SECTION TWO 



unstained sections, two troublesome features are frequently met 

 with; i.e. (i) a halo of light around isolated cells and cellular 

 structures and, (ii) insufficient contrast, which is particularly 

 troublesome in studying epithelial sheets, cell boundaries being 

 difficult to discern. Light filters of different wavelengths were 

 tried without success, but the staining technique described above, 

 which according to the author was discovered by an accident, was 

 found to increase contrast and eliminate halo-ing. 



(b) Structures well defined by this technique, which is stated 

 to be of no value for conventional microscopy, include terminal 

 bars, epithelial tonofibrils, brush borders, striated borders, 

 zymogen granules, cytoplasmic vacuoles, filamentous mitochondria 

 of renal tubules, and according to the author, many others too 

 numerous to mention. 



(c) For further information readers should consult the original 

 paper, and J. A. Green's Luxol fast scarlet method (1957) which 

 is described on page 280 of this book. 



(d) I would suggest that Luxol fast orange, Luxol fast red, 

 and Alcian blue might also be tried out along the same lines as 

 Luxol fast scarlet C and Luxol fast blue. 



Reference: Green, J. A. (1956). 



LUXOL BRILLL\NT GREEN BL 

 A nuclear stain for phase-contrast microscopy 



Solutions required: 



A. David's mordant 

 Potash alum 

 Mercuric chloride 

 Tannic acid 

 Distilled water 



B. Luxol brilliant green BL 

 Alcohol, 95% 



5gm 



I -25 gm 



4gm 



90 ml 



0-25 gm 

 100 ml 



Technique : 



I. Fix material in 10% formalin and embed in paraffin wax as 

 usual. 



275 



