Professional Documents
Culture Documents
Immunodiagnostics
A V Differential Technology
and Case Studies.
C
Centrifugation
Disease Management
Hematology
Hemostasis
Lab Automation
Data Management
Flow Cytometry
Primary Care
Bulletin 9151
TABLE OF CONTENTS
DIFFERENTIAL TECHNOLOGY . . . . . . . . . . . . .
Introduction . . . . . . . . . . . . . . . . . . . . .
Differential Methodology: Sample Preparation . . . .
Differential Methodology: Sample Flow and Technology
Flow Cell . . . . . . . . . . . . . . . . . . . .
Dual Focused Flow . . . . . . . . . . . . . . .
Focused Flow Impedance . . . . . . . . . . . .
Differential Methodology: Parameter Derivation . . .
Signal Processing . . . . . . . . . . . . . . . .
DiffPlot Characteristics . . . . . . . . . . . . . .
Lymphocytes . . . . . . . . . . . . . . . . . .
Monocytes . . . . . . . . . . . . . . . . . . .
Neutrophils . . . . . . . . . . . . . . . . . . .
Eosinphils . . . . . . . . . . . . . . . . . . .
WBC/BASO Methodology . . . . . . . . . . . . .
DIFFERENTIAL REAGENT SYSTEM OVERVIEW
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CASE STUDIES . . . . . . . . . . . . . . . . . . . . . . . . . . .
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TABLE
FIGURES
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INTRODUCTION
Volume Aperture
Differential Methodology:
Sample Preparation
Technology
Measurement
Absorbance
cytochemistry and
Volume (ACV
Technology)
Light Absorbance
of Cytochemically
stained cells
Differential Lysis
with Coulter
Principle
Differential
Output
Lymphocyte
Monocyte
Neutrophil
Eosinophil
Basophil
White Cell Count
Whole blood is mixed in the DIFF bath with ACT 5diff FIX reagent incubated
and then stabilized with diluent. This reaction lyses the red cells and stains
the primary granules of monocytes, neutrophils and specific granules of
eosinophils at different intensities. This reaction also preserves the
leukocytes in their native size. The lymphocytes, monocytes, neutrophils
and eosinophils each have unique nuclear and morphologic structure,
and hence absorb light differently. The differential staining and unique
cellular structure provide unique light absorbance characteristics. The
absorbance cytochemistry method is based on the well-known reference
methods of Sheenan et al* and Kass.**
Differential Methodology:
Sample Flow and Technology
Flow Cell
In the flow cell, sequential analyses for cell volume by the Coulter Principle
and light absorbance are performed. The flow cell incorporates a 60 m
aperture for cellular volume analysis and an optical window of 42 m for
light absorbance.
Light
Absorbance
Impedance
Focused flow impedance, using the Coulter Principle, is the technology that
provides cell by cell volume information for each cell as it passes through
the aperture. Each time a cell passes through the aperture a change in
electrical resistance is measured. The change in resistance is proportional
to the volume of the cell.
The ACT 5diff analyzer utilizes dual focused flow in conjunction with
cytochemical staining, light absorbance and Coulter Principle to provide
cellular information for the complete WBC differential.
Differential Methodology:
Parameter Derivation
Each stained cell as it passes through the flow cell is exposed to the light
source and a measurement is taken. As the cell passes through the optical
window, light is scattered in all directions, but only the forward light scatter
is detected by the sensors. The system measures the amount of light lost
due to diffraction and absorbance as compared to full transmission when no
cell is present.
The signals collected are converted into voltage pulses and are processed.
The size and shape of the voltage pulses are equivalent to the unique
nuclear and morphologic structure of the cells being analyzed. The light
absorbance analyzes each cell for cellular content (granularity, nuclear
content) after cytochemical staining. These simultaneous measurements
provide the information for the white cell sub populations, lymphocytes,
monocytes, neutrophils, eosinophils.
Signal Processing
The signals from the impedance aperture and from the absorbance flow
cell are correlated by a window of time. The optical pulse must be detected
within a time window of the impedance pulse, otherwise the signal is
discarded.
The output signals from the focused flow impedance and the light
absorbance measurements are combined to define the white cell differential
population clusters. This combined technology provides an efficient and
robust technology for the white cell differential.
Signal processing is displayed in Figure 2. Parameter DiffPlot development
is displayed in Figure 3.
Incident Light
(Tungsten Lamp)
Flowcell
Detector
No Cell
No Absorbance / Scatter
Base Line Signal
Small Cell
Low Absorbance / Scatter
Low Signal
Volume
Volume
Mono
Eos
Neut
Lymph
Absorbance
Debris
Absorbance
Figure 3. Correlation of Signals to DiffPlot Populations
DiffPlot Characteristics
Lymphocytes
Lymphocytes typically being small with regular shape are smaller in volume
and lower in absorbance than the other cells and are positioned in the lower
part of the DiffPlot. Normal Lymphocyte populations typically have a
homogeneous, gaussian volume distribution.
Neutrophils
Monocytes
Monocytes are typically large cells with kidney shaped nucleus and a
granular cytoplasm. These cells will thus neither scatter nor absorb large
amounts of light and thus are positioned to the low end of the absorbance
axis. However due to their size are positioned high on the volume axis.
Very large monocytes will be found in the IMM (Immature cell) region.
Eosinophils
WBC/BASO Methodology
Whole blood is mixed in the WBC/BASO bath with ACT 5diff WBC Lyse
reagent and stabilized at 35C. This reaction lyses the red cells and
specifically differentiates between basophils and other leukocytes. The
resistance of the basophils to the acidic lytic reagent is the basis for the
differentiation of the basophils from the other leukocytes. The sample is then
pulled with a constant vacuum through an aperture. As each cell passes
through the aperture, it generates a pulse proportional to its cellular volume.
This cellular data is then displayed on the WBC/BASO histogram. The total
leukocyte count and the basophil percentage are determined via the Coulter
Principle and specific thresholds. Figure 4 displays the WBC/BASO
histogram and differential thresholds.
BA1
BA2
BA3
BASO
DIFFERENTIAL REAGENT
SYSTEM OVERVIEW
The ACT 5diff system utilizes cytochemical staining and differential lysis and
the technologies of absorbance cytochemistry and the Coulter Principle to
provide the complete five-part differential. These analyses are performed
simultaneously on the ACT 5diff system.
ACT 5diff Fix reagent contains a lysing agent and the vital stain,
Chlorazole Black, which during the reaction time stains the granules of the
monocytes, neutrophils and eosinophils blue and preserves the leukocytes in
their native size. ACT 5diff Fix is an alcoholic reagent, buffered to a pH of
6.9, which gently lyses all red cells. This reaction provides the dilution used
to differentiate the leukocyte subpopulations using the absorbance
cytochemistry technology.
ACT 5diff WBC Lyse reagent is an acidic solution with a pH of 2.4 that
lyses the red cells and specifically differentiates between the basophils and
other leukocytes. This reaction provides the dilution for the total leukocyte
count and basophil percentage analyses, which are obtained by using the
Coulter Principle and using specific volume thresholds.
ACT 5diff Diluent reagent is used to: dilute the whole blood samples,
stabilize cell membranes for accurate enumeration and sizing, provide the
conductive medium, provide the sheath medium for focused flow and rinses
instrument components after analysis.
DIFFERENTIAL ANALYTICAL /
MECHANICAL SYSTEM OVERVIEW
Sequential Dilution System
Rinse
Bath
First Dilution
Bath
DIFF
Bath
RBS/Plt
Bath
WBC/BA
Bath
Initial aspiration of whole blood occurs at the sampling probe. The SDS
system delivers specific volumes of blood to the respective baths (DIFF or
WBC/BASO) where the sampling needle aligns perfectly with the reagent
inputs. Here, the preheated reagents mix homogeneously with the whole
blood sample. The reagent cleans the sampling needle internally and
externally. Each dilution is then mixed, incubated and stabilized at 35C.
Once the reactions have taken place and the dilutions are ready for analysis,
they need to be transported to the analytical areas.
DIFF Bath
For the DIFF bath sample, the stabilized sample is transported to the
measuring bath in the optical bench. This is accomplished using the syringe
assembly for the DIFF bath. The DIFF syringe feeds the DIFF dilution to the
optical bath sensor while at the same time, two additional syringes provide
diluent sheath flow to the flow cell. Once the sensor detects the sample,
the system begins differential measurements. Figure 6 displays the DIFF
sample flow.
Waste Out
Sheath 1
Sample Flow
for Analysis
Sheath 2
Diff Bath
Sample Flow
to Fill Diff
Syringe
Optical Bench
Waste
Electrodes
Aperture
Light
Source
Optical Window
Electrodes
Sample Injector
Sheath Stream 1
10
Sample Stream
Sheath Stream 2
Detector
WBC/BASO Bath
Bath Wall
WBC/BASO
Bath
Differential Calculation
The results obtained from the DIFF analysis in the flow cell and the WBC/
BASO analysis in the WBC/BASO bath provide the information for the
complete 5-part differential results. The WBC/BASO results provide the
basophil percentage and total white cell count. The DIFF results provide the
lymphocyte, monocyte, neutrophil and eosinophil results. To achieve
differential results equaling 100%, the neutrophil, lymphocyte and monocyte
results are corrected for the results obtained for the basophil analysis.
DIFFERENTIAL FLAGGING
OVERVIEW
VOLUME
Myelocytes,
Metamyelocytes,
Imm. Neutrophils
Blasts,
Promyelocytes,
Monocytes
Imm.
Eos,
A granular
Eos,
Atypical
Neutrophils
Eosinophils
Atyp.
Lymphs,
Blasts
ds
an
Neutrophils
Lymphs
Abnormal
Lymph
Debris
NRBC's,
Lyse Resistant RBC,
Plt Aggregates
ABSORBANCE
Figure 9. DiffPlot Normal and Abnormal Cell Populations
Immature Granulocytes
Band Cells
Band cells are typically larger or of similar size to the neutrophils but due to
their low level of cellular complexity they absorb less light. This places the
band cells in the region between the Neutrophils and the Monocytes.
Blasts
Blast cells are generally larger than monocytes and may have similar
absorbance. Large blasts are included in the IMM cell area.
Atypical Lymphocytes
Small blasts will be found between the normal lymphocyte and monocyte
populations. Large lymphocytes, reactive lymphoid forms, stimulated
lymphocytes and blast cells are found in the ATL region.
12
Mono
Eos
NL
Volume
DiffPlot Flags
Neut
Lymph
SL / SL1
DB
Absorbance
DB Debris
Platelet aggregates
RBCs resistant to lysis (stroma)
NRBCs
SL Small Lymphocytes
Small Lymphocytes
Plt Aggregates
NRBCs, RBCs resistant to lysis
NE, LY, MO, EO, ATL & IMM flagged
SL1 Small Lymph 1
Plt Aggregates
NRBCs, RBCs resistant to lysis
Small abnormal lymphs
NE, LY, MO, EO, ATL, IMM
% & # flagged
NL Neut/Lymph
Small Neut w/o granules
Lymps with segmented nucleus
Neuts with weak membranes
NE, LY % & # flagged
Figure 10. DiffPlot Flags
Volume
DiffPlot Flags
UN
UM
Mono
NE
Eos
MN
Neut
Lymph
LN
Debris
Absorbance
MN Mono/Neut
Monos with hyperbasophilic granules
Immature neuts with nonsegmented nucleus
ATL, IMM % & # flagged
MO, NE % & # inherited ( . . . . )
UM
Upper Mono
Large Monos
Myelocytes, Promyelocytes
Large Blasts
NE, MO, IMM % & # flagged
LN Lower Neut
Fragile or aged neutrophils
Plt Aggs, RBCs resistant to lysis
All WBC Diff parameters flagged
UN Upper Neut
Large neutrophils
Imm Grans (Metamyelocytes, myelocytes, promyelocytes)
NE, IMM % & # flagged
NE Neut/Eos
Immature Eos, Agranular Eos
Atypical Neutrophils
IMM % & # flagged
NE, EO % & # inhibited ( . . . . )
Figure 11. DiffPlot Flags
14
Volume
IMM
Mono
Eos
ATL
Neut
Lymph
Debris
Absorbance
ATL
Large Lymphs
Reactive lymphs
Plasma cells
Blasts
IMM Immature cells
Large Monocytes
Promyelocytes, Myelocytes,
Metamyelocytes
Blasts
Figure 12. DiffPlot Flags
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
5.5 L
5.03
15.2
44.5
89.0
30.3
34.2
17.7
103/L
106/L
g/dL
%
fL
pg
g/dL
%
Range (1)
6.0 / 11.0
4.00 / 6.20
11.0 / 18.8
35.0 / 55.0
80.0 / 100.0
26.0 / 34.0
31.0 / 35.0
10.0 / 20.0
PLT
MPV
169
9.1
103/L
fL
150
6.0
/ 400
/ 10.0
NE
LY
MO
EO
BA
53.8
37.4
6.5
1.6
0.7
%
%
%
%
%
Range (1)
50.0 / 80.0
25.0 / 50.0
2.0 / 10.0
0.0 / 5.0
0.0 / 5.0
NE#
LY#
MO#
EO#
BA#
2.77
1.92
0.33
0.08
0.04
103/L
103/L
103/L
103/L
103/L
2.0
1.0
0.1
0.0
0.0
RBC
VOL
/
/
/
/
/
FLAGS
8.0
5.0
1.0
0.4
0.2
PLT
WBC/
BASO
ABS
History
A normal patient.
Commentary
The ACT 5diff differential correlates
Manual Differential
Neutrophils
Bands
Lymphocytes
16
Morphology
54%
1%
39%
Monocytes
5%
Eosinophils
1%
LYMPHOCYTOSIS
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
3.6 L
5.14
13.8
41.4
81.0
26.8
33.3
15.1
103/L
106/L
g/dL
%
fL
pg
g/dL
%
Range (1)
6.0 / 11.0
4.00 / 6.20
11.0 / 18.8
35.0 / 55.0
80.0 / 100.0
26.0 / 34.0
31.0 / 35.0
10.0 / 20.0
PLT
MPV
188
8.7
103/L
fL
150
6.0
/ 400
/ 10.0
NE
LY
MO
EO
BA
31.3 LL
58.1 HH
6.2
4.0
0.4
%
%
%
%
%
Range(1)
50.0 / 80.0
25.0 / 50.0
2.0 / 10.0
0.0 / 5.0
0.0 / 5.0
NE#
LY#
MO#
EO#
BA#
1.13
2.10
0.22
0.14
0.01
103/L
103/L
103/L
103/L
103/L
2.0
1.0
0.1
0.0
0.0
RBC
VOL
/
/
/
/
/
8.0
5.0
1.0
0.4
0.2
FLAGS
Lymphocytosis
Neutropenia
PLT
WBC/
BASO
ABS
History
A 4-year-old female patient admitted
to the emergency room with fever and
Manual Differential
Morphology
Neutrophils
32%
Commentary
Lymphocytes
60%
Monocytes
5%
Eosinophils
1%
Basophils
1%
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
4.5
2.42
8.5
24.8
102
35.0
34.1
12.8
PLT
MPV
312
7.8
L
L
L
L
H
H
103/L
106/L
g/dL
%
fL
pg
g/dL
%
Range (1)
6.0 / 11.0
4.00 / 6.20
11.0 / 18.8
35.0 / 55.0
80.0 / 100.0
26.0 / 34.0
31.0 / 35.0
10.0 / 20.0
103/L
fL
150
6.0
/ 400
/ 10.0
NE
LY
MO
EO
BA
41.5
22.6
12.7
21.9
1.3
LL%
L
HH
HH
NE#
LY#
MO#
EO#
BA#
1.87 L
1.02
0.57
0.99HH
0.06
Range (1)
50.0 / 80.0
%
25.0
%
2.0
%
0.0
%
0.0
/ 50.0
/ 10.0
/ 5.0
/ 5.0
103/L
103/L
103/L
103/L
103/L
/
/
/
/
/
2.0
1.0
0.1
0.0
0.0
8.0
5.0
1.0
0.4
0.2
FLAGS
WBC: IMM
RBC:
Neutropenia
Eosinophilia
Monocytosis
Macrocytes
PLT
RBC
VOL
WBC/
BASO
ABS
History
A 38-year-old female patient with
admitting diagnosis: Renal failure.
Manual Differential
Neutrophils
Morphology
47%
Bands
Lymphocytes
Monocytes
6%
Eosinophils
26%
Basophils
18
1%
19%
1%
Macrocytosis +2
MONOCYTOSIS
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
15.8 HH 103/L
3.41 L 106/L
11.0
g/dL
31.3 L %
92
fL
32.1
pg
35.0
g/dL
12.2
%
PLT
MPV
190
8.2
103/L
fL
Range
6.0 /
4.00 /
11.0 /
35.0 /
80.0 /
26.0 /
31.0 /
10.0 /
150
6.0
11.0
6.20
18.8
55.0
100.0
34.0
35.0
20.0
/ 400
/ 10.0
(1)
NE
LY
MO
EO
BA
64.1
11.1 LL
22.2 HH
1.1
1.5
Range
%
%
%
%
%
(1)
50.0
25.0
2.0
0.0
0.0
/
/
/
/
/
80.0
50.0
10.0
5.0
5.0
FLAGS
WBC:*WBC, IMM
RBC:
PLT:
NE#
LY#
MO#
EO#
BA#
10.1 HH
1.75
3.51 HH
0.17
0.24 H
103/L
103/L
103/L
103/L
103/L
2.0
1.0
0.1
0.0
0.0
/
/
/
/
/
8.0
5.0
1.0
0.4
0.2
Leukocytosis
Lymphopenia
Neutrophilia
NRBCs
Monocytosis
Platelet
Aggregates
PLT
RBC
VOL
WBC/
BASO
ABS
History
A 64-year-old female admitted with
advanced degenerative bone disease.
Commentary
The ACT 5diff DiffPlot displays a
Morphology
Manual Differential
Neutrophils
Bands
65%
5%
Lymphocytes
13%
Monocytes
17%
Monocytes with
increased
granulation
and numerous
vacuoles.
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
16.3
3.11
10.4
30.1
97
33.3
34.5
13.3
HH
L
L
L
PLT
MPV
82
8.7
103/L
106/L
g/dL
%
fL
pg
g/dL
%
Range (1)
6.0 / 11.0
4.00 / 6.20
11.0 / 18.8
35.0 / 55.0
80.0 / 100.0
26.0 / 34.0
31.0 / 35.0
10.0 / 20.0
103/L
fL
150
6.0
/ 400
/ 10.0
NE
LY
MO
EO
BA
91.5 HH
3.0 LL
2.5
2.0
1.0
%
%
%
%
%
Range (1)
50.0 / 80.0
25.0 / 50.0
2.0 / 10.0
0.0 / 5.0
0.0 / 5.0
NE#
LY#
MO#
EO#
BA#
14.9 HH
0.49 LL
0.41
0.33
0.16
103/L
103/L
103/L
103/L
103/L
2.0
1.0
0.1
0.0
0.0
RBC
VOL
/
/
/
/
/
8.0
5.0
1.0
0.4
0.2
FLAGS
WBC:*WBC, IMM
RBC:
PLT:
Leukocytosis
Neutrophilia
Lymphopenia
Myelemia
Immature Cells
Platelet
Aggregates
PLT
WBC/
BASO
ABS
History
A 48-year-old female admitted with
respiratory failure.
Commentary
The ACT 5diff differential results agree
with the findings from the manual
differential. The automated differential
results (91.5% neutrophils) correlate
well with the thirty-three percent (33%)
bands and sixty percent (60%)
neutrophils reported in the manual
differential. The DiffPlot demonstrates
an increased intensity of cells in the
neutrophil and upper neutrophil regions.
This population of cells generates the
IMM flag. The cells associated with
this flag include myleocytes,
metamyleocytes, promyleocytes, large
neutrophils and blasts. The *WBC flag
indicates white cell count interference
with possibly platelet aggregates or
nucleated red cells. Red cell count
hemoglobin and platelet values are
decreased.
Manual Differential
20
Morphology
Neutrophils
60%
Bands
33%
Lymphocytes
7%
NRBC
1%
GRANULOCYTOSIS, MICROCYTOSIS
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
17.8 HH 103/L
5.13
106/L
12.9
g/dL
37.1
%
72
LL fL
25.1 L pg
34.8
g/dL
16.9
%
PLT
MPV
440 H
7.9
103/L
fL
Range (1)
6.0 / 11.0
4.00 / 6.20
11.0 / 18.8
35.0 / 55.0
80.0 / 100.0
26.0 / 34.0
31.0 / 35.0
10.0 / 20.0
150
6.0
/ 400
10.0
NE
LY
MO
EO
BA
86.2 HH
4.8 LL
7.3
1.0
0.7
%
%
%
%
%
Range (1)
50.0 / 80.0
25.0 / 50.0
2.0 / 10.0
0.0 / 5.0
0.0 / 5.0
NE#
LY#
MO#
EO#
BA#
15.3 HH
0.85 LL
1.30 HH
0.18
0.12
103/L
103/L
103/L
103/L
103/L
2.0
1.0
0.1
0.0
0.0
RBC
VOL
/
/
/
/
/
8.0
5.0
1.0
0.4
0.2
FLAGS
WBC:*WBC
RBC:
PLT:
Leukocytosis
Lymphopenia
Neutrophilia
Monocytosis
NRBC
Microcytes
Platelet
aggregates
PLT
WBC/
BASO
ABS
History
A 70-year-old female admitted in
83%
Bands
9%
Lymphocytes
4%
Monocytes
4%
respiratory failure.
Morphology
Commentary
Platelets
increased,
marked
microcytosis.
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
8.6
3.99 L
11.1
33.9 L
85
27.9
32.9
13.9
103/L
106/L
g/dL
%
fL
pg
g/dL
%
Range (1)
6.0 / 11.0
4.00 / 6.20
11.0 / 18.8
35.0 / 55.0
80.0 / 100.0
26.0 / 34.0
31.0 / 35.0
10.0 / 20.0
PLT
MPV
18 RLL
8.4 R
103/L
fL
150
6.0
/ 400
/ 10.0
NE
LY
MO
EO
BA
6.1 RLL
58.0 HH
33.6 RHH
0.5
1.8
%
%
%
%
%
50.0
25.0
2.0
0.0
0.0
/
/
/
/
/
80.0
50.0
10.0
5.0
5.0
NE#
LY#
MO#
EO#
BA#
0.53 RLL
5.01 H
2.90 RHH
0.04
0.16
103/L
103/L
103/L
103/L
103/L
2.0
1.0
0.1
0.0
0.0
/
/
/
/
/
8.0
5.0
1.0
0.4
0.2
Lymphocytosis
Neutropenia
Immature Cells
Atypical Lymp,
Monocytosis
Plt interpretation
impossible
PLT
RBC
VOL
Range (1)
FLAGS
WBC: UM,ATL,IMM
RBC:
PLT:
SCH
WBC/
BASO
ABS
History
A 78-year-old female admitted with
Acute Myelogenous Leukemia.
Commentary
The ACT 5diff DiffPlot demonstrates
an intense population of cells beginning
in the lymphocyte region extending
through the lymphocytes, atypical
lymphocytes, monocyte and upper
monocyte regions. The lack of
separation within this cellular population
generates the WBC flags including UM,
ATL, IMM and diff parameter R flags.
The cells associated with these flags
include large monocytes, myelocytes,
promyleocytes and blasts, which is in
agreement with the manual differential
results. The platelet histogram displays
marked interference from schistocytes,
generating the platelet interpretation
impossible flag. Reassessment of the
results listed by a reference method
is required. Reference method platelet
results were five.
Manual Differential
Neutrophils
14%
Bands
11%
Metamyelocytes
2%
Blasts
7%
Lymphocytes
Atypical Lymphs
Monocytes
22
Morphology
47%
5%
14%
Numerous
schistocytes, +2
anisocytosis,
platelets
decreased and
numerous
smudge cells.
HB S, SICKEL-CELL CRISIS
WBC
RBC
HGB
HCT
MCV
MCH
MCHC
RDW
5.0
2.39
8.3
23.8
100
34.8
34.9
17.8
PLT
MPV
257
9.7
L
LL
LL
LL
H
H
103/L
106/L
g/dL
%
fL
pg
g/dL
%
Range (1)
6.0 / 11.0
4.00 / 6.20
11.0 / 18.8
35.0 / 55.0
80.0 / 100.0
26.0 / 34.0
31.0 / 35.0
10.0 / 20.0
103/L
fL
150
6.0
/ 400
/ 10.0
NE
LY
MO
EO
BA
33.9
59.4
2.2
3.8
0.7
RLL
RHH
R
R
%
%
%
%
%
Range (1)
50.0 / 80.0
25.0 / 50.0
2.0 / 10.0
0.0 / 5.0
0.0 / 5.0
NE#
LY#
MO#
EO#
BA#
1.69
2.96
0.11
0.19
0.03
RL
R
R
R
103/L
103/L
103/L
103/L
103/L
2.0
1.0
0.1
0.0
0.0
8.0
5.0
1.0
0.4
0.2
Lymphocytosis
Neutropenia
NRBCs
Anemia
PLT
RBC
VOL
/
/
/
/
/
FLAGS
WBC:*WBC,SL,SL1
RBC:
PLT:
WBC/
BASO
ABS
Manual Differential
Morphology
Neutrophils
26%
Lymphocytes
72%
Numerous sickle
cells, +3 target
cells, +1 anisocytosis, rare
large platelet.
Eosinophils
1%
Basophils
1%
NRBC
18/100
WBC
History
A 27-year-old female presented to the
emergency room with shortness of
breath and severe joint and abdominal
pain. Patient has a known history of
Hb S, Sickle-Cell disease. Current
diagnosis: Hb S, Sickle-Cell disease,
crisis phase.
Commentary
The ACT 5diff differential DiffPlot
demonstrates a continuous population
of cells beginning in the lower
lymphocyte region. The lack of
separation within this cellular population
generates the SL and SL1 flags. The
cells associated with these flags include
small lymphocytes, nucleated red cells
and red cells resistant to cell lysis.
Additionally, on the WBC/BASO
histogram a population of cells appears
to the left of the lower threshold. This
population of cells generates the *WBC
flag and differential R flags. These flags
are in agreement with the manual
differential results. The corrected white
cell count is 4.2x103/L. The red cell
histogram displays a markedly wide
distribution in agreement with the mean
cell volume and presence of target
cells. The platelet histogram is
unremarkable.
S I M P L I F Y A U T O M AT E I N N O VAT E
Eastern Europe, Middle East, North Africa: Switzerland, Nyon (41) 22 994 0707. Australia, Gladesville (61) 2 9844 6000. Canada, Mississauga (1) 905 819 1234.
China, Beijing (86) 10 6515 6028. Hong Kong (852) 2814 7431, 2814 0481. France, Villepinte (33) 1 49 90 90 00. Germany, Krefeld (49) 2151 33 35.
Italy, Milan (39) 02 953921. Japan, Tokyo (81) 3 5404 8424. Latin America (1) (305) 380 4709. Mexico, Mexico City (525) 575 6805. Netherlands, Mijdrecht (31) 297 230630.
Singapore (65) 6339 3633. South Africa/Sub-Saharan Africa, Johannesburg (27) 11 805 2014. Sweden, Bromma (46) 8 564 85 900. Switzerland, Nyon 0800 850 810.
Taiwan, Taipei (886) 2 2378 3456. Turkey, Istanbul (90) 216 309 1900. UK, High Wycombe (44) 01494 441181. USA, Brea, CA (1) 800 352 3433, (1) 714 993 5321.
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