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Related Experiment Videos

Fluorescence intensity as a quality control parameter in clinical flow cytometry.

C W Caldwell1, J Maggi, L B Henry

  • 1Department of Pathology, University of Missouri School of Medicine, Columbia 65212.

American Journal of Clinical Pathology
|October 1, 1987
PubMed
Summary

This study shows that fluorescence intensity and immunostained peripheral blood lymphocyte percentages are useful for monitoring monoclonal antibody (MoAb) quality control and staining procedures in flow cytometry. These parameters aid in identifying issues and evaluating disease states.

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Area of Science:

  • Immunology
  • Biotechnology
  • Clinical Laboratory Science

Background:

  • Quality control (QC) is crucial for reliable flow cytometry results in clinical diagnostics.
  • Standardization of monoclonal antibody (MoAb) staining is essential for accurate peripheral blood lymphocyte (PBL) analysis.
  • Monitoring MoAb performance and staining procedures requires robust QC methods.

Purpose of the Study:

  • To evaluate fluorescence intensity (FI) and percentage of immunostained PBLs as QC parameters for MoAbs and staining.
  • To identify and address variables affecting FI and PBL antigen detection.
  • To establish a standardized flow cytometry QC program for clinical laboratories.

Main Methods:

  • Assessed FI and percentage of immunostained PBLs.

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  • Investigated optimal MoAb dilutions and instrument standardization using fluorescent microspheres.
  • Evaluated day-to-day fluorescence stability with fixed cells.
  • Examined effects of sample separation and storage on FI and PBL antigens.

    • Main Results:

    • While some statistically significant minor changes were observed, FI and MoAb-positive PBL percentages generally proved useful.
    • Day-to-day fluorescence detection showed stability.
    • The study identified key variables impacting staining consistency.

    Conclusions:

    • FI and PBL percentages serve as valuable metrics for a standardized flow cytometry QC program.
    • This QC approach facilitates early detection of methodologic and reagent problems.
    • Routine evaluation of disease states associated with abnormal antigen density is enabled.