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Data Analysis

Data Analysis. Department of Laboratory Medicine University of Washington. Data Analysis. Assess data quality Remove artifacts Identify populations Compare with normal Identify abnormal populations Quantitate and evaluate immunophenotype Generate report. Assess Data Quality.

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Data Analysis

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  1. Data Analysis Department of Laboratory Medicine University of Washington

  2. Data Analysis • Assess data quality • Remove artifacts • Identify populations • Compare with normal • Identify abnormal populations • Quantitate and evaluate immunophenotype • Generate report

  3. Assess Data Quality

  4. Detector Optimization Negative populations entirely on scale

  5. Degeneration Increase SS Decrease FS 08-03307

  6. Degeneration Decrease in intensity for many antigens 08-03307

  7. Viability Gate 08-03307

  8. Viability Gate All cells Viable cells 08-03307

  9. Sample Exhaustion Air in system gives rise to many spurious signals Event gate to exclude non-real events

  10. Laser Delay Fluidic instability - Monitor events over time to detect

  11. Laser Delay Original Gated

  12. Doublet Discrimination

  13. Doublet Discrimination • Doublets = > one cell in laser simultaneously • High cell concentrations • Cell aggregates, sample preparation • High sample aspiration pressure • Doublets have composite properties • Can exclude using height, area, or width

  14. Example Original 07-04513

  15. Example Time 07-04513

  16. Example Singlets 07-04513

  17. Example Viable 07-04513

  18. Determining Positivity

  19. Determining Positivity Incorrect Correct 07-08661

  20. Population Identification

  21. Cell Type Identification Lymphocyte population identified by FS/SS gating

  22. Cell Type Identification Borowitz et al (1993) AJCP 100:534-40. Steltzer et al (1993) Ann NY Acad Sci 667:265-280

  23. Lineage Identification • CD19 for B cells and CD3 for T cells • Assumptions that may not always be correct • Always use at least two methods of identification

  24. Compare with Normal

  25. Normal B cell Maturation Wood and Borowitz (2006) Henry’s Laboratory Medicine

  26. Follicle Center B cells Follicular Hyperplasia 08-03324 Follicular Lymphoma 08-01359

  27. ALL MRD 0.1% abnormal immature B cells 06-01469

  28. Data Analysis • Data displayed as dot plots or histograms • Restrict to subset having high informational content • Color discrete populations • Display information from other parameters • Allow rapid visual identification in multiple plots • Display data in consistent manner • Pattern recognition

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