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KMID : 0381920090390010065
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Korean Journal of Microscopy
2009 Volume.39 No. 1 p.65 ~ p.72
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Observations of the Cyanobacteria Synechocystis sp. PCC 6803 using Cryo-Methods and Cryo-SEM
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Lee Eun-Ju
Kim Youn-Joong
Kweon Hee-Seok
Oh Hyun-Woo
Moon Yoon-Jung
Kim Soo-Jin
Jeong Young-Ho
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Abstract
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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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KEYWORD
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Cryo-Methods, Cryo-SEM, Cyanobacteria, Synechocystis sp. PCC 6803
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