Tech. D:ii 



FIXATION TOR N^xIjVTIZR]} PXJ')TS 



(Araberger technique; Flentrning Medium fixative; acetic acid-glycerine 

 clearing ; pure glycerine mounting . ) 



Section of Nematology, Eeltsville, Maryland 



Flemming Medium fixative: 1% chromic acid solution 375 cc. 



glacial acetic acid 25 cc. 



distilled water 50 cc. 



1 gram osmic acid (in sealed vial) 

 Mix the chromic acid solution, the glacial acetic acid, and the distilled 

 water in a graduate. Pour about 25 to 50 cc. of the solution into a 

 double- stoppered bottle that has been thoroughly cleaned. Avoid getting 

 any oil — even finger marks — where the fixative will touch. Remove the 

 labels from the outside of the osmic acid vial and clean its surface 

 with alcohol, again avoiding any finger marks. Drop vial into the 

 double- stoppered bottle, and break it by stamping down with a cleaned 

 heavy glass rod, thereby releasing the osmic acid. Immediately pour on 

 the rest of the chromic acid-acetic acid solution. 



Store bottle in a dark container, away from light. 



Place washed roots in the fixative, using only as much fixative as will 

 cover the roots, in a small, well-corked bottle. Allow to stand 1, 2, 

 or 3 days, depending on size of roots and color desired. 



Wash in running water 1, 2, or 3 days. 



Transfer by steps (20^, 30^, UO^i alcohol, several hours to 1 day each) 

 to S0% alcohol. For this a bottle or closely- covered glass dish pay be 

 used. 



Transfer roots to an open Syracuse watch-glass or Petri dish, depending 

 on the quantity of roots in the lot. 



Make up a solution of \ glacial acetic acid and \ pure glycerine; keep 

 in a drop-bottle. 



Drop acetic-glycerine solution into dish containing roots gradually 

 during the period of a week or less, depending upon temperature and 

 humidity conditions, allowing the alcohol to evaporate at the same time 

 that the acetic-glycerine is penetrating the roots. For a small quantity 

 of roots, well covered by the S0% alcohol, in a Syracuse watch-glass, 

 the process may be begun with 5 to 7 drops of acetic- glycerine in the 

 morning, the same amount in the afternoon, then gradually increasing the 

 amount dropped during the next few days. 



As each quantity of acetic-glycerine is added, agitate or stir the dish 

 to facilitate uniform mixing. 



