Applicability of a Combined Staining Method with Fluorochromes for the Visualization of Microbial Cells in a Soil Thin Section

概要

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Staining of microbial cells in a soil thin section provides useful information about the role of soil as an ecological niche by relating the in situ distribution of microorganisms to the state of pores under undisturbed conditions. For technical improvement, we developed a procedure consisting of a combined staining method with 5-sulfofluorescein diacetate (SFDA), a metabolic type fluorochrome, and magnesium salt of 1-anilino-8-naphthalene sulfonic acid (Mg-ANS), an adsorption type fluorochrome, in order to detect and differentiate the living microbial cells from the dead ones in a single soil thin section. Using images under a fluorescence microscope, we also examined the applicability of SFDA for staining microbial cells embedded in soil thin sections, which had seldom been attempted. Through this study, we confirmed the following facts: i) combined staining with SFDA and Mg-ANS can be achieved without major modification of the general procedure for thin section preparation, ii) staining with SFDA can be applied to soil thin sections, and iii) detection of in situ distribution and differential identification of the living and dead microbial cells may be possible by the proposed combined staining method.
社団法人日本土壌肥料学会の論文