SEM observation of Fresh Water Microorganisms Prepared by the Water Freeze-drying Method - 2 (Protists)

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  • 水凍結乾燥法を用いた淡水産微小生物のSEM試料作製と観察－2（原生生物）
  • ミズ トウケツ カンソウホウ オ モチイタ タンスイサン ビショウ セイブツ ノ SEM シリョウ サクセイ ト カンサツ(2)ゲンセイ セイブツ

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Abstract

<p>　For the SEM observation of protists, the specimens are generally fixed chemically in advance. The fixatives, however, are toxic, and the solution used must be processed as hazardous waste. Since the optimum fixatives and the fixing conditions differ depending on the species, a large amount of time may have to be wasted for finding the optimum condition. Fixation is a major obstacle to the use of SEM. Freshwater protists were prepared for SEM by using the water freeze-drying method without using any chemicals, and its morphological damage was investigated. The observations of protists collected from lakes, marshes, and rice fields in the Kanto area revealed that the shapes and microstructures, e.g. cilia and flagella, were well preserved. With this method, the identification accuracy of microorganisms, which was difficult to identify by optical microscopy, was improved. If fixation is not required, various species can be processed together. Plankton survey using SEM was not practical because it required a considerable amount of work and was time consuming. We applied this method to the survey of protists in the surface waters of Lake Fujigoko. The survey results obtained by the SEM method are almost the same as those obtained using the optical microscope. In addition, the possibility of further improvement of the detection rate was also clarified. Since the SEM specimen is dehydrated, long-term preservation is possible by storing it in a desiccator. It is a great advantage of this method that it is possible to make verifications of the survey result at a later time.</p>

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