The Fixation Effects in Immunohistochemistry and Electron Microscopy Using Low Energy of Microwave (LEM) in Human Gastric Adenocarcinoma and HeLa Cell

사람 위선암과 HeLa 세포에 관한 저에너지 마이크로파 고정효과의 조직화학 및 전자현미경적 연구

  • Yang, Seung-Ha (Department of Pathology, College of Medicine, Soonchunhyang University) ;
  • Son, Tae-Ho (Department of If Engineering, College of Engineering, Soonchunhyang University) ;
  • Shin, Kil-Sang (Department of Life Science, College of Natural Science, Soonchunhyang University)
  • 양승하 (순천향대학교 의과대학 병리학교실) ;
  • 손태호 (순천향대학교 공과대학 IT 공학부) ;
  • 신길상 (순천향대학교 자연과학대학 생명과학부)
  • Published : 2001.06.01

Abstract

Human gastric adenocarcinomas are fixated with low energy of microwave (LEM) to study fixation effects in level of ultrastructure and antigenicity of the cancer. For the Ag-Ab reactions , the LEM fixated sdenocarcinomas are incorporated with monoclonal mouse anti-human p53 (IgG2b, kappa) and rabbit anti human cerbB-2. The retrieval of antigenicity are easily recognizable in the LEM fixated sections compared with that of frozen sections which show often diffused colour reactions. And the LEM fixation methods have preserved ultrastructures of the adenocarcinoma, but it was often difficult to maintain constancy in fixation effects. For the constancy, LEM was coupled with low concentration of chemical fixatives, such as glutaraldehyde (<1%) and $OsO_4$ (<0.5%). The results were acceptable, but there are tendencies that the adenocarcinoma requisitioned rather weak microwave energy to come into the optimal fixation effects. Therefore , cultured HeLa cells were fixated with lower energy of microwave than that used to the adenocarcinoma. The ultrastructures of the single HeLa cell have been preserved. The results may imply that a different energy levels of microwave are requisitioned in accordance with kinds of cells and tissues for the optimal fixation effects. It is reported and discussed that the fixation methods of LEM used in this work could be applied routinely to conceal a insufficient diffusion rate of chemical fixatives into some kinds of cancer without compromising the ultrastructures as well as to improve antigenic quality of frozen sections.