DS 12.222-DS 12,3 DYE STAINS OF GENERAL APPLICATION 



329 



12.222 Male 1924 fiichsin-martius yellow 11035,42:455 



formula: water 80, 95% ale. 20, acid fuchsin 0.6, inaitius yellow 0.8 



12.222 Masson 1911 saffron-cnjlhrosin 6630,70:573 



REAGENTS REQUIRED: A. 1% erythrositi ; B. wnfcr 100, safTron 2, 5% tannin 1, 40% 



fonualdehyde 1 

 PREPARATION OF B: Extract the .saffron in (he water 1 hour OO^C. Filter. Add other 



ingredients to filtrate. 

 method: [blue nuclei]-* water -> A, 5 mins. ^ water, quick rinse -> 70% ale, few 



sec. -> water, thorough wash -* B, b mins. -^ blot -> abs. ale, flooded over slide, 



till dehydrated —> balsam, via xylene 

 note: Langeron 1942 (p. 596) substitutes eosin B for erythrosin in the above. 



12.222 Semichon 1920 methyl blue-eosin-victoria yellow 



5401,45:73 

 REAGENTS REQUIRED: A. Water 100, methyl blue 0.04, eosin Y 0.2, victoria yellow 0.1 

 method: [Ijlue nuclei]^ water -^ A, overnight^ drain -> abs. ale, till differentiated 



— > xylene — > balsam 

 result: horn, hair, chitin, yellow; cartilage, blue; other tissues, orange, 



12.222 Squire 1892 fuch sin-orange Squire 1892, 42 



formula: water 80, 95% ale. 20, acid fuchsin 0.3, orange G 2.0 

 method: as Gray 1952 above 



12.222 Sziitz 1912 polychrome alizarin 23632,29:289 



REAGENTS REQUIRED: A. 5% aluminum acetate; B. sat. ale. sol. alizarin red S 1, water 100 

 method: [hematoxylin-stained sections of F 2300.1000 Sziitz 1912 fixed material]—* 



water — > ^, 5 hrs. — » rinse -^ B, 5 hrs. -^ wash — > balsam, via usual reagents 

 result: nuclei, blue; cytoplasm, varying shades of red, cytoplasmic inclusions being 



generally very darkly stained. 



12.3 Complex Contrast Formulas 



These complex contrast formulas are 

 specifically designed to differentiate the 

 cytoplasm into various histological com- 

 ponents. They are more widely used in 

 embryology than in general histology, and 

 the first two classes, here given, are all 

 developed from the original discovery of 

 Mallory 1904 (11189, 5:15) that a solution 

 of phosphomolybdic or phosphotungstic 

 acid will remove acid fuchsin from col- 

 lagen while allowing it to remain either in 

 the muscle or in the nuclei. Mallory 's orig- 

 inal method, however, involves staining 

 the nuclei with acid fuchsin as an inherent 

 part of the technique, and is therefore 

 given under the subheading DS 13, in 

 which combined nuclei and plasma stain- 

 ing techniques are dealt with. The group 

 here discussed is commonly associated 

 with the name of Masson, and the term 

 "Masson's Trichromic" is widely applied. 

 These methods require prior staining of 

 the nuclei with hematoxylin and the sub- 

 sequent staining of the connecting and 

 supporting tissues in accordance with Mal- 



lory's principle. This stains nuclei a color 

 from blue to deep purple, and they are 

 thus distinguished far more clearly from 

 the background cytoplasm than are those 

 stained by the original method. All of 

 these phosphotungstic and phosphomo- 

 lybdic methods are excellent. 



The second class of complex contrast 

 formulae are those employing phosphomo- 

 lybdic or phosphotungstic acid reaction 

 with other dyes than fuchsin, while at the 

 same time retaining the differential nu- 

 clear stain. Of these, Patay 1934 is a very 

 nearly fool-proof and brilliant method of 

 triple staining. Indeed the whole class of 

 stains, with tlie exception of Fleming 1891, 

 are of recent origin, and at the present 

 time are the most popular group of coun- 

 terstains which have yet been developed. 

 Though they are customarily employed in 

 zoological technique, no one who has tried 

 them in botanical technique has ever re- 

 gretted it. 



The final class of complex contrast for- 

 mulas contains only those mixtures which 

 cannot otherwise be classified. Among 



