Tech. E:3 



^$!. ethyl alcohol 15 or 20 ml. 

 Glacial acetic acid 1 ml. 



Formalin (UO/^ f onnaldehyde) 6 ml. 



Distilled water UO ml. 



After relaxing by gentle heat, the nematodes are covered with the 

 fixative and left there for 2[i-U8 hours. A slight staining of the 

 nematodes can be obtained if a few drops of saturated picric acid are 

 added to the fixative. 



Lacto-phenol Solution 



Melted phenol 3 parts 



Lactic acid 1 part 



Glycerine 2 parts 



Water 1 part 



Cj'st-forms and female root-knot nematodes can be mounted directly from 

 water or formalin into lactophenol solution for examination of cuticu- 

 lar details. The cover-glass need not be sealed, as this material 

 evaporates at a very slow rate. 



Permanent Slides 



Slides of soil and plant nematodes capable of lasting many years can 

 be made but are usually, and perhaps wrongly, considered to involve 

 too much trouble for routine work. However, specimens of taxonomic 

 value should be made into permanent mounts. Two methods for making 

 such slides are presented. The first method requires a lengthy lapse 

 of time to allow dehydration of the nematodes to occur and is the 

 method developed by G. Thome of the Section of Nematology, Nematodes 

 on slides prepared in this manner over 30 years ago are still in an 

 excellent state of preservation. The second method is popular because 

 of being quicker, due to a different manner of dehydration. 



Special slides include mounts made of nematodes for examining the en 

 face aspect of nematodes, cross-sectional views, and tail mounts, The 

 method of Buhrer given in this manual can be used for making permanent 

 mounts of cross-sections, nematode tail sections, as well as for head 

 mounts. The methylcellulose method is used only for rapid preparation 

 of mounts which are not permanent. 



Miscellaneous Staining Methods 



Intra-vital stains of various kinds are of value in water mount slides 

 of most of the nematodes, except those of the Tylenchoidea and Aphelen- 

 choidea groups. Anilin blue .WS (Syn: cotton blue, water blue, china 

 blue, Poirrier's blue) is a very useful stain which aids in the resolu- 

 tion of cuticular structures and markings and for locating amphidial 

 and phasmidial openin.';s. A small drop of saturated stain in vjater is 

 added directly to living or fixed specimens on the slide. 



