DS 13.22-DS 13.3 DYE STAINS OF GENERAL APPLICATION 353 



13.22 Houcke 1928b meOnjlene bhie-rhodamine B 6630, 99 :788 



formula: sat. sol. (circ. 5%) methylene blue in 95% ale. 3, sat. sol. aniline 27, 0.5% 



rhodamine B 70 

 method: [section] —> water —> stain, 2-3 hrs. — » abs. ale, minimum possible time-* 



dammar via xylene 

 result: chromatin, blue violet; nucleoli, red; collagens, orange; muscle, deep orange; 



erythrocytes, bright red. 

 note: For tissues recently fixed Houcke recommends that the aniline solution be diluted 



1 : 1 with water. 



13.22 Langeron 1942a poli/chrome methylene blue-orcein 



Langeron 1942, 612 

 reagents required: A. water 100, polychrome methylene blue 1; B. 70% alcohol 100, 



orcein 0.25 

 method: Distilled water — > A, 5 mins. -^ wash — > B, 5 mins. — > abs. ale, least possible 



time — * balsam, via xylene 

 result: nuclei, blue; connective tissue, red; other structures, polychrome. 



13.22 Langeron 1942b polychrome methylene hlue-lannin-oramje 



Langeron 1942, 612 

 REAGENTS REQUIRED: A. Water 100, polychrome methylene blue 1; B. DS 11.44. Unna 



1892 

 method: Distilled water -^ A, 5-15 mins. -^ wash -^ B, till no further blue removed — » 



rinse — > balsam via usual reagents 

 result: nuclei in mitosis, bacteria, some connective tissues, blue; resting nuclei, most 

 other structures, orange. 



13.22 Kull 1913 ioluidine blue-aurantia-acid fuchsin 766,45:153 



reagents required: A. sat. sol. aniline 100, acid fuchsin 20; B. 0.5% toluidine blue; 



C. 0.5% aurantia in 70% ale. 

 method: [sections] -^ water -^ A, 1 min. warmed to steaming —> cool—* water, quick 



rinse — * B, 1-2 mins. — * water, quick rinse -^ C, till sufficient red extracted, 20-30 



sees. -^95% ale, till differentiation complete —* balsam, via usual reagents 

 note: Though originally intended for the demonstration of mitochondria, this is an 



excellent general-purpose stain. For the mitochondria technique see DS 22.2 Kull 1914. 



13.22 Masson test. Langeron 1942 thionin-picric acid Langeron 1942, 613 



reagents required: A. DS 11.44 NieoUe (1942); B. 0.2% acetic acid; C. sat. sol. picric 



acid in toluene 

 method: water -^ a, 15 mins. — » wash — > 5, till differentiated -^ abs. ale. till de- 

 hydrated —* toluene — > C, till green —* balsam, via toluene 

 result: nuclei, bacteria blue; other tissues, yellow, blue, or green. 



13.22 Rhamy 1930 rnethylene blue-eosin Y-magenta 11284, 15:490 



stock, formulas: I. sat. sol. {circ. 6%) magenta in abs. ale; IL sat. sol. {circ. 45%) 



eosin Y in water; III. sat. sol. {circ. 1.5%) methylene blue in abs. ale 

 working formula: 30% ale 100, stock I 4, stock II 5, stock III 15 

 method: [sections]—* 70% ale—* stain, 5 mins.—* abs. ale, till differentiated—* bal- 

 sam, via usual reagents 



13.22 Unna lest. 1928 Schmorl methylene blue-orcein Schmorl 1928, 76 

 reagents required: A. DS 11.44 Unna 1892; B. 1% orcein in 95% ale 

 method: [celloidin sections of ale fixed material] -* water—* A, 10 mins. — * thorough 

 wash — * blot —> B, 15 mins. -^ balsam, via bergamot oil 



13.3 Techniques Employing methyl green combinations, it has the dis- 



Methyl Green as the advantage of being very sensitive to alka- 



NucLEAR Stain lies — so much so that extreme care must 



Methyl green in combination with py- be taken either to provide a perfectly 



ronin has been long recognized as an excel- neutral mounting or to use some acid 



lent stain for smears, but, like all other mountant such as salicylic balsam. The 



