340 METHODS AND FORMULAS DS 12.33 



method: [blue nuclei (original recommends DS 11.123 Masson (1934) insufficiently 



differentiated)] -^ water—* A, 2 mins. — > water, brief rinse -^ B, 2 mins. — > water, 



brief rinse — * C, 30 sees. — * balsam, via usual reagents 

 result: cartilage, blue (from hematoxylin); other coUagens, light green; bone, brilliant 



green; epithelia and muscle, orange; erythrocytes, yellow; nervous tissue, gray. 

 note: In the opinion of the author this is the finest triple stain yet described. A detailed 



description of the use of this stain is given under 12.30 above. 



12.33 OTHER COMPLEX CONTRASTS 



12.33 Blank 1942 see DS 13.22 Blank 1942 



12.33 Flemming 1891 gentian violet-orange G 1780, 37 :249 



KEAGENTS REQUIRED: A. 1% gentian violet; B. sat. sol. (circ. 11%) orange G 

 method: [sections of Flemming 1882 or other F 1600.0000 or F 1600.0010 fixed material 

 nuclei stained by DS 11.42 method]-* water-* A, 3 hrs. — > quick wash — > B, few 

 moments — > abs. ale, till no more color comes away — > balsam, via clove oil 

 note: Johansen (Johansen 1940, 84) substitutes a saturated solution of orange G in 

 clove oil for B, above. For Stockwell's Variation see DS 13.5 Stockwell 1934. 



12.33 Hubin 1928 eosin Y-orange G-safranin 1825, 37 :25 



reagents required: A. 0.1% acetic acid; B. abs. ale. 70, water 30, eosin Y 0.1, orange 



II 0.2, safranin 0.2; C. 0.1%, hydrochloric acid in 70%, ale. 

 preparation of b: Dissolve the eosin in 50 ale. and 10 water. Add to this the orange 



dissolved in 10 water. To this add the safranin dissolved in 20 ale. and 10 water. 

 method: [blue nuclei (original specifies DS 11.122 Carazzi 1911 used on sections of 



F 5000.1040 Hollande 1911 fixed and F 5000.1010 Bouin 1897 mordanted material)] 



-* A, 4-5 sees. — * 70% ale., 5 mins. -^ B, 1-3 mins. — > C, till differentiated, 2 to 10 



sees. -^ balsam, via usual reagents 

 result: nuclei, cartilage, bone, blue; nerves and blood vessels, yellow; muscles and 



ganglia, brown. 



12.33 Johansen 1940 see DS 12.33 Flemming 1891 (note) 



12.33 Lendrum 1939 GEEP stain— and. 11431,49:590 



reagents required: A. water 50, 95% ale. 50, eosin Y 0.2, erythrosin 0.2, phloxine 0.2, 



gallic acid 0.5, sodium salicylate 0.5; B. sat. sol. tartrazine N.S. in ethylene glycol 



monoethyl ether 

 preparation of a: Add the dyes dissolved in the ale. to the other ingredients dissolved 



in water. 

 method: [sections with hematoxylin-stained nuclei] — > A, 2 hrs. ^95% ale, rinse — > B. 



on slide, till color balance satisfactory 



12.33 Lendrum 1947 11431,59:394 



REAGENTS REQUIRED: A. water 100, calcium chloride 0.5, phloxine 0.5; B. sat. sol. 



tartrazine in ethylene glycol monoethyl ether 

 method: [sections with blue nuclei] — > A, 30 mins. -* rinse -^ B, from drop bottle, till 



differentiated -* balsam, via usual reagents 



12.33 Lillie 1940 Biebrich scarlet-picro-anilin blue 1789, 29 :705 



REAGENTS REQUIRED: A. water 99, acetic acid 1, Biebrich scarlet 0.1; B. water 100, picric 



acid 1, anilin blue 0.1; C. 1% acetic acid 

 method: [sections, nuclei stained in hematoxylin] -^ water — » A, 4 mins. — > rinse — » B, 



4 mins. -^ C, 3 mins. — » salicylic balsam, via usual reagents 



12.33 Lillie 1948 see DS 13.7 LUlie 1948 



12.33 Margolena 1933 phloxine-orange G 20540b, 8:157 



reagents required: A. 0.5% phloxine in 20% ale; B. 0.5% orange G in 95% ale. 

 method: [blue nuclei] —» water —> A, 1-5 mins. —» 70% ale, thorough rinse ^ B, 

 dropped on slide, H to 1 min. — > abs. ale, till no more color comes away -^ balsam, 

 via usual reagents 



12.33 Masson 1929 metanil yellow-picro-fuchsin 11571b, 12:75 



REAGENTS REQUIRED: A. water 100, acetic acid 0.5, metanil yellow 0.5; B. 0.2% acetic 

 acid; C. 3% potassium dichromate; D. DS 11.221 van Gieson 1896; E. 1% acetic acid 



