DS 13.12-DS 13.13 DYE STAINS OF GENERAL APPLICATION 347 



13.12 Schmorl 1928 see DS 23.33 Schmorl 1928 



13.12 Senevet 1917 5310, 10:510 



STOCK solutions: I. methylene l)lue 1, sodium borate 3, water 100; II. eosiii Y 1, water 



100 

 WOKKING solution: water 100, stock II 0.2, stock I 0.25 to 0.4 

 method: [metham>l-fixed smears] — > A, 2-3 hrs. —> wash -^ dry 



13.12 Stafford 1934 1091'.), 55:229 



REAGENTS uequired: A. Water 100, potassium dichromate 1, eosiu 1; B. DS 11.44 Good- 

 pasture (1934); C. acetone 100, abs. ale. 10 

 method: [frozen sections of formaldehyde-ale. material] —r water -^ yl, 1 sec. — > wash — > 

 B, 30 sees. -^ wash — ♦ blot on slide — > C, dropped on section till dehydrated -+ 

 balsam, via xylene 



13.12 Singh, Jaswart, and Bhattacharji 1944 see DS 11.44 Manwell 1945 (note) 



13.12 Wright 1910 Rep. Mass. Gen. Hosp., 3:1 

 FORMULA OF METHYLENE BLUE STOCK: Water 100, methylene blue 1, sodium bicarbonate 



0.5 



PREPARATION OF STOCK SOLUTION: Digest at 100°C. 1\^ hours. 



WORKING solution: methanol 80, eosin B, 0.16, stock blue 24 



method: [smear] — + stain, on slide 2 mins. -^ add water, drop by drop, till green scum 

 forms on surface, leave 2 mins. —> water, thorough wash -^ [dry] or —^ neutral 

 mountant, via acetone 



note: Wright 1910 also recommended (17510, 57:783) the dilution of the working solu- 

 tion with an equal volume of water to be used for 10 minutes. It is nowadays uni- 

 versal practice to substitute a phosphate buffer at pH 6.4 for the water used to dilute 

 the stain on the slide. 



13.13 OTHER THIAZIN EOSINATES 



13.13 Agulhon and Chavennes 1919 see DS 13.13 Pappenheim 1908 



13.13 Bohm and Oppel 1907 methylene blue-thionin-eosin 



Bohm and Oppel 1907, 114 

 REAGENTS REQUIRED: A. Water 96, 40% formaldehyde 4, methylene blue 0.3, thionin 



0.15; 5. 1% acetic acid; C. 1% eosin B 

 method: sections — > A, some mins. -^ B, tiU no more color comes away —> C, few sees. 

 —f balsam, via usual reagents 



13.13 Boye 1940 5310, 33:248 



REAGENTS REQUIRED: A. watcr 100, cosin Y 0.1; 5. DS 11.44 Stevenel 1918 

 method: [methanol-fixed smears] -^ A, 15-20 sees. — > B, flooded on slide, 45 sees. — > A, 

 till differentiated -^ rinse -^ dry 



13.13 Endicott 1945 20540b, 20:5^ 



PREPARATION OF DRY STOCK: To 250 ml. 0.8% toluidine blue add 250 0.4% eosin B. 



Filter, wash, and dry ppt. 

 WORKING solution: methanol 50, glycerol 50, dry stock 0.3 



13.13 Field 1941 see DS 23.33 Field 1941 



13.13 Geschickter, Walker, Hjort, and Moulton 1931 20540b, 6:3 



REAGENTS REQUIRED: A. watcr 60, glycerol 20, 95% ale. 20, sodium hydroxide 1.47, 

 potassium acid phosphate 0.675; B. ethylene glycol 75, 95% ale. 25, acetic acid 0.2, 

 thionine-eosinate 0.75, barium-eosinate 0.25, azur A 0.25; C. 20% glycerol in 95% 

 ale; D. diethylene glycol monobutyl ether; E. n-butyl phthalate 



PREPARATION OF B: Dissolve the barium-eosinate with heat; raise solution to boiling, 

 add thionine-eosinate. Filter solution hot. 



method: [frozen sections of fresh tissue] — > A, till required-^ B, 20-30 sees.—* C, 10 

 sees. -^ C, fresh solution, 3 sees. — > D, 10-15 sees. -^ E, 20 sees. -^ dammar 



note: Solution A should be adjusted, if necessary, to pH 7. 



