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 intensity calibration for immunophenotyping by flow cytometry.

R F Vogt1, W E Whitfield, L O Henderson

  • 1National Diabetes Laboratory, Division of Laboratory Sciences, Atlanta, Georgia, 30340, USA.

Methods (San Diego, Calif.)
|June 30, 2000
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

Transcriptional profiling of the L. monocytogenes PrfA regulon identifies six novel putative PrfA-regulated genes.

FEMS microbiology letters·2020
Same author

Nevertheless, She Resisted - Role of the Environment on <i>Listeria monocytogenes</i> Sensitivity to Nisin Treatment in a Laboratory Cheese Model.

Frontiers in microbiology·2020
Same author

Environmental conditions and serotype affect Listeria monocytogenes susceptibility to phage treatment in a laboratory cheese model.

Journal of dairy science·2019
Same author

Performance of succinylacetone assays and their associated proficiency testing outcomes.

Clinical biochemistry·2012
Same author

CFTR mutation analysis and haplotype associations in CF patients.

Molecular genetics and metabolism·2011
Same author

The stability of markers in dried-blood spots for recommended newborn screening disorders in the United States.

Clinical biochemistry·2011

Calibrating fluorescence intensity (FI) using molecules of equivalent soluble fluorochrome (MESF) provides accurate receptor counts. This standardization enhances flow cytometry data comparability across labs and experiments.

Area of Science:

  • Immunology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Flow cytometry commonly uses arbitrary fluorescence intensity (FI) for phenotype classification.
  • Accurate quantification of fluorescent conjugates and stained receptors is challenging with relative FI values.

Purpose of the Study:

  • To establish a standardized method for quantifying fluorescence intensity in flow cytometry.
  • To enable accurate measurement of receptor expression using stoichiometric units.

Main Methods:

  • Calibration of fluorescence intensity (FI) to molecules of equivalent soluble fluorochrome (MESF).
  • Assessment of antibody-binding capacity (ABC) values through binding chemistry analysis.

Main Results:

Related Experiment Videos

  • FI can be expressed in stoichiometric MESF units for precise measurements.
  • Calibration allows for quality control and comparable analysis across different settings.
  • Numeric assessment of ABC values and actual receptor numbers on cells is achievable.

    • Conclusions:

    • Standardized MESF measurements improve the reliability and comparability of flow cytometry data.
    • Accurate receptor quantification is possible through careful calibration and binding chemistry assessment.
    • This approach facilitates robust quality control and inter-laboratory data consistency.