CD4/CD8 IQA Proficiency Testing Program:

Lymphocyte Subset Immunophenotyping Guidelines:

Switch Study Guidelines:

Switching From Two-Platform to Single-Platform Immunophenotyping:

How to Analyze Stabilized Samples:

Stabilized whole blood samples have light scatter and fluorescence staining properties that are different from fresh blood specimens, and as such may not satisfy the automatic gating algorithm defined by the instrument software.  The instrument software may flag the analysis with an error identifying questionable sample integrity. In this instance, manual gating should be applied to the analysis. The gating of the populations should be reviewed to ensure that the population clusters are gated to include the entire cluster without any interfering background and debris events.

BD MultiSET QC Messages for Reference:

Code 3: Sample integrity questionable. Visually inspect.

Code 7: Manual Gate is in effect

For more information, please reference the attached powerpoint, which references gating in BD MultiSET software.  If your team uses a different platform or program, many of the same concepts will still apply.  

Manual Gating of Lymphocytes Using MultiSET