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 Experiment Videos

Fluorescence quenching; a practical problem in flow cytometry.

M R Chapple1, G D Johnson, R S Davidson

  • 1Department of Haematology, West Middlesex University Hospital, Isleworth.

Journal of Microscopy
|September 1, 1990
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Prefrontal High Gamma in ECoG Tags Periodicity of Musical Rhythms in Perception and Imagination.

eNeuro·2020
Same author

Reinventing the disc: a reminder to give credit to past giants.

Proceedings. Biological sciences·2014
Same author

Evaluation of mosquito responses to pyrethroid insecticides topically applied to sheep.

Journal of the American Mosquito Control Association·2013
Same author

Mosquito and West Nile virus surveillance in northeast Montana, U.S.A., 2005 and 2006.

Medical and veterinary entomology·2013
Same author

A forensic investigation into the etiology of bat mortality at a wind farm: barotrauma or traumatic injury?

Veterinary pathology·2012
Same author

Trends in presentation of bone and soft tissue sarcomas over 25 years: little evidence of earlier diagnosis.

Annals of the Royal College of Surgeons of England·2011
Same journal

In operando imaging of the space-charge region in a 4H-SiC MOSCAP using STEM-EBIC.

Journal of microscopy·2026
Same journal

The future of DXA: How AI is transforming bone health diagnostics.

Journal of microscopy·2026
Same journal

The Origins of Ploem's Filter Cube: A Pandora's Box.

Journal of microscopy·2026
Same journal

The reproducibility gap in graph neural network workflows for cell dynamics: A checklist-driven case study.

Journal of microscopy·2026
Same journal

Assessing the reproducibility of a bioimage analysis workflow characterising tissue flow in Drosophila.

Journal of microscopy·2026
Same journal

Modular training resources for bioimage analysis.

Journal of microscopy·2026
See all related articles

R-phycoerythrin (RPE) can quench fluorescein signals in dual fluorescence analysis. This interaction, observed at high RPE concentrations, can affect results from single-laser instruments and necessitates careful control analysis.

Area of Science:

  • Immunology
  • Biotechnology
  • Cell Biology

Background:

  • Dual fluorescence analysis uses fluorochromes with similar excitation and different emission wavelengths.
  • Fluorescein and R-phycoerythrin (RPE) are commonly used dye pairs in single-laser flow cytometry.
  • Previous studies have not reported interactions between fluorescein and RPE.

Purpose of the Study:

  • To investigate potential interactions between fluorescein and RPE in dual fluorescence analysis.
  • To determine the impact of RPE concentration on fluorescein signal intensity.
  • To provide guidance for interpreting dual immunofluorescence data on single-laser instruments.

Main Methods:

  • Dual fluorescence analysis of human tonsil lymphocytes.
  • Labeling cells with monoclonal antibodies conjugated to fluorescein and RPE.

Related Experiment Videos

  • Utilizing a single-laser fluorescence-activated cell sorter.
  • Correlating fluorescein signal reduction with RPE staining intensity.

    • Main Results:

    • High concentrations of RPE were observed to completely quench fluorescein fluorescence.
    • A direct correlation was found between the intensity of red (RPE) staining and the reduction in the fluorescein signal.
    • This quenching effect was demonstrated in dual fluorescence analysis of labeled human lymphocytes.

    Conclusions:

    • RPE can significantly interfere with fluorescein detection in dual fluorescence assays on single-laser flow cytometers.
    • The observed RPE-induced quenching of fluorescein necessitates careful experimental design and data interpretation.
    • Analyzing single-color controls is crucial to mitigate potential misinterpretations caused by this dye-dye interaction.