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Related Experiment Videos

High-throughput flow cytometry: validation in microvolume bioassays.

Sergio Ramirez1, Charity T Aiken, Brett Andrzejewski

  • 1Cancer Research and Treatment Center, Department of Pathology, University of New Mexico Health Sciences Center, 2325 Camino de Salud, Albuquerque, NM 87131, USA.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|April 18, 2003
PubMed
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The HyperCyt automated system significantly increases flow cytometry throughput by 10-fold, offering comparable results to manual analysis. This automated autosampler enables efficient microvolume assays, reducing reagent costs and improving sample handling.

Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Cell Biology

Background:

  • Introduced the HyperCyt automated sample handling system for flow cytometry.
  • HyperCyt aspirates samples from microplates for analysis at high throughput rates.
  • This study compares HyperCyt performance against manual flow cytometry and explores microvolume well applications.

Purpose of the Study:

  • To evaluate the HyperCyt automated system's efficiency and accuracy compared to manual flow cytometric analysis.
  • To demonstrate the system's capability in handling microvolume assays.
  • To assess the impact of HyperCyt on sample throughput and reagent usage.

Main Methods:

  • Analyzed human eosinophils, neutrophils, and peripheral blood mononuclear cells for P-selectin glycoprotein ligand-1 (PSGL-1) expression and immunophenotyping.

Related Experiment Videos

  • Utilized U937 cells to evaluate cell suspension stability in microplate wells.
  • Compared HyperCyt autosampling results with conventional manual flow cytometric measurements.

    • Main Results:

    • HyperCyt achieved high throughput (1.5 s/well) with results linearly correlating to manual measurements (r² = 0.98).
    • Accurate discrimination and quantification of lymphocyte subpopulations were achieved across various analysis rates.
    • Microvolume assays (10 µL) in Terasaki plates maintained cell suspension and yielded reproducible results for bead analysis.

    Conclusions:

    • The HyperCyt system provides a 10-fold increase in sample throughput with comparable analytical results to manual methods.
    • Enables efficient 10-microliter assays, leading to reagent cost savings and use of limited reagents.
    • Maintains uniform cell suspensions for extended periods, enhancing assay reliability.