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| Screening of Specimen Preparation and Fixatives of Manila Clam Ruditapes philip pinarum Haemocytes for Transmission Electron Microscopy |
| LIU Jing,YUAN Yuqing |
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Abstract Fixation is the key step of the specimen preparation for transmission electron microscopy .The aim of this paper is to investigate and compare the influence of 5 different fixatives on the ultrastructure al‐terations of haemocytes in Manila clam Ruditapes philippinarum using transmission electron microscope in order to find the proper fixative for Manila clam .The results showed that fixative affected the haemo‐cyte ultrastructure directly .The haemocytes were swelled in 2 .5% and 5% glutaraldehyde groups ,the higher in 2 .5% glutaraldehyde groups than in 5% glutaraldehyde fixative treatments ,and higher swollen in the haemocytes mitochondria than in the other organelles .No organelles swelling was found in isotonic fixative treatments ,while the inner structures of the haemocytes were not preserved very well in 5% glut‐araldehyde fixative with 7% saccharose groups and 4% paraformaldehyde‐2 .5% glutaraldehyde fixative treatments ,especially the endomembrane system .In 2% paraformaldehyde‐2 .5% glutaraldehyde fixative treatments ,the haemocytes had better ultrastructure than in other groups ,with clear outer membrane structure of haemocytes .The cytoplasmic matrix was rich and evenly distributed ,as well the ultrastruc‐ture of the organelles such as mitochondria ,endoplasmic reticulum ,Golgi apparatus and the nucleus were better preserved .From this study we can concluded that the hypotonic fixative could cause haemocytes swelling and that the mitochondria are more sensitive to the fixative osmotic pressure than other organ‐elles .Raised fixative concentration reduces the cell swelling degree ,but the high fixative concentration is not benefit to the haemocytes ultrastructure .Thus the suitable fixative is the foundation of the cell ultra‐structure preservation .
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Received: 25 May 2016
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