Fdl S/difiing 255 



solutions: 



Alcohol-Twcen solution: 



Tween 40 - 1 .0 ml. 



distilled water ..." 70.0 ml. 



absolute alcohol to make 100.0 ml. total 



Oil red O solution: 



Add to above, 250 mg. Oil red O, C.I. 26125. Incubate at 60°C for 

 not less than 24 hours well sealed to prevent evaporation. Agitate oc- 

 casionally. Cool, filter under vacuum. 



Hematoxylin, see page 125. 



procedure: 



1. Frozen sections, 15 microns; place in water. 



2. Transfer to alcohol-Tween Avithout dye: 10 minutes, agitate. 



3. Stain at least 4 hours: may be overnight. 



4. Differentiate in alcohol-Tween: 10-15 minutes. 



5. Wash in distilled water. 



6. Stain in Mayer's hematoxylin (or like): 2-3 minutes. 



7. Wash in water, and blue in Scott's solution. 



8. Wash in tap water: 5 minutes. Mount in gum syrup, glycerol jelly, 

 or Farrant's medium. Ring cover glass for permanency. 



results: 



fat — brilliant red 

 nuclei — blue 



comments: 



This method rarely deposits precipitate on the sections. The author 

 always uses it in preference to the former method. 



Sudan IV or Sudan Black B (chiffelle and putt, 1951) 

 fixation: 10% formalin, no alcohol. 



SOLUTION : 



Dissolve 0.7 gm. Sudan IV, C.I. 26105, or Sudan black B, C.I. 26150, 

 in 100 ml. propylene or ethylene glycol. Add small amounts at a time, 

 and heat to 100-1 10°C, stirring, for a few minutes. Do not exceed 

 1 10°C or a gelatinous suspension is formed. Filter hot through What- 

 man #2 paper, and cool. Filter again through fritted glass filter with 

 aid of suction, or glass wool with vacuum. 



^ .\tlas Powder Co.. AVilmington, Delaware. 



