Tech. E:9 



include Chlorazol Black, Fast Green, Bismark Brown, Purpurin K, and 

 B, Scarlet. Orange G is a good general stain. A stain that the 

 writer has found useful is Van Giesen's RLcro-Fuchsin. Some very 

 interesting differential staining may often be obtained with this 

 medium, with bright colors and very little danger of overstaining. 



Controlling temperatures while using this mounting method may appear 

 to be a problem if constant temperature equipment is not available. 

 If possible, reduce the peak temperature of your hot plate below the 

 boiling point of water, (preferably around 60 - 70° C. ). Five drops 

 of one of the mixtures are about sufficient to fill the shallow de- 

 pression of a hanging-drop slide. Transfer the specimens to this slide. 

 Place the slide on the hot plate with the tip of your index finger on 

 top of the slide at one end. When the temperature is too hot for your 

 finger, take the slide off the plate. 



There is no present indication that this method gives better results 

 than original Rapid Method (Baker 19^3 )> hut its main value may lie in 

 the fact that fewer transfersare necessary. 



PERMNETJT NEllATODE ^DUNTS IN GLICERII\IE 

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



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

 nent mounts as described by Dr. Seinhorst diiring his visit in 1957 to 

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

 tology Project (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. Seinhorst' 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-^2° C). The 

 progress of this step can be observed under the dissecting scope 

 by using a cavity slide containing a small quantity of vjater 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 5^ formaldehyde can be used as fixatives. Cover 

 dish to retard evaporation. 



1. If fixative used is at ^0-52° C, keep nematode in it for 12-21; 

 hours. 



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

 2li-li8 hours or longer. 



