Observations of the Cyanobacteria Synechocystis sp. PCC 6803 using Cryo-Methods and Cryo-SEM

Cryo-Methods와 Cryo-SEM을 이용한 Cyanobacteria, Synechocystis sp. PCC 6803 미세구조 관찰

  • Lee, Eun-Ju (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Moon, Yoon-Jung (Proteome Research Team, Korea Basic Science Institute) ;
  • Oh, Hyun-Woo (Industrial Bio-materials Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Su-Jin (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Chung, Young-Ho (Proteome Research Team, Korea Basic Science Institute) ;
  • Kweon, Hee-Seok (Division of Electron Microscopic Research, Korea Basic Science Institute) ;
  • Kim, Youn-Joong (Division of Electron Microscopic Research, Korea Basic Science Institute)
  • 이은주 (한국기초과학지원연구원 전자현미경연구부) ;
  • 문윤정 (한국기초과학지원연구원 프로테옴연구부) ;
  • 오현우 (한국생명공학연구원 산업바이오소재 연구센터) ;
  • 김수진 (한국기초과학지원연구원 전자현미경연구부) ;
  • 정영호 (한국기초과학지원연구원 프로테옴연구부) ;
  • 권희석 (한국기초과학지원연구원 전자현미경연구부) ;
  • 김윤중 (한국기초과학지원연구원 전자현미경연구부)
  • Published : 2009.03.31

Abstract

Cryo-SEM which enables specimens to be observed in frozen form has been used to study liquid specimens in their native states. Cryo-methods, sample preparation for cryo-SEM, are quite complex and involve several discrete but vitally interconnected steps which are rapid cooling, fracturing, sectioning, etching and coating. It is important to select practical techniques and to optimize conditions of each steps considering analytic purpose and specimen characters, viz., sample dimension, water contents. In this study, etching methods and sample preparation before freezing had been studied for observation of cyanobacteria, Synechocystis sp. PCC 6803 using cryo-SEM and their cryo-SEM images were compared to Conventional SEM (CSEM) images treated by chemical fixation. We could observe the improved morphological images of the pili of the surface and membranes of Synechocystis sp. PCC 6803 and the three-dimensional architectures of their biofilm, which were difficult to observe using chemical fixation and CSEM. These results suggest that cryo-methods/cryo-SEM are useful techniques for morphological study of biological specimen.

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