CRYOFIXATION AND CRYOSUBSTITUTION AS A METHOD FOR PREPARING MELOIDOGYNE-JAVA']JAVANICA EGGS FOR ELECTRON-MICROSCOPY

Transmission electron microscopy (TEM) of nematodes eggs is difficult because the lipid layer of the egg shell prevents penetration of the chemicals used for fixation and embedding. Conventional chemical fixation of intact eggs does not allow observation of the internal egg components as they are damaged during processing. Cryofixation and cryosubstitution was used as an alternative method for the preparation of intact nematode eggs for sectioning and resulted in sections of eggs that were well fixed and free from the artefacts caused by chemical fixation. The integrity of embryos and larvae within eggs was maintained. The nematode eggs can now be studied as an intact system by using this method.