| Summary: | 碩士 === 國立中興大學 === 獸醫病理生物學研究所 === 101 === Transmission electron microscopy (TEM) is widely utilized to tumor diagnosis through estimating the content of intracellular organelles in ultrastructural observation. However, the procedure of TEM is quite complicated comparing with general morphological technique. The target of this study was refinement of conventional TEM of endoplasmic reticulum (ER) identification and to constitute the basic data in localization of liver, pancreas and spleen.
In this study, the methods enzyme histochemistry and silver enhanced nanogold immunolabeling to localize the NADPH-Cytochrome P450 reductase and Glucose-6-Phosphatase on ER membrane of rat, dog and chicken were used to replace the conventional immunoelectron microscopy. The immunolabeling was confirmed by the anti-calreticulin, which localized on ER, nuclear envelope and nucleus. The results of enzyme histochemistry stain manifested strong signals in the liver of rat and chicken, while the results of immunolabeling stain manifested the strong signals in the liver of rat and dog. In addition, the cell in the marginal zone of spleen displayed high intense stain in both enzyme and immune labeling. In order to recognize the positive stained cells in spleen, the immunoelectron microscopy was performed and the results revealed that the signals were ER of macrophage.
In conclusion, the immune method displayed a better sensitivity and less cross-reaction than the enzyme method. Moreover, the protein expression in various tissues determined the outcome of the stain. Silver enhanced immunogold method was available to improve the tumor or disease diagnosis in veterinary medicine through the appropriate selection of target protein to localize the intracellular organelle.
|
|---|
