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Department of Laboratory Medicine University of Washington

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Presentation on theme: "Department of Laboratory Medicine University of Washington"— Presentation transcript:

1 Department of Laboratory Medicine University of Washington
Data Analysis Department of Laboratory Medicine University of Washington

2 Data Analysis Assess data quality Identify populations
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

6 Degeneration Decrease in intensity for many antigens

7 Viability Gate

8 Viability Gate All cells Viable cells

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

15 Example Time

16 Example Singlets

17 Example Viable

18 Determining Positivity

19 Determining Positivity
Incorrect Correct

20 Population Identification

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

22 Cell Type Identification
Borowitz et al (1993) AJCP 100: Steltzer et al (1993) Ann NY Acad Sci 667:

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 Follicular Lymphoma

27 ALL MRD 0.1% abnormal immature B cells

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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