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Related Experiment Videos

Immunophenotyping using gold or silver nanoparticle-polystyrene bead conjugates with multiple light scatter.

O Siiman1, K Gordon, A Burshteyn

  • 1Advanced Technology, Beckman Coulter, Miami, Florida 33196-2500, USA. olavi.siiman@coulter.com

Cytometry
|November 21, 2000
PubMed
Summary
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New antibody-conjugated nanoparticles enable simultaneous enumeration of CD4 and CD8 lymphocytes in whole blood using light scatter. This method offers a viable alternative to traditional fluorescent markers for cell analysis.

Area of Science:

  • Nanoparticle-based diagnostics
  • Immunophenotyping
  • Flow cytometry

Background:

  • Antibody-conjugated polystyrene (PS) latex beads are used as light scatter shift agents for lymphocyte enumeration.
  • The range of these agents has been expanded to include gold (Au) and silver (Ag) nanoparticle-aminodextran-PS latex bead conjugates.

Purpose of the Study:

  • To develop and evaluate nanoparticle-based antibody conjugates for simultaneous enumeration of CD4+ and CD8+ lymphocytes in whole blood.
  • To assess the utility of light scatter parameters for lymphocyte enumeration using these novel bead probes.

Main Methods:

  • Utilized a modified flow instrument with multiple light scatter detectors (FS, SS, LMALS, UMALS) for simultaneous bead probe measurements.
  • Employed a conventional flow cytometer for simultaneous bead-fluorescent marker experiments.

Related Experiment Videos

  • Tested mixtures of CD4-PS, CD8-Au-PS, CD4-Au-PS, CD8-PS, CD4-PS, CD8-Ag-PS, and CD4-Ag-PS, CD8-PS beads with 633 nm excitation.

    • Main Results:

    • Successfully enumerated mutually exclusive CD4+ and CD8+ lymphocyte populations simultaneously using mixtures of antibody-conjugated nanoparticle beads.
    • Demonstrated similar enumeration capabilities with both gold and silver nanoparticle conjugates.
    • Showcased simultaneous use of bead probes and fluorescent markers (CD4-RD1/CD8-FITC) in whole blood.

    Conclusions:

    • Enumeration of CD4 and CD8 lymphocytes in whole blood using only light scatter parameters from nanoparticle beads showed good agreement with standard fluorescent marker analyses.
    • The study highlights the potential of nanoparticle-based light scatter agents as an alternative to fluorescent markers for lymphocyte subset analysis.