Tech« E: 10 



PERMANENT NEMATODE MOUNTS IN GLYCERIN 

 Based on a procedure of Dr. J. W. Selnhorst 



The following instructions are based on a procediare for making perma- 

 nent mounts as described by Dr.. Selnhorst during his visit in 1957 to 

 the southeastern states as a function of the Southern Regional Nema- 

 tology Pro;)ect (S-19) . Complete details of this method are to be 

 published in the near future. The instructions given here are to be 

 considered only as preliminary and not necessarily according to 

 Dr. Selnhorst 's technique in all details. The method, or modifica- 

 tions of it, are in use in several laboratories in the South and 

 proving quite satisfactory for mounting most nematode species, 



A. Relax and kill the nematodes with gentle heat (50-52*^0.) The 

 progress of this step can be observed xmder the dissecting scope 

 by using a cavity slide containing a small quantity of water and 

 the specimens. 



B. Transfer the nematodes to a Bureau of Plant Industry watch glass 

 or other suitable dish containing fixative. FA, FAA, Kahle's 

 solution, TAF, or S% formaldehyde can be used as fixatives. Cover 

 dish to retard evaporation. 



1. If fixative used is at 50-52°C., keep nematode in it for 12- 

 2\x hours. 



2. Or, if fixative used is at room temperature, keep nematodes 

 for 2U-U8 hours or longer, 



C. Transfer fixed nematodes into methyl alcohol solution (glycerin 

 S% in methyl alcohol) in a Bureau of Plant Industry watch glass 

 or other container. Place on hot plate regulated to 60-6U°C, 



D. Keep containers on a hot plate at 60-6U°C. until the methyl alcohol 

 has evaporated (about 30 minutes). Collapsing, if it occurs in 

 some specimens, can be corrected by leaving container on the hot 

 plate for a longer time. 



E. Transfer containers with the specimens to a dessicator for at ]e ast 

 one day. 



F. Mount specimens in anhydrous glycerin. 



