Effect of Ultrasonic Bath in Preparing Specimens for Transmission Electron Microscopy

전자현미경 시료 제작에 있어 Ultrasonic bath의 유용성에 관한 연구

The present study was carried out to investigate the effect of ultrasonic bath in tissue preparation for transmission electron microscopy. The method used standard reagents and media, and employed ultrasonic bath agitation to accelerate fluid exchange. The liver kidney, stomach and cardiac muscle tissues of male Sprague-Dawley rats were used for the experiment, and the experimental design was divided into 4 groups; The control group using rotators (Traditional method, 1,625 mins) and the three experimental groups using ultrasonic bath (UB) in the primary fixation through the infiltration processes (UB I; 62.5 mins, UB II; 125 mins, UB III; 250 mins). The results were as follows; 1. In the control group, tissues were easily sectioned, and showed well preserved intact membranes, and cell organelles such as mitochondria, lysosome, peroxisome, rough endoplasmic reticulum and smooth endoplasmic reticulum. 2. In the UB treated group I, tissues showed holes due to the inadequate removal of both water and fluids used in the dehydration process. Also the mitochondria of cell organelles, especially, showed swollen intracristal spaces and dense matrices due to poor fixation. 3. In the UB treated group II, tissues showed good preservation of cell organelles and specimen slice sections. Also, no holes were observed. 4. In the UB treated group III, tissues showed leaching of structural components in the cytoplasm, but no holes were observed. In conclusion, the ultrasonic bath procedure takes approximately 120 minutes from specimen fixation to resin infiltration and gives excellent results.