High sensitivity analysis of microorganisms in aquatic environment using image analyzer

水环境中微生物生物量的估计可能成为地球碳平衡讨论的瓶颈，因为众所周知，浮游微生物在水域中占主导地位。因此，我们试图使用图像分析来精确地估计这些生物体积。以细菌为模型微生物，在荧光显微镜下测量各种细胞参数。经证实，DAPI对染色体DNA和吖啶橙染色的单链RNA染色清晰(结果将由Plenum出版公司在《微生物时空多样性》中发表)，并用其生长速度和生长阶段解释了细胞大小的变化。证实细菌在稳定期变小，其组成的单链RNA发生松散，细胞形态发生纵向变化。我们试图解释细菌在细胞周期中的分布通过平板计数确定的不同的生存能力。平板计数细菌数占总数的百分比在C期和D期分布的细胞数内。I期细胞不容易形成集落(结果已提交)。

Estimate of microbial biomass in aquatic environment could be a bottle neck in discussion of carbon balance on earth, because it becomes to be known generally that planktonic microbes predominate in waters. Thus we tried to estimate these biovolume precisely using image analysis. Bacteria was selected as a model microbes and various cellular parameters were measured under epifluorescence microscopy. It was confirmed that DAPI stains clearly chromosomal DNA and acridine orange stains single strand RNA (result will be published from Plenum Publishing Corp.in "Microbial Diversity in Time and Space").The changes in cellular sizes were explained with their growth rate and growth phase. It was confirmed that bacteria became small in stationary phase loosing their constituting single strand RNA and cellular morphological change occurred longitudinally. We tried to explain the varying viability of bacteria determined by plate counting from bacterial distribution in cell cycle. The percent plate counting bacteria to total was within the number of the cells distributed in C- and D-period. Cells in I-period are not considered to form colony easily (result is submitted).