Interpreting Hematology Scatter-Plots;

O Cancer
One C Center’s
C Keys to S
Seeing the
BIG Picture

Barbara L. Burch, MHA MT (ASCP)

Laboratory Manager
New York University Clinical Cancer Center

Disclosure
• Ms Burch is receiving an honorarium for
her participation in this educational event

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 Learning Objectives
• Give 3 examples of how Complete Blood
Count results are used in the care of
oncology patients.
• Give 2 examples of how practiced Scatter-
Plot interpretation improves oncology
patient care while providing timely and
reliable results to the clinician.
• Identify 2 keys to begin more advanced
hematology Scatter-Plot interpretation.

Current laboratory staffing trends:

1. Decrease in number of accredited schools
2 Decrease in number of students entering
2.
field
3. Bye Bye Baby Boomers
4. Under-recognition of Laboratory
Medicine field inside and outside of
Healthcare
5. Decrease in reimbursement for services
6. Laboratory testing volume is on the rise

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Doing more with less…
• Lab Automation
• LIS Auto-Verification
• POCT
• Cross-Trainingg
• Thorough information
interpretation?

Playing it safe!
• Hold the results – wait for slide
confirmation
• “Catch the false negatives”
• Manufacturer says to
• % or absolute cut off = manual diff
• Blasts counted as Monos…
• Manual WBC’s and PLT’s

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 Routine CBC’s

• Infections
I f ti – viral
i l or bacterial
b t i l
• Anemia – inherited or acquired
(acute or chronic)
• Thrombocytopenia
Th b t i
• Leukemias

CBC’s in Oncology
• Chemotherapy and Radiation Treatments act
b iinterfering
by t f i withith cell
ll division:
di i i
• Neutropenia
• Thrombocytopenia
• Anemia
• Treatments to counteract above side effects:
• Neupogen/Neulasta
• Plt Transfusion
• Pro-Crit/Aranesp, RBC Transfusion

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 Oncology…cont’d
• Recurrence of leukemia
• NRBC’s – Response to growth factor
or tumor invasion of bone marrow?
• Accurate WBC, ANC, Hgb and PLT
results
l needed
d d QUICKLY for f
treatment decisions!!

Fluorescent NRBC

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Removal NRBC influence from WBC and DIFF count
Total WBC Count

WBC NRBC
-
Minus Correct WBC#

NRBC
DIFF - Correct
Minus
Differentials

+ +

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 Platelet
Clumps

Plt
Clumps

Ghost

HPC

IMI Channel
RF
IMI Scattergram
RF

PLT
CLUMPS

Immature Gran
Gh t
Ghost
Ghost
Blasts

HPC

DC

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Fluorescent Optical Plt

RBC Frags

Lg Plts

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Extended Diff – IG Example

Extended Diff – IG Example Cont.

Follow up: Adiff reported as is and smear review

was done to evaluate Blast flag – there were none.

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Using the IMI Channel Example

Using the IMI Channel

Monos

????

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Using the IMI Channel
Patient’s Manual Diff
Neu % = 16
L
Lymphh% = 39
Atyp Lym% = 10
Mono % =0
Myelo % =1
Promeylo % =1
Blasts % = 33
NRBC’s = 5/100 WBC’s
Auer Rods Seen

Pt has untreated AML

Using the IMI Channel - 2

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Using the IMI Channel - 2

Using the IMI Channel - 2

MANUAL DIFFERENTIAL
Segs = 96%
Lymphs = 3%
Monos = 0%
Eos = 1%

Hyper-segmentation of
Neutrophils
(No plt clumps)

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Using the IMI Channel - 2

Hypersegmented Neutrophils
= more nucleic acid = more
fluorescence.

Platelet Clumps - Example

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Plt Clumps – Example continued

Typical IMI plt clump tail

Plt Clumps – Example continued

Smear:

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NRBC’s – Example 1

NRBC’s – Example 1 continued

Uncorrected WBC = 2.0

Treatment decision cutoff = WBC must be > 2.0
Corrected WBC = 1.9 below treatment requirements
Next step – look at ANC

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NRBC’s – Example 2

NRBC’s - Example 2 continued

Instrument gives no reliable

differential information other
than the NRBC’s. WBC
corrected for 30% NRBC’s.
However…

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NRBC’s – Example 2 continued
Patient’s Manual Diff
Neu % = 15
Bands % = 1
Lymph % = 40
Mono % = 1
Atyp Lym% = 43
NRBC’s = 72/100 WBC’s
Micromegakaryocytes = 19/100 WBC’s
Micromegakaryocyte nuclear fragments
15/100 WBC
WBC’ss NRBC count higher on manual
Further WBC correction needed, diff because the instrument is
however – initial correction is done counting the MMKC’s as
by the analyzer so the final WBC’s.
correction is not as drastic.

Blast

Micro-Megakaryocyte

Micro-Megakaryocyte
g y y

Abnormal Platelets

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Plts and RBC Frags Example

Plts and RBC Frags Example

What we were taught to do …
MOAN AND GROAN…
• Rerun and pray for a fitted curve
• Check MCV;; is it low?
• Check MPV; is it high?
• Hurry up and stain the smear and
check under the scope
• If RBC fragments/Microcytes or
Lg Plts present;
•Do manual smear estimate
– OR –
•Do hemacytometer count

What the cancer center does…

• Rerun an optical plt count

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Plts and RBC Frags Example cont.

Plts and RBC Frags Example cont.

Normal Optical Plt Scattergram Patient Optical Plt Scattergram

RBC Fragments

RBC Fragments

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Plts and RBC Frags Example cont.
Smear:

Outcome: Optical plt count verified and released. No smear review done.

Optical Platelet Example

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Optical Platelet Example cont.
Outcome: Optical count reported
without smear follow up, patient
treated.

Clean
S
Separation
i

Just Plain Interesting…

• 66 yr old female – ovarian cancer remission
• Presented with:
• Fevers/chills
• Steadily decreasing Plt count and H&H
• Steadily increasing LDH
• Hematology consult resulted in workup for:
• DIC
• A
Auto-Ab
Ab
• Lyme disease, etc.
• Septicemia
• Babesia/Malarial Infection

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Just Plain Interesting…

Still Just Plain Interesting…

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Just Plain Interesting…

Where are they???

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Compare… Babesia Patient

Time of Dx

Optical Count 86

3 Weeks post
treatment.
PLT-I = 357
PLT-O =351
Optical Count 64

Total WBC Case

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Total WBC Case

Total WBC Case

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Total WBC Case

Total WBC Case Diff Channel Count

WBC/BASO Count

Patient has Multiple Myeloma –

significant rouleaux seen on slide. WBC
smear estimate matched Diff Channel
Count.

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Let’s put it all together…

Let’s put it all together…cont.

IG counted in Diff Channel AND there Re-Ran Optical Plt Count:
is activity in the IMI

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 Let’s put it all together…cont.

Outcome: All results released and Optical plt count without smear
verification.
Smear review done and showed left shift  Myelocytes, 1+ Toxic Gran and
Dohle Bodies.

Keys to Scatter-Plot Interpretation

• UNDERSTAND the technology!
• Know where all the information is
• Know where the flags are generated from
• Look for correlating information in more than
one scatter-plot/histogram
• Develop decision trees
• Look for patterns in scatter-plots
scatter plots and what is
seen on the slide
• What is REALLY important to your
clinicians?

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QUESTIONS???

Barbara Burch email

[email protected]

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