一种基于光密度的适应性微板测试,用于表征动因菌菌体感染
Elijah I Christenson1, Qingyang Zhang2, Ruth Plymale3
1Department of Biology, Ouachita Baptist University.
Journal of visualized experiments : JoVE
|July 17, 2023
概括
这项研究提出了一种新的微板方法来追踪细菌生长和菌体相互作用超过96小时. 开发的协议量化了菌体感染指标,用于缓慢生长的细菌,如actinomycetes.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物信息学是一种生物信息学.
背景情况:
- 菌体 (菌体) 是关键的微生物捕食者,影响自然生态系统中的细菌种群动态.
- 了解菌-细菌相互作用至关重要,特别是在生长缓慢的宿主,如菌体,但需要可靠的长期量化方法.
- 现有的光谱光度法面临着小体积和延长化期的挑战.
研究的目的:
- 开发和验证一种简单,可靠的方法来量化长期的细菌生长在菌体的存在.
- 为了适应标准的96井微板,在长时间 (96小时) 的时间内共同培养菌体和缓慢生长的细菌.
- 建立一个使用光谱光度数据分析菌体感染指标的协议.
主要方法:
- 调整了一个96孔的微板格式,用于在96小时内共同培养细菌和菌体,而无需分样.
- 使用光谱摄影学吸收度测量 (光密度) 每8小时记录一次,以监测细菌生长.
- 使用的R软件用于分析光密度数据以计算感染指标.
- 纳入修改以减轻微板测试中的蒸发和盖子凝结.
主要成果:
- 成功量化了长期的细菌生长和菌体相互作用在96井微板格式.
- 生成的感染指标包括百分比增长抑制,相对毒性和Stacy-Ceballos指数.
- 证明了长期增长曲线实验适应方案的有效性.
结论:
- 概述的程序为进行和分析涉及菌体和生长缓慢的细菌的长期微板生长曲线实验提供了一种有效和强大的方法.
- 这些协议促进了基于微板的测试,这对于研究环境和临床微生物学中的菌体-细菌相互作用至关重要.
- 该方法解决了与长期化小体积相关的技术挑战,提高了高吞吐量能力.
更多相关视频
相关概念视频
Microbial Growth Measurement: Direct Methods
Direct methods for measuring microbial populations in a culture are essential tools in microbiology, providing quantitative data for various applications. Among these, microscopic counts, plate counts, and serial dilution are widely used techniques, each with unique principles and applications.Microscopic CountsMicroscopic counting involves the use of a Petroff-Hausser chamber, a specialized microscope slide with a grid and defined depth. By observing a liquid culture under a microscope,...
Microbial Growth Measurement: Indirect Methods
Estimating microbial growth is essential for understanding population dynamics and environmental adaptations. Indirect methods provide valuable insights by measuring parameters such as turbidity, metabolic activity, and biomass, enabling efficient and reproducible assessments.During exponential growth, microbial cells scatter light proportionally to their biomass, a principle used in turbidity measurements. About one million cells per milliliter produce detectable scattering, which a...
Methods to Assess Microbial Populations
Assessing microbial populations is crucial for understanding microbial roles in health, ecology, and industry. Various complementary techniques—both culture-based and molecular—enable detailed analysis of microbial abundance, diversity, and function.Viable Plate CountThe viable plate count is a traditional culture-based method used to estimate the number of living microbes in a sample. After serial dilution, the sample is spread onto nutrient agar plates. Each viable cell forms a visible...
Microbial Biosensors
Microbial biosensors are analytical devices that utilize living microbes to detect specific substances through measurable signals. These devices consist of two main components: biosensing organisms and signal-transducing elements. Biosensing organisms, such as Escherichia coli or Saccharomyces cerevisiae, are typically housed in multiwell plates connected to transducers, enabling rapid, real-time detection of target analytes.Signal Generation MechanismWhen a target analyte—such as...
Automated Microbial Diagnostics
Automated diagnostic analyzers have transformed clinical microbiology by providing rapid and reliable methods for pathogen identification and antibiotic susceptibility testing. Among these systems, the Vitek 2 is widely used because it automates the traditionally labor-intensive processes of microbial identification (ID) and antibiotic susceptibility testing (AST), delivering standardized and timely results that are essential for effective patient care.Microbial Identification with ID CardsThe...


