(Not included in cionyrnphfc.) Tech. E:h 



DIRECTIONS TOR MAKING FACE VII'MS AND SECTIONS OF KH4AT0DES 

 E. M. Buhrer, Section of Nematology, Beltsville, Maryland 



Satisfactory face views cannot be made from distorted, shrunken speci- 

 mens, especially spear-bearing species of Dorylamoidea, Tylenchoidea, 

 etc., in which spears are extruded if the specimens are not properly 

 killed. 



1. Kill and relax specimens by placing them in a hollow ground slide 

 filled with water and hold over a small flame. Do not overheat and 

 cook them. 



2. Fix in F.A.A. or dilute Flenmings: The latter gives a slight stain 

 which aids in locating the portions severed. 



3. Place in 1^% glycerin in 30% alcohol and evaporate to pure glycerine 

 in a small desiccator, a process which requires at least two weeks. 



U. Prepare slide with a drop of hard glycerine jelly. Do this by 



placing a small piece of jelly on the slide and carefully warming it 

 into a drop over a very small flame. Do not heat too much or bubbles 

 will form. Spread drop as thin as possible with bamboo splinter. 



5. Select the tip of a slender hair from a brush and imbed in jelly 

 drop while warm. This will act as a marker for the head and should 

 reach about half-way across the drop. 



6. Place nematode in a drop of glycerine on a thin piece of celluloid 

 and decapitate with an "eye knife." This is done under the binocu- 

 lar with the hand held firmly on the hand rest. 



7. Pick up head with a very fine bamboo splinter and hold in hand while 

 performing the next operation. 



8. Insert head into glycerine jelly at end of pointer hair and stand on 

 end, holding it in place with the bamboo splinter just touching the 

 surface of the jelly above the face. The jelly sets in a few seconds, 



9. Apply a very thin cover-glass to the drop, carefully centering it 

 before it touches. 



10. Heat the dissecting needle and carefully touch the cover-glass, melt- 

 ing the jelly \anderneath \Kitil the cover- glass is lowered to near 

 the face. Do not use too much heat in this operation or the whole 

 drop of jelly will become warm and the head fall out of place and 

 migrate. 



11. Place two small drops of candle wax just under the edge of the 

 cover-glass on opposite sides, lightly cementing it to the slide. 



