 



STOCK solutions: I. water 100, phenol 0.25, methyl green 1; II. water 100, phenol 0.25, 



pyronin 1 

 WORKING formula: stock A 30, stock B 70 

 method: water ^^ stain, 5-10 mins. -^ water, quick rinse —> neutral mountant, via 



acetone 

 result: nuclei, violet; lymphocytes and plasma cells, red; other tissues, orange and 



green. 

 note: For bacteria see DS 23.221 Saathof 1905. 



13.31 Sandiford 1937 11431,45:467 



formula: water 75, glycerol 20, 95% ale. 5, phenol 1.5, methyl green 0.15. i)yn)iiiii 0.5 



13.31 Scott and French 1924 13685, 55:337 



formula: water 80, glycerol 16, ahs. ale. 4, phenol 1.6, nietliyl green 0.8, pyronin 0.2 



13.31 Unna 1910 test. 1928 Gatenby and Cowdry cit. Gandletz 



Gatenby and Cowdry 1928, 176 

 formula: water 100, phenol 0.5, glycerol 20, abs. ale. 2.5, pyronin 0.25, methyl green 



0.15 

 preparation: Griiul the dyes with the ale in a mortar. Heat glycerol to 50°C. and add 



to mortar in small jxtrtioris while frriiuling. Dissolve phenol in water and use this to 



wash out mortar with small successive doses. 

 method: water -* stain, 10 mins. 30°C. -> water, rinse — » abs. ale till differentiated — ' 



balsam, via usual reati;eiits 

 note: For adaptation for bacterial staining, see DS 23.221 Saathof 1905; for staining 



fungi in skin sections see DS 23.32 Unna 1929. 



