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

Establishing a pure lymphocyte gate for subset analysis by flow cytometry

J M Horvatinovich1, S D Sparks, K P Mann

  • 1Duke University Medical Center, Durham, North Carolina, USA.

Cytometry
|June 15, 1996
PubMed
Summary

A new flow cytometry method improves lymphocyte subset analysis efficiency and cost-effectiveness. This technique enhances purity and accuracy for T cells, B cells, and natural killer cells, crucial for clinical diagnostics.

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

The ownership memory self-reference effect shifts recognition criterion but not recognition sensitivity.

Psychological research·2024
Same author

Molecular impact of selective NFKB1 and NFKB2 signaling on DLBCL phenotype.

Oncogene·2017
Same author

Correlation of extended RAS and PIK3CA gene mutation status with outcomes from the phase III AGITG MAX STUDY involving capecitabine alone or in combination with bevacizumab plus or minus mitomycin C in advanced colorectal cancer.

British journal of cancer·2015
Same author

Rising cost of anticancer drugs in Australia.

Internal medicine journal·2014
Same author

What is clinically relevant prolapse? An attempt at defining cutoffs for the clinical assessment of pelvic organ descent.

International urogynecology journal·2014
Same author

MTF of a 210-70-mm fiber-optic output x-ray image intensifier tube.

Applied optics·2010

Area of Science:

  • Clinical immunology
  • Flow cytometry
  • Immunophenotyping

Background:

  • Current lymphocyte subset analysis methods in clinical flow cytometry face challenges with cost-effectiveness, efficiency, and potential gate contamination.
  • Existing two-color and three-color methods have limitations in excluding non-lymphoid cell populations.
  • There is a continuing need for improved methods for accurate and reliable lymphocyte subset analysis.

Purpose of the Study:

  • To develop a more cost-effective and efficient method for lymphocyte subset analysis.
  • To establish a robust flow cytometry technique for direct measurement of T cell subsets, B cells, and natural killer cells.
  • To improve the purity and accuracy of lymphocyte gating compared to established methods.

Main Methods:

Related Experiment Videos

  • Development of a two-tube staining approach using a mixture of five monoclonal antibodies conjugated to three fluorochromes (FITC, PE, PerCP).
  • Antibody combinations include CD4+CD19, CD3+CD33, CD45, CD8+CD16.
  • Analysis performed on a single-laser flow cytometer.

    • Main Results:

    • The new method demonstrates equivalent recovery and improved purity of the lymphocyte gate compared to established techniques.
    • Clear separation of lymphocytes from other leukocytes and debris is achieved.
    • Accurate separation of T helper/suppressor subsets, natural killer cells, and B lymphocytes is possible.
    • Preliminary data suggests accurate analysis can be performed in a single tube with five antibodies.

    Conclusions:

    • The developed two-tube flow cytometry method offers a cost-effective and efficient approach for lymphocyte subset analysis.
    • This technique provides improved purity and accuracy for immunophenotyping of key immune cell populations.
    • The method facilitates better discrimination of lymphocytes and their subsets, enhancing clinical diagnostic capabilities.