Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Improved Ultrastructural Preservation of Retinal Cells in Drosophila melanogaster

초고압동결장치를 이용한 초파리 레티나 세포의 향상된 미세구조

  • Mun, Ji-Young (Laboratory of Cell Engineering & 3-D Structure, School of Life Sciences and Biotechnology, Korea University) ;
  • Park, Se-Jin (Laboratory of Cell Engineering & 3-D Structure, School of Life Sciences and Biotechnology, Korea University) ;
  • Han, Sung-Sik (Laboratory of Cell Engineering & 3-D Structure, School of Life Sciences and Biotechnology, Korea University)
  • 문지영 (고려대학교 생명과학대학 생명공학원 세포공학 및 생체3차구조 연구실) ;
  • 박세진 (고려대학교 생명과학대학 생명공학원 세포공학 및 생체3차구조 연구실) ;
  • 한성식 (고려대학교 생명과학대학 생명공학원 세포공학 및 생체3차구조 연구실)
  • Published : 2007.09.30
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Abstract

The Drosophila retinal cell is widely used to study cell development and cell signaling processes. In the past decades, conventional chemical fixation had been used to study the structure of retinal cells in Droscphila. Rapid freezing methods are superior to chemical fixation methods due to their fixation speed. Some Drosophila tissues, such as the eyes, should not be freezed due to their surrounding cuticle layer. Therefore, in the case of the Drosophila retina, the benefits of high pressure freezing and freeze substitution (HPF-FS) had not been verified. In this study, a retinal cell from Drosophila melanogaster had been studied by using the HPF-FS method. Compared to chemical fixation, the preservation of the cytoplasm in the HPF-FS sample was improved on the whole. The HPF-FS cell membranes were smoother than that of chemical fixation. In addition, HPF-FS preserved the mitochondria structures very well. These results of the present study suggest that HPF-FS is superior to other fixation methods for the preservation of the retinal cell structure.

정확한 세포의 구조 분석을 위해서는 조직을 가능한 한 자연 상태 그대로 보존하는 것이 무엇보다 중요하다. 그러나 지금까지 이용되고 있는 화학적인 고정방법은 조직의 변형을 유도하는 것으로 알려져 그 해결방법에 관한 연구가 활발히 진행되고 있다. 그 연구의 결과로 현재 급속 동결법이 제시되었고, 그 중 초고압동결법(high-pressure freezing method)는 가용 두께가 $200{\mu}m$로서 $10{\sim}15{\mu}m$정도인 침윤동결법(plunging method) 혹은 접촉동결법 (slamming method)보다 우수한 방법으로 보고되고 있다. 본 연구팀에는 노랑초파리(Drosophila melanogaster)의 레티나를 화학고정법과 고압동결법으로 고정하여 미세구조를 비교하였다. 먼저 120kV 전자현미경을 이용하여 각 세포 소기관을 비교하였고, 그 중 미토콘드리아의 형태변화를 좀 더 자세히 비교하기 위하여 초고압전자현미경을 이용하였다. 그 결과 급속동결 세포의 세포막과 미토콘드리아의 고정에서 특히 차이가 있음을 알 수 있었고, 이는 주로 탈수에 의한 구조변형 때문인 것으로 추측된다.

Keywords

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