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Methods to Assess Microbial Populations01:30

Methods to Assess Microbial Populations

100
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...
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Updated: May 3, 2026

Measuring Phagosome pH by Ratiometric Fluorescence Microscopy
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单细胞敏感色度pH检测 基于显微镜比率测量灰度尺度

Shuo Zhou1, Anqi Li1, Linying Zhao1

  • 1School of Environmental and Chemical Engineering, Institute of Carbon Peaking and Carbon Neutralization, Wuyi University, Jiangmen 529020, PR China.

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概括
此摘要是机器生成的。

这项研究引入了一种简单的色度显微镜方法,用于在单细胞水平上精确测量细胞内pH值. 该技术使用红色和蓝色色通道来量化pH值,为生物医学研究提供了一个新的工具.

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科学领域:

  • 生物医学工程 生物医学工程
  • 细胞生物学 细胞生物学
  • 分析化学 分析化学

背景情况:

  • 准确的细胞内pH值测量对于了解细胞过程至关重要.
  • 现有的方法可能很复杂或缺乏单细胞分辨率.
  • 开发可访问的pH监测工具对于生物医学研究至关重要.

研究的目的:

  • 开发和验证一种简单,具有成本效益的方法,用于使用明亮场显微镜在单细胞水平上量化细胞内pH值.
  • 为了建立颜色通道比和细胞内pH值之间的相关性.
  • 为了证明该方法在监测动态pH值变化响应刺激时的适用性.

主要方法:

  • 利用明亮场光学显微镜将细胞图像分成红色 (R) 和蓝色 (B) 颜色通道.
  • 在R和B频道的灰度值上进行了比度运算,以生成比度灰度图像.
  • 通过在不同pH值 (6.00-7.60) 的HeLa细胞与尼日里辛和K+缓冲剂进行化,创建了一个标准曲线.
  • 通过测量A549细胞的细胞内pH值和监测用过氧化刺激HeLa细胞的pH值变化来验证该方法.

主要成果:

  • 确立了pH值与R和B通道的比度灰度之间的强烈相关性,pH值范围为6.00-7.60.
  • 成功测量了A549细胞的细胞内pH值为7.22 ± 0.01.
  • 在过氧化刺激后,在HeLa细胞中对细胞内pH值变化进行了敏感的监测.

结论:

  • 开发的色度显微镜方法提供了一个易于使用的方法,用于在单细胞水平上准确的细胞内pH检测和量化.
  • 这种技术显示出在生物医学中推进与pH相关的单细胞研究的重大前景.
  • 该方法的简单性和有效性使其成为研究研究细胞生理学和疾病的研究人员的宝贵工具.