DS 12.33-DS 13 DYE STAINS OF GENERAL APPLICATION 341 



method: [sections with blue nuclei (original requires DS 11.111 R^gaud 1910)] -^ water 

 -^ A, 5 mins. — > B, rinse — > C, 5 mins. — >• D, poured on slide still wet with C, 2 mins. 

 —y E, till yellow clouds cease -^ salicylic balsam, via usual reagents 



result: collagens, clear red. 



12.33 Masson 1911 erythrosin-saffron 6630, 70:573 



REAGENTS REQUIRED: A. water 100, erythrosin 1, 40% formaldehyde 0.25; B. water 100, 

 saffron 2, 40% formaldehyde 1, 5% tannic acid 1 



PREPARATION OF B : Boil saffron 1 hour in water. Cool. Filter. Add other ingredients. 



method: D-Aue nuclei] -^> ^, 5 mins. -^ water, quick rinse— » 70% ale, till collagens 

 colorless -^ water, quick rinse -^ B, 5 mins. -+ water, rapid rinse — > abs. ale, mini- 

 mum time possible — » balsam, via xylene 



result: nuclei, blue; collagens, yellow; muscle, red. 



12.33 Maximow 1909 eosin Y-azur II 23632,26:177 



STOCK FORM I las: I. 0.1% cosiu Y; II. 0.1% azur II 

 WORKING solution: water 100, stock I 10, stock II 10 

 method: [sections with chromatin stained in very dilute DS 11.122 formula] -^ water, 



24 hrs. -> stain, 12-24 hrs. -^ 95% ale, quick rinse -* abs. ale, till differentiated -^ 



neutral balsam, via X3'lene 



12.33 Millet 1926 acid fuchsin-martius yellow 4285a, 3:2 



REAGENTS REQUIRED: A. 5% acid fuchsin in 40% ale; B. 5% martins yellow in 40% ale. 

 method: [blue nuclei] -* A, 5 mins. at 30°C. — » 40% ale, till no more color comes away 



— > fi, 5 mins. -^ balsam, via usual reagents 

 note: This method was developed for insect histology, for which it is excellent. 



12.33 Pianese 1896 malachite green-acid fuchsin-martius yellow 



2526, 1:193 

 formula: water 75, 95%, ale 25, malachite green 0.25, acid fuchsin 0.05, martius yellow 

 0.005 



12.33 Reinke 1894 gentian violet-orange G 1780, 44:262 



formula: a. sat. sol. {circ. 1%) gentian violet 25, sat. sol. {circ. 11%) orange G 0.2, 



water 75 

 note: Use after safranin nuclear staining (DS 11.421) and differentiate with clove oil 



12.33 Scriban 1924 picro-fuchsin-brilliant green 6630,90:531 



reagents required: A. water 100, picric acid 0.5, acid fuchsin 0.1; B. 60% ale 100, 



picric acid 0.2, brilliant green 0.1. 

 method: [sections with DS 11.111 stained nuclei] -^ thorough wash-* A, 3-4 sees. -» 



abs. ale, wash -^ B, 3-4 sees. -^ abs. ale, wash -^ balsam via usual reagents 



12.33 Stockwell 1934 see DS 13.5 Stockwell 1934 



12.33 de Winiwarter 1923 see DS 12.33 de Winiwarter and Sainmont 1908 



12.33 de Winiwarter and Sainmont 1908 crystal violet-orange G 



23632, 25:157 

 reagents required: A. 1% crystal violet; B. sat. sol. (circ. 11%) orange G; C. 0.1%, 



HCl in abs. ale 

 method: [sections of F 1600.0000 or F 1600.0010 fixed material; nuclei stained by 11.42 

 method] -^ water -» ^, 24 hrs. -* brief rinse -y B, 1 min. -* C, 2 to 3 hrs. -* clove 

 oil, till differentiated -^ balsam 

 note: de Winiwarter 1923 (1825, 32 :329) recommends a process which differs only in the 

 substitution of 0.2% orange G for B, above. 



13 COMPLEX TECHNIQUES INVOLVING BOTH NUCLEAR 

 AND PLASMA STAINING 



This is a large class of staining tech- elements of the background are differ- 



niques, involving those methods in which, entially stained. They are divided broadly, 



by a series of successive and interlocking for purposes of this work, into seven 



operations, both the nuclei and all the classes, of which the first two (DS 13.1 



