Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

57.1K
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
57.1K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Rapid Identification of Yeasts in Foods Based on Thermal-Scanning Fluorescence Spectroscopy.

Analytical chemistry·2026
Same author

Cell viability assessment by using GelRed/SYTO 9-based double staining.

Methods and applications in fluorescence·2025
Same author

Intrafibrillar mineralization of type I collagen by micelle-loaded amorphous calcium phosphate nanoparticles.

RSC advances·2023
Same author

Bridge-DNA synthesis triggered by an allosteric aptamer for the colorimetric detection of pathogenic bacteria.

Analytical methods : advancing methods and applications·2023
Same author

On-bead DNA synthesis triggered by allosteric probe for detection of carcinoembryonic antigen.

Mikrochimica acta·2022
Same author

Using a safe and effective fixative to improve the immunofluorescence staining of bacteria.

Methods and applications in fluorescence·2021

Related Experiment Video

Updated: Jun 22, 2025

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells
07:23

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

Published on: September 5, 2013

43.8K

Rapid and Sensitive Quantification of Bacterial Viability Using Ratiometric Fluorescence Sensing.

Shengbin He1, Yajing Chen1, Jingtong Wang1

  • 1Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education, Guangxi Colleges and Universities Key Laboratory of Biological Molecular Medicine Research, School of Basic Medical Sciences, Guangxi Medical University, Nanning, Guangxi 530021, P. R. China.

Analytical Chemistry
|June 27, 2024
PubMed
Summary
This summary is machine-generated.

A new ratiometric fluorescent method using GelRed and SYTO 9 effectively distinguishes live from dead bacteria. This sensitive technique aids in food safety and drug development by accurately assessing bacterial viability.

More Related Videos

Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens
10:39

Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens

Published on: June 12, 2013

13.5K
Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer
08:27

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

9.1K

Related Experiment Videos

Last Updated: Jun 22, 2025

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells
07:23

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

Published on: September 5, 2013

43.8K
Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens
10:39

Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens

Published on: June 12, 2013

13.5K
Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer
08:27

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

9.1K

Area of Science:

  • Microbiology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Bacterial viability assessment is crucial for food safety and antimicrobial research.
  • Traditional methods for viability testing can be limited in sensitivity and accuracy.

Purpose of the Study:

  • To develop a novel, sensitive, and accurate ratiometric fluorescent method for bacterial viability assessment.
  • To establish a method for distinguishing between live and dead bacteria using DNA-binding dyes.

Main Methods:

  • Utilized GelRed as a DNA-binding stain, noting live bacteria exclude it while dead bacteria absorb it.
  • Employed cations to minimize nonspecific adsorption of GelRed.
  • Developed a double-staining technique with SYTO 9 (a membrane-permeable dye).
  • Established a ratiometric fluorescent assay for quantifying dead bacteria.

Main Results:

  • The GelRed/SYTO 9 method demonstrated high sensitivity, detecting dead bacteria at levels as low as 0.1% in Escherichia coli O157:H7.
  • A strong linear correlation (R² = 0.97) was observed between ratiometric fluorescence and the theoretical dead bacterial ratio.
  • The method proved effective in evaluating the germicidal efficacy of heat treatments, showing bacterial death at 50 °C.

Conclusions:

  • The developed ratiometric fluorescent method offers superior sensitivity, accuracy, and safety compared to traditional viability assays.
  • This technique has broad applications in microbial research, pathogen detection, and drug development.
  • The method shows potential for adaptation to mammalian cell viability studies.