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Related Concept Videos

Microbial Growth Measurement: Direct Methods01:23

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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,...
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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...
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Related Experiment Video

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Colorimetric Detection of Bacteria Using Litmus Test
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A simple colorimetric method for viable bacteria detection based on cell counting Kit-8.

Xianhong Yang1, Yaping Zhong1, Dong Wang1

  • 1Key Laboratory of Textile Fiber and Products, Ministry of Education, Hubei International Scientific and Technological Cooperation Base of Intelligent Textile Materials & Application, Wuhan Textile University, Wuhan 430200, China. ypzhong@wtu.edu.cn.

Analytical Methods : Advancing Methods and Applications
|October 25, 2021
PubMed
Summary

This study introduces Cell Counting Kit-8 (CCK-8) for quantifying live bacteria via a redox reaction. The method offers a rapid, sensitive, and high-throughput approach for bacterial detection.

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Area of Science:

  • Microbiology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Traditional bacterial quantification methods can be time-consuming and labor-intensive.
  • There is a need for rapid and sensitive methods for live bacterial detection.

Purpose of the Study:

  • To introduce and validate the Cell Counting Kit-8 (CCK-8) assay for quantifying live bacterial concentrations.
  • To establish optimal detection conditions and assess the method's performance for common bacterial species.

Main Methods:

  • Utilized the Cell Counting Kit-8 (CCK-8) assay, which relies on the reduction of CCK-8 by bacterial dehydrogenase to formazan.
  • Investigated optimal detection conditions for Staphylococcus aureus and Escherichia coli.
  • Determined linear relationships between absorbance and bacterial concentration (CFU mL⁻¹).

Main Results:

  • Established good linear relationships for S. aureus (R² = 0.9747) and E. coli (R² = 0.9958) over wide concentration ranges.
  • The CCK-8 method achieved detection within 2 hours.
  • Demonstrated consistency with traditional plate counting methods in actual sample analysis.

Conclusions:

  • The CCK-8 based method provides a rapid, sensitive, and efficient approach for viable bacteria detection.
  • This method shows significant potential for high-throughput bacterial quantification.
  • The assay is suitable for analyzing large numbers of samples with high sensitivity.