RUI: Instrumental Methods for the Analysis of Aquatic Bacteria by Flow Cytometry
RUI:流式细胞术分析水生细菌的仪器方法
基本信息
- 批准号:9318432
- 负责人:
- 金额:$ 16.2万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Standard Grant
- 财政年份:1994
- 资助国家:美国
- 起止时间:1994-10-01 至 1997-03-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
9318432 Button Aquatic bacteria control the organic chemistry of natural water systems and comprise about half the biomass of the oceans. Yet, because of recalcitrance to conventional procedures, the identity, phylogenetics, biochemistry and nutritional requirements of most of these organisms, called oligobacteria or ultramicrobacteria (UMB), are unknown. However, UMB cellular properties include extremely small size and DNA content, high affinity for organic nutrients, and resistance to growth in rich media. Flow cytometry offers an ideal window into these organisms not only because of the flexibility and the variety of measurements that can be performed by this instrumentation, but because typical cell numbers in natural populations are ideally suited for this type of direct analysis, natural cell populations can be preserved for analysis at a later time, and the single-cell basis of the analysis can provide a wealth of information on cell population structure. Unfortunately, present flow cytometry analytical methods have been developed only for large cells. Techniques for treatment and standard size curves for UMB-type cells have to be developed. The objective of the present research project is to develop methods for efficient analysis of UMB by flow cytometry. Standard curves for cell size and for DNA per cell will be completed. Additional standard curves will be generated for UMB RNA and protein content. Membrane potential will be used to probe the physiological state of cells, and the feasibility of using oligonucleotide probes as a taxonomic tool for oligobacteria will be examined. The concept of deriving three-dimensional spacing and chemotaxis information about bacteria in mixed systems from signal-proximity analysis will be tested under both laboratory and field conditions. %%% Aquatic bacteria, especially the oligobacteria or the ultramicrobacteria (UMB), comprise almost one half the biological material in the sea and, as such their metabolic activity can control the chemistry of the world's oceans. Surprisingly, however, very little is known about these cells except that they are small, they contain a minimum amount of DNA, and although they have a strong attraction to organic compounds, they only grow when the concentration is very low. The objective of the present research program is to develop methods using flow cytometry to learn about the cells directly in sea water. Standard curves will be developed to measure cell size and estimate their DNA, RNA, and protein contents. Novel methods will be used to determine which cells are metabolizing and how many different UMB-cell types may be present in a water sample. The feasibility will be tested in the laboratory and in the field of analyzing cell movement under flow cytometer conditions to determine if the aquatic bacteria exhibit tactic behavior toward desirable nutrients. %%%
水生细菌控制着天然水系统的有机化学成分,占海洋生物量的一半左右。然而,由于对常规程序的抗拒,大多数这些被称为寡菌或超微生物(UMB)的生物的身份,系统发育,生物化学和营养需求是未知的。然而,UMB的细胞特性包括极小的尺寸和DNA含量,对有机营养物质的高亲和力,以及在富培养基中生长的抗性。流式细胞术为这些生物提供了一个理想的窗口,不仅因为这种仪器可以进行灵活多样的测量,而且因为自然群体中的典型细胞数量非常适合这种类型的直接分析,自然细胞群体可以保存以供以后分析,并且分析的单细胞基础可以提供丰富的细胞群体结构信息。不幸的是,目前的流式细胞术分析方法仅适用于大细胞。必须开发针对mb型细胞的处理技术和标准尺寸曲线。本研究项目的目的是发展流式细胞术对UMB进行有效分析的方法。将完成细胞大小和每个细胞DNA的标准曲线。将生成UMB RNA和蛋白质含量的附加标准曲线。将利用膜电位来探测细胞的生理状态,并探讨利用寡核苷酸探针作为寡细菌分类工具的可行性。从信号接近分析中获得混合系统中细菌的三维间距和趋化性信息的概念将在实验室和现场条件下进行测试。水生细菌,特别是寡菌或超微细菌(UMB),几乎占海洋生物物质的一半,因此它们的代谢活动可以控制世界海洋的化学成分。然而,令人惊讶的是,我们对这些细胞知之甚少,只知道它们很小,含有最少的DNA,尽管它们对有机化合物有很强的吸引力,但它们只有在浓度很低的情况下才能生长。本研究计划的目的是发展使用流式细胞术直接了解海水中的细胞的方法。将开发标准曲线来测量细胞大小并估计其DNA, RNA和蛋白质含量。新的方法将用于确定哪些细胞正在代谢,以及水样中可能存在多少种不同的umb细胞类型。可行性将在实验室进行测试,并在流式细胞仪条件下分析细胞运动,以确定水生细菌是否表现出对所需营养物质的策略行为。%%%
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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{{ truncateString('Don Button', 18)}}的其他基金
Activity and Characteristics of the Dissolved Organic Carbon-Dependent Picoplankton over the Annual Cycle in a Subarctic Lake
亚北极湖泊溶解有机碳依赖的超微型浮游生物年周期的活动和特征
- 批准号:
0243913 - 财政年份:2003
- 资助金额:
$ 16.2万 - 项目类别:
Standard Grant
Activity of Individual Aquatic Bacteria by Flow Cytometry; Instrumentation and Protocols
通过流式细胞术检测单个水生细菌的活性;
- 批准号:
0096748 - 财政年份:2001
- 资助金额:
$ 16.2万 - 项目类别:
Standard Grant
Functionality of Dilute-Environment Bacteria
稀环境细菌的功能
- 批准号:
0085628 - 财政年份:2000
- 资助金额:
$ 16.2万 - 项目类别:
Standard Grant
Dynamic Properties of Picoplankton Over the Annual Cycle in Near-Polar Lakes
近极地湖泊微型浮游生物年度循环的动态特性
- 批准号:
9907776 - 财政年份:1999
- 资助金额:
$ 16.2万 - 项目类别:
Continuing Grant
LEXEN: Characteristics of Bacteria Native to Extremely Dilute Environments
LEXEN:极度稀释环境中原生细菌的特征
- 批准号:
9714298 - 财政年份:1997
- 资助金额:
$ 16.2万 - 项目类别:
Standard Grant
Distribution, Nutrition, Physiology and Phylogenetics of Oligobacteria
寡杆菌的分布、营养、生理学和系统发育学
- 批准号:
9316731 - 财政年份:1994
- 资助金额:
$ 16.2万 - 项目类别:
Continuing Grant
Sensitivity, Distribution, Characterization and Production of Ultramicrobacteria
超微细菌的敏感性、分布、表征和生产
- 批准号:
9019625 - 财政年份:1991
- 资助金额:
$ 16.2万 - 项目类别:
Continuing Grant
REU: Refinement of Instrumentation, Protocols, and Calibrations for the Analysis of Marine Bacteria by Flow Cytometry
REU:流式细胞术分析海洋细菌的仪器、协议和校准的改进
- 批准号:
8822155 - 财政年份:1989
- 资助金额:
$ 16.2万 - 项目类别:
Standard Grant
Development of a Facility for Flow Cytometry
流式细胞术设施的开发
- 批准号:
8518397 - 财政年份:1986
- 资助金额:
$ 16.2万 - 项目类别:
Continuing Grant
Development of a Facility for Flow Cytometry
流式细胞术设施的开发
- 批准号:
8411061 - 财政年份:1984
- 资助金额:
$ 16.2万 - 项目类别:
Standard Grant
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