348 Histochemistry (chap. 21) 



3. 95% acetone: 1 minute. 



4. 85% acetone: 1 minute. 



5. 50% acetone, 1 minute. 



6. Water: 1 minute. 



7. Incubate, room temperature: 3-20 minutes. 



8. Wash in tap water: 2 minutes. 



9. Hematoxylin: 3 minutes. 



10. Wash, tap water: 5 minutes. 



11. Glycerol jelly. 



results: 

 sites — red 



comments: 



Nachlas and Seligman (1949) prefer b over a-naphthylacetate, say- 

 ing that it gives greater brilliance of reaction. 

 Wachstein and Wolf {1958) incubate 30-90 minutes in: 

 1% propylene glycol (in 0.2M phosphate buffer 



at pH 6.9) 40.0 ml. 



1% naphthol-AS acetate in acetone 0.4 ml. 



fast blue 55 salt (C.I. 37175) 80.0 mg. 



Lipase (gomori, I950E, 1952) 



fixation: chilled acetone: 24 hours (2mm. sections: prechill tissue to 

 lessen shrinkage; trim thinner when tissues have gained some con- 

 sistency). Embed by double embedding, page 339. Temperature 

 should not exceed 58 °C. If sections refuse to flatten completely, blot 

 onto slides, dry at room temperature, then melt in paraffin oven. 



solutions: 

 Substrate: 



Tween 60 (or product :^81, Onyx Oil and 



Chemical Co., or G2151, Atlas Powder Co.) . . 5.0 gm. 

 distilled water 100.0 ml. 



Tris buffer, pH 7.2-7.4: 



maleic acid 29.0 gm. 



tris (hydroxymethyl) aminomethane 30.3 gm. 



distilled water 500.0 ml. 



add charcoal 2.0 gm. 



Shake, let stand 10 minutes, and filter. 



