282 Cytoplasmic Elements (chap. 18) 



results: 



mast cells — deep blue or purple blue 

 cytoplasm — light gold-orange 

 nuceli — faint blue 



Quick Toluidine Blue Method ^^ 



fixation: any general fixative. 



solution: 



toluidine blue O, C.I. 52040 0.2 gm. 



60% ethyl alcohol lUO.O ml. 



procedure: 



1. Deparaffinize and run slides down to 60% alcohol. Remove HgCL. 

 if present. 



2. Stain in toluidine blue: 1-2 minutes. 



3. Rinse quickly in tap water. 



4. Dehydrate in acetone, 2 changes: 2-3 minutes in each. 



5. Clear in xylene and mount. 



results: 



mast cells — deep reddish purple 

 background — faint blue 



Toluidine Blue (padawer, 1959) 



fixation: formalin-alcohol preferred, 10% formalin satisfactory. 



formalin 10.0 ml. 



95% alcohol 90.0 ml. 



solutions: 



toluidine blue O, C.I. 52040 0.05 gm. 



ether 181 i^ 50.0 ml. 



distilled water 50.0 ml. 



Dissolve dye in water, add ether 181. Filter. Use within 24 hours. 



procedure: 



1. Deparaffinize and transfer into absolute methyl alcohol, 2 changes: 

 10 minutes each. 



2. Transfer to ether 181, 50% aqueous: 10 minutes. 



3. Stain in toluidine blue: 20-60 minutes. 



4. Dehydrate in ether 181, 50% aqueous: 10-15 minutes. 



5. Dehydrate in ether 181, 100%, 2 changes: 20 minutes each. 



6. Clear in xylene and moimt. 



" Personal communication from Miss Marlies Natzler, U.C.L.A. 



^* Tetra methylene glycol ether, Ansul Chemical Co., Marinette, Wis. 



