Interfering Sub in Hematology

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EXIT
Introduction HGB Interferences
To navigate
Lipemia
through this WBC Interferences
document, Plasma Replacement Procedure
Fragile WBCs
click on an Plasma Hemoglobin Procedure
Clumped WBCs
underlined Indirect Estimation Procedure
WBC Count Overrange
link to go to
the indicated WBC Estimation Using Peripheral Smear Platelet Interferences
section. Pseudothrombocytopenia—Spurious Platelet Clumping
RBC Interferences
Platelet Satellitism
Hemolyzed Specimens
Giant Platelets
Spuriously Decreased Hematocrits
Megakaryocytes
Cold Agglutinins
Falsely Elevated/Overrange Platelet Count
Lyse-resistant RBCs
Alternative Platelet Procedures
Hyperglycemia
Sodium Citrate Procedure for Platelet Clumping
Platelet Estimate Using Peripheral Smear
Manual Platelet Count
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Interfering Substances in Hematology Resource Book CELL-DYN® Systems 69927-101 November 2001
WBC Interferences RBC Interferences HGB Interferences Platelet Interferences
MENU INTRODUCTION
BACK
Many specimen conditions can cause changes in the scattergrams, histograms, and other results
NEXT
generated by Hematology analyzers. The following may be considered substances that interfere
EXIT with Hematology results:
• Specimens from patients with diseases such as leukemia, autoimmune or inflammatory disorders,
and neoplasias, produce unusual scattergrams and WBC counts.
• Abnormal RBC populations, diseases such as anemia, or less than ideal specimen collection can
alter histograms and RBC counts.
• Hyperlipidemia can affect scattergrams and also the hemoglobin reading.
• Platelet abnormalities due to size discrepancies, specimen collection, or disease states can alter
scattergrams, histograms and platelet counts.
For your use as a reference tool in your laboratory, Abbott Diagnostics is offering a discussion of
many of these substances with some suggestions for minimizing the interference.
For many interfering substances, knowing what to look for will make dealing with them easier. You
will get the results to the physicians faster and with more confidence. To make this Resource Book
easy to follow, Abbott Diagnostics has put the topic headings across the top in a color-coded format.
page 2
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page 3
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MENU WBC INTERFERENCES

BACK
Fragile WBCs
NEXT
EXIT Definition
Fragile WBCs are easily fragmented, smudged, or destroyed. These cells may occur in specimens
from patients with acute or chronic lymphocytic leukemia (CLL). In CLL, the fragile cells are a
phenomenon associated with the disease. Fragile cells may also be present in specimens from
patients who are taking immunosuppressive drugs or who have renal disease. Fragile WBCs may
be either lymphoblasts (in ALL) or lymphocytes (in CLL) or neutrophils (in patients receiving
chemotherapy or anti-inflammatory drugs).
Impact of Interference
On the CELL-DYN 3000/3500/3700 Systems, fragile cells are detected by monitoring the count rate
for the duration of the WBC count. A summary of the count rate is accessed from the Diagnostics
menu by pressing COUNT RATE SUMMARY, then WBC (WOC) COUNT RATE. This summary
shows the data obtained during the count (time, count, and rate) at 0.5 second intervals for 7.5
seconds. In the case of an extended count, this information is also given for an additional 7.5 second
interval. The data summary can be displayed as a graph by pressing WBC (WOC) RATE GRAPH.
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MENU If a 10% or greater decline in count rate is detected, an FWBC flag is displayed, indicating the
presence of fragile WBCs.
BACK
Corrective Actions
NEXT
• Examine a stained smear for the presence of “smudge cells.”
EXIT
• Prepare a smear with albumin according to your laboratory protocol if desired.
• If the WBC flag is displayed, verify the WBC count using the WBC Estimation Using Peripheral
Smear procedure contained in this document.
• If the DFLT or DIFF flag is displayed, verify the WBC differential according to your laboratory
protocol using a peripheral smear.
References
CELL-DYN 3500 System Operator’s Manual, 92722-01, 3–55.
Luke RA, Koepke JA, Siegel RR. The effects of immunosuppressive drugs and uremia on automated
leukocyte counts. American Journal of Clinical Pathology. 1971;56:503–507.
page 5
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SUSPECT
MENU
WBC 82.7 K /µL (WIC)
BACK NEU 1.83 2.22 %N
LYM 79.4 96.0 %L VAR LYM
NEXT MONO .381 .460 %M FWBC
EOS .070 .084 %E
EXIT
BASO 1.05 1.26 %B DFLT (NLMEB)
RBC 4.09 M /µL

HGB 13.2 g/dL
HCT 38.0 %
MCV 93.1 fL
MCH 32.3 pg
MCHC 34.7 g/dL
RDW 15.4 %
PLT 125 K /µL
Sequence 2117: Fragile WBCs
page 6
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MENU
BACK
NEXT
EXIT
Smudge Cell
page 7
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DIAGNOSTICS MENU
MENU Ready Operator ID
Sequence # 2117
BACK
7675.7
NEXT
6716.3
EXIT
5756.8
4797.3
3837.9
2878.4
1918.9
959.5
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5
WOC CNT GRAPH
page 8
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MENU Clumped WBCs

BACK Definition
NEXT The presence of clumped (aggregated) WBCs can be due to a variety of causes that may change the
EXIT chemical makeup of the WBC’s cell membrane. A pathological condition, such as an autoimmune
disease or inflammation, can produce this effect. Clumped WBCs can also be the result of chemical
agents, such as chemotherapeutic drugs or drugs used to treat renal disease.
Clumped WBCs result in a falsely decreased WBC count. WBC clumps may be observed along the
top right and far right sides of the 0˚/10˚ scatterplot, as well as other scatterplots such as the 90˚/10˚
scatterplot and mono-poly (M-P) histogram.
In addition, if the clumped WBCs disassociate during the counting cycle, a WOC flow error may
occur. This is related to the instrument’s evaluation of the count rate data (see Fragile WBCs).
However, in this case, count rate increases, rather than declines.
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MENU Corrective Actions

BACK • Review the 0˚/10˚ scatterplot, the 90˚/10˚ scatterplot, and the M-P histogram for evidence of
WBC clumps.
NEXT
• If a WOC flow error occurs, review the WBC count rate summary graph for an inclining count
EXIT rate.
• Follow your laboratory protocol and examine a stained smear and verify the WBC count by using
an alternative method.
Reference
Bizzaro, N. Granulocyte aggregation is acetic acid and temperature dependent. Archives of Pathology
and Laboratory Medicine. 1993;117:528–530.
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SUSPECT
MENU
WBC 10.6 K /µL
BACK NEU 8.94 84.3 %N IG/BAND
LYM .423 3.99 %L
NEXT MONO 1.19 11.2 %M
EOS .034 .316 %E
EXIT
BASO .019 .181 %B
RBC 3.63 M /µL

HGB 11.1 g/dL
HCT 33.0 %
MCV 90.9 fL
MCH 30.5 pg
MCHC 33.5 g/dL
RDW 15.2 %
PLT 201. K /µL

MPV 8.30 fL
Sequence 6858: Clumped WBCs
page 11
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MENU
BACK
NEXT
EXIT
Clumped WBCs
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MENU WBC Count Overrange

BACK Definition
NEXT In this condition, the WBC count exceeds the linearity limits of the instrument. An extremely
EXIT elevated WBC count is most commonly associated with leukemia or other myeloproliferative disor-
ders. Exceptions may be seen in leukemoid reactions such as acute infections, severe metabolic
inflammatory reactions, or neoplastic processes.
Chevrons (>>>>) appear in place of the WBC count (WOC greater than 250 K/µL for CELL-DYN
3500/3700; WBC greater than 99.9 K/µL for CELL-DYN 3000). The data log from the CELL-DYN
3500/3700 System displays the actual WIC and WOC values. These values may be used to help
determine the dilution necessary to bring the WBC count within the linearity limits of the instrument.
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BACK • Dilute a well-mixed aliquot of whole blood with diluent.
NEXT Examples:
0.5 mL whole blood + 0.5 mL diluent (1:2 dilution) → multiply WBC by 2
EXIT
• Run the well-mixed dilution as a patient.
NOTE: The specimen may be run in the Auxiliary Mode on the CELL-DYN 3500/3700.
• For specimens run on the CELL-DYN 3000, multiply the WBC result by the dilution factor before
reporting. The Auxiliary Mode on the CELL-DYN 3500/3700 automatically corrects the WBC
count for the dilution factor.
References
CELL-DYN 3500 System Operator’s Manual 92722-01.
Harmening D, ed. Clinical Hematology and Fundamentals of Hemostasis. Philadelphia, PA: F.A.
Davis; 1992:243.
page 14
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MENU SUSPECT
WBC >>>> K/µL
BACK
NEU 4.39 % N (WOC) WBC
NEXT LYM 65.7 % L BLAST
MONO 13.7 %M DFLT (NLMEB)
EXIT EOS .102 %E
BASO 16.1 %B
RBC 3.21 M /µL

HGB 11.0 g/dL
HCT 26.2 %
MCV 81.4 fL RBC MORPH
MCH 34.1 pg
MCHC 41.9 g/dL
RDW 16.4 %
PLT 64.8 K /µL

MPV fL LURI
Sequence 1394: WBC Count Overrange
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SUSPECT
MENU
WBC 75.8 K /µL
BACK NEU 1.83 2.41 %N BANDs
LYM 66.5 87.7 %L
EOS .046 .060 %E
EXIT
BASO 1.18 1.56 %B DFLT (NE)
RBC .355 M /µL

HGB 1.28 g/dL
HCT %
MCV fL
MCH pg
MCHC g/dL SAMPLING ERR
RDW %
PLT 4.21 K /µL

MPV fL
Sequence 1397: 1:10 Dilution of Sequence 1394
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MENU WBC Estimation Using Peripheral Smear

BACK Purpose
NEXT Two forms of this procedure (A and B) are used to estimate the total white blood cell count by micro-
EXIT scopically examining a stained smear. Both forms use the same specimen, but vary in specific steps.
Specimen
Wedge smear made from fresh EDTA-anticoagulated blood producing an even, feathered edge. Stain
the smear using laboratory protocol.
Procedure A
1. Scan the smear under a 10x (low) objective to ensure even distribution of WBCs.
2. Examine the smear under a 40x (high dry) objective. Select an area of the smear where the RBCs
are just touching each other and count all the WBCs, including ruptured cells, in 10 successive
fields.
3. Divide the total number of WBCs counted by 10 to get the average number of WBCs per high dry
field.
4. Multiply the average number of WBCs per high dry field by two to get the WBC estimate.
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MENU Calculation
BACK
WBC estimate = # of WBCs counted in 10 fields x 2
NEXT 10
EXIT Procedure B
1. Follow steps 1–3 in Procedure A above. Do not truncate the number.
2. Using Table 1 on the following pages, obtain the estimated WBC count as follows:
a. Find the whole number in the column and the decimal value in the top row.
b. The number at the intersection of these two points is the estimated WBC count.
Example:
If average WBCs/40x (high dry) field = 5.2, then the WBC estimate is 10.4.
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MENU Table 1: WBC Estimate Chart (40x High)

BACK 1 WBC = 2.0 x 103/µL (continued)
.0 .1 .2 .3 .4 .5 .6 .7 .8 .9
NEXT
0 0.0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8
EXIT
1 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8
2 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8
3 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8
4 8 8.2 8.4 8.6 8.8 9 9.2 9.4 9.6 9.8
5 10 10.2 10.4 10.6 10.8 11 11.2 11.4 11.6 11.8
6 12 12.2 12.4 12.6 12.8 13 13.2 13.4 13.6 13.8
7 14 14.2 14.4 14.6 14.8 15 15.2 15.4 15.6 15.8
8 16 16.2 16.4 16.6 16.8 17 17.2 17.4 17.6 17.8
9 18 18.2 18.4 18.6 18.8 19 19.2 19.4 19.6 19.8
10 20 20.2 20.4 20.6 20.8 21 21.2 21.4 21.6 21.8
11 22 22.2 22.4 22.6 22.8 23 23.2 23.4 23.6 23.8
12 24 24.2 24.4 24.6 24.8 25 25.2 25.4 25.6 25.8
13 26 26.2 26.4 26.6 26.8 27 27.2 27.4 27.6 27.8
14 28 28.2 28.4 28.6 28.8 29 29.2 29.4 29.6 29.8
page 19 15 30 30.2 30.4 30.6 30.8 31 31.2 31.4 31.6 31.8
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MENU Table 1 (cont’d): WBC Estimate Chart (40x High)

BACK 1 WBC = 2.0 x 103/µL
.0 .1 .2 .3 .4 .5 .6 .7 .8 .9
NEXT
16 32 32.2 32.4 32.6 32.8 33 33.2 33.4 33.6 33.8
EXIT
17 34 34.2 34.4 34.6 34.8 35 35.2 35.4 35.6 35.8
18 36 36.2 36.4 36.6 36.8 37 37.2 37.4 37.6 37.8
19 38 38.2 38.4 38.6 38.8 39 39.2 39.4 39.6 39.8
20 40 40.2 40.4 40.6 40.8 41 41.2 41.4 41.6 41.8
21 42 42.2 42.4 42.6 42.8 43 43.2 43.4 43.6 43.8
22 44 44.2 44.4 44.6 44.8 45 45.2 45.4 45.6 45.8
23 46 46.2 46.4 46.6 46.8 47 47.2 47.4 47.6 47.8
24 48 48.2 48.4 48.6 48.8 49 49.2 49.4 49.6 49.8
25 50 50.2 50.4 50.6 50.8 51 51.2 51.4 51.6 51.8
26 52 52.2 52.4 52.6 52.8 53 53.2 53.4 53.6 53.8
27 54 54.2 54.4 54.6 54.8 55 55.2 55.4 55.6 55.8
28 56 56.2 56.4 56.6 56.8 57 57.2 57.4 57.6 57.8
29 58 58.2 58.4 58.6 58.8 59 59.2 59.4 59.6 59.8
30 60 60.2 60.4 60.6 60.8 61 61.2 61.4 61.6 61.8
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MENU Reference
BACK Frankel S, Reitman S, Sonnevirth A, eds. Clinical Laboratory Methods and Diagnosis. St. Louis,
MO: C.V. Mosby Company; 1970:505.
NEXT
Limitations
EXIT
1. This procedure is based on using the 40x high dry objective. Using a different objective may alter
results.
2. This estimate is based on using a planar objective. A nonplanar objective may give different
results.
3. Uneven distribution of the WBCs on the smear will adversely affect the estimate. Do not perform
this procedure unless a properly prepared smear is available.
4. The chart in Procedure B will not be applicable to all laboratories. Factors that may affect the
efficacy of the chart include:
• manner of blood smear preparation
• area on the smear for blood cell estimate
• type of objective used
• the blood specimen’s hematocrit
page 21
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MENU References
BACK Frankel S, Reitman S, Sonnenvirth A, eds. Clinical Laboratory Methods and Diagnosis. Saint Louis,
NEXT
Lewis E, Koepke J. Hematology Laboratory Management Practice. Linacre House, Jordan Hill,
EXIT Oxford: Butterworth-Heinemann, Ltd; 1995:16.
O’Connor B. A Color Atlas and Instruction Manual of Peripheral Blood Cell Morphology.
Baltimore, MD: Williams and Wilkins; 1984:20.
Davis; 1992:1547.
page 22
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MENU RBC INTERFERENCES

BACK
NEXT Hemolyzed Specimens
EXIT Definition
Hemolyzed specimens are those in which red cells are being destroyed, thus liberating their hemoglo-
bin. This process may be caused either by in vivo conditions—such as hemolytic disease of the
newborn, other autoimmune diseases, or transfusion reactions. In vitro conditions such as poor
drawing or handling of the blood specimen can also cause hemolysis.
The CELL-DYN 3000/3500/3700 directly measured parameters affected by hemolysis are RBCs and
PLTs. The RBC count may be decreased, and the PLT count may be elevated.
page 23
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BACK • Redraw the blood specimen, if possible. If this is not possible, follow your laboratory protocol.
NEXT • If redrawing is not possible, correct results as follows:

HGB x 2.98 = cHCT
EXIT
12.1 x 2.98 = 36.1
(cHCT ÷ MCV) x 10 = cRBC
36.1 ÷ 86.2 = 4.18
References
Davis; 1992:505, 193–221.
Kalache GR, Sartos MM, Hughes WG. The indirect estimation of hemoglobin concentration in whole
blood. Pathology. 1991;115–117.
page 24
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SUSPECT
MENU
WBC 5.17 K /µL (WOC)
BACK NEU 3.81 73.8 %N
LYM .945 18.3 %L
NEXT MONO .310 5.99 %M NRBC
EOS .088 1.59 %E
EXIT
BASO .020 .388 %B
RBC 4.13 M /µL

HGB 12.2 g/dL
HCT 35.5 %
MCH 29.4 pg
MCHC 34.3 g/dL
RDW 12.4 %
PLT 323. K /µL

MPV 10.2 fL
Sequence 5691: Normal Specimen without Hemolysis
page 25
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Param Set 1 Limit Set 1

MENU
SUSPECT
BACK WBC 5.22 K /µL (WOC)
NEU 3.79 72.7 %N
NEXT LYM 1.02 19.6 %L
MONO .304 5.83 %M NRBC
EXIT
EOS .077 1.48 %E
BASO .021 .408 %B
RBC 3.38 M /µL

HGB 12.1 g/dL
HCT 29.2 %
MCH 35.7 pg
MCHC 41.4 g/dL
RDW 12.9 %
PLT 260. K /µL

MPV 10.7 fL
Sequence 5692: Hemolyzed Specimen
page 26
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MENU Spuriously Decreased Hematocrits

BACK Definition
NEXT Spuriously (falsely) decreased hematocrits occur because an insufficient sample has been drawn into
EXIT the EDTA tube. This may result in shrinkage of the RBCs.
Shrinkage of the RBCs falsely decreases the HCT and falsely increases the MCHC.
Corrective Actions
• Check the sample volume in the EDTA tube. Incomplete filling of a blood collection tube should
be noted.
• If necessary, follow your laboratory protocol and/or redraw the patient, verifying that sufficient
sample volume has been collected.
Reference
Turgeon ML. Clinical Hematology Theory and Procedures. Boston, MA: Little, Brown and
Company; 1993:22, 348.
page 27
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MENU
WBC 6.64 K /µL
BACK NEU 4.20 63.3 %N
LYM 1.84 27.7 %L
NEXT MONO .452 6.81 %M
EOS .094 1.41 %E
EXIT
BASO .055 .835 %B
RBC 4.37 M /µL

HGB 12.7 g/dL
HCT 37.6 %
MCV 86.2 fL
MCH 29.1 pg
MCHC 33.7 g/dL
RDW 12.9 %
PLT 306. K /µL

MPV 8.51 fL
Sequence 5658: Insufficient Sample
page 28
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NORMAL
MENU
WBC 7.19 K /µL
BACK NEU 4.49 63.3 %N
LYM 2.11 27.7 %L
NEXT MONO .471 6.81 %M
EOS .086 1.41 %E
EXIT
BASO .036 .835 %B
RBC 4.75 M /µL

HGB 13.8 g/dL
HCT 41.4 %
MCV 87.2 fL
MCH 29.0 pg
MCHC 33.3 g/dL
RDW 12.8 %
PLT 327 K /µL

MPV 8.32 fL
Sequence 5659: redrawn sample 5658
page 29
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MENU Cold Agglutinins

BACK Definition
NEXT Cold agglutinins are autoantibodies which cause RBCs to agglutinate (clump) when the temperature
EXIT of blood is lowered below 37˚C.
The presence of autoantibodies may be due to a chronic idiopathic disease state. Autoantibodies may
also appear in cases of HIV infection or other immunodeficiency disorders, as well as Mycoplasma
pneumoniae infections. Secondary forms of cold agglutinin syndrome may accompany malignant
diseases of lymphoid origin.
RBC agglutination may result in a falsely decreased RBC count and a greatly increased MCH and
MCHC.
Corrective Actions
• If the MCHC is >36.0 and the plasma is not lipemic and does not indicate hemolysis, warm the
sample to 37˚C for a minimum of 15 minutes (stronger cold agglutinins may need up to
45 minutes incubation).
• Repeat the sample and determine if the MCHC is within the laboratory’s normal range.
page 30
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MENU • Examine a stained smear for the presence of agglutinated RBCs and follow your laboratory
protocol.
BACK
• If warming is not possible or is ineffective, correct results as follows:
NEXT
HGB x 2.98 = cHCT
EXIT 9.09 x 2.98 = 27.1
(cHCT ÷ MCV) x 10 = cRBC
27.1 ÷ 87.9 = 3.08
Reference
Kalache GR, Sartor MM, Hughes WG. The indirect estimation of hemoglobin concentration in whole
Wintrobe M, Lee G, Boggsy D, Bithell L, Athens J, Foerster J. Wintrobe’s Clinical Hematology.
Philadelphia, PA: Lea & Febiger; 1993:1182.
page 31
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SUSPECT
MENU
WBC 18.6 K /µL
BACK NEU 16.0 85.9 %N BAND
LYM .846 4.55 %L
EOS .032 .170 %E
EXIT
BASO .144 .772 %B
RBC 2.26 M /µL

HGB 9.09 g/dL
HCT 19.9 %
MCH 40.3 pg
MCHC 45.8 g/dL
RDW 14.9 %
PLT 51.7 K /µL

MPV 10.3 fL
Sequence 753: Cold Agglutinins
page 32
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SUSPECT
MENU
WBC 19.6 K /µL
BACK NEU 16.9 86.2 %N BAND
EOS .031 .160 %E
EXIT
BASO .350 1.78 %B
RBC 3.08 M /µL

HGB 9.13 g/dL
HCT 26.1 %
MCV 84.5 fL
MCH 29.6 pg
MCHC 35.0 g/dL
RDW 15.4 %
PLT 63.2 K /µL

MPV 11.6 fL
Sequence 761: Warmed Sample 753
page 33
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MENU
BACK
NEXT
EXIT
Cold Agglutinins
(reprinted with permission from O’Connor BH. A Color Atlas and
Instructional Manual of Peripheral Blood Cell Morphology. Baltimore,
MD: Williams and Wilkins; 1984:199.)
page 34
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MENU Lyse-resistant RBCs

BACK Definition
NEXT Lyse-resistant RBCs resist the conversion of hemoglobin to cyanmethemoglobin in the presence of
EXIT potassium cyanide or resist osmotic equilibration with sheath reagent.
Lyse-resistant RBCs may interfere in the hemoglobin and/or in the WBC optical count (WOC).
The presence of abnormal hemoglobins such as Hgb S, Hgb C, and Hgb F, or the presence of
abnormal or increased proteins due to a disease state such as multiple myeloma can render the RBCs
resistant to lysis. In some cases of multiple myeloma, the protein may also cause abnormal light
scatter during the WOC measurement.
page 35
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BACK • If a sample has a WBC flag and a suspect NRBC flag, check the size/complexity scatterplot for
interference at the threshold below the lymphocytes. On the CELL-DYN 3500/3700, verify that
NEXT the WOC value is greater than the WIC value.
EXIT • Rerun the sample in the Resistant RBC mode.
• If the WBC flag disappears, the WBC count is reportable. If the WBC flag recurs, the WBC count
must be verified by following your laboratory protocol or using the WBC Estimation Using
Peripheral Smear procedure (see page 17).
NOTE: Other flags may still be present.
References
Joyner RE, Brooks MJ. Evaluation of the automated leucocyte count and differential from the
Cell-Dyn® 3500 in sickle cell disease. Clinical and Laboratory Haematology. 1995;17:329–333.
Dörner K, Schulze S, Reinhardt M, Seeger H, Van Hove L. Improved automated leucocyte counting
and differential in newborns achieved by the haematology analyser CELL-DYN 3500™. Clinical
and Laboratory Haematology. 1995;17:23–30.
page 36
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Specimen ID Feb 08 1994 12:48

MENU
Patient Operator ID 413
BACK Sex Sequence # 1708
Dr Open Sampler
NEXT Param Set 1 Limit Set 1
SUSPECT
EXIT WBC 14.5 K /µL (WIC)WBC
NEU 2.48 17.1 %N
MONO 1.44 9.97 % M RRBC
EOS .735 5.08 %E
BASO 8.29 57.3 % B DFLT (NLMEB)
RBC 4.02 M /µL

HGB 12.2 g/dL
HCT 34.1 %
MCV 84.8 fL
MCH 30.5 pg
MCHC 35.9 g/dL
RDW 15.4 %
PLT 74.6 K /µL

MPV fL LURI
Sequence 1708: Resistant RBCs

page 37 continued on next page
of 88
MENU
BACK
NEXT INTERPRETATION MANUAL DIFFERENTIAL RBC MORPHOLOGY
---------------------WBC-----------------------------------RBC---------------------------PLT-------------------------
EXIT NEU 45 META NORMAL MICRO +
SUSPECTED ABNORMAL POPULATIONS:
WBC Count Alert Nucleated RBC PLT Lower Region Interference
WBC Diff Alert PLT Upper Region Interference BAND 2 MYELO PLYCHROM MACRO +
Variant Lymphocytes
LYMPH 19 PRO HYPCHROM ANISO
USER-DEFINED ABNORMALITIES:
Monocytosis Thrombocytopenia
MONO 15 BLAST POIK + BASOSTIP
ATL burr +
Eosinophilia
Basophilia
EOSIN 14 VAR LYM 5 TARGET + oval +
BASO TOXGRAN SPHERO NRBC
COMMENT:
DIFF BY DATE
Sequence 1708: Resistant RBCs (cont’d)
page 38
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DIAGNOSTIC MENU
MENU
Ready
BACK
12312.0
NEXT
10773.0 LYSE RESISTANT RBC
EXIT
9234.0
7695.0
6156.0
4617.0
3078.0
1539.0
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5
WOC CNT GRAPH
page 39 Sequence 1708

of 88
Specimen ID
MENU
Res.RBC
BACK Sex
Dr
NEXT Parameter Set 1 Limit Set 1
SUSPECT
EXIT WBC 14.3 K /µL (WIC)
NEU 5.64 39.5 %N BAND
LYM 4.01 28.1 %L
MONO 2.56 17.9 %M RRBC
EOS 1.76 12.3 %E
BASO .301 2.11 %B DFLT (NLMEB)
RBC 4.11 M /µL

HGB 12.2 g/dL
HCT 35.0 %
MCV 85.1 fL
MCH 29.7 pg
MCHC 34.9 g/dL
RDW 15.0 %
PLT 77.7 K /µL

MPV >>>> fL LURI
Sequence 1709: Sequence 1708 in Resistant RBC Mode

page 40 continued on next page
of 88
INTERPRETATION
MENU ---------------------WBC-----------------------------------RBC---------------------------PLT-------------------------
SUSPECTED ABNORMAL POPULATIONS:

BACK WBC Diff Alert Nucleated RBC PLT Data Overrange
Bands PLT Lower Region Interference
WIC WOC Delta Alert PLT Upper Region Interference
NEXT
USER-DEFINED ABNORMALITIES:
EXIT Monocytosis Thrombocytopenia
Eosinophilia
Basophilia
Sequence 1709: Sequence 1708 in Resistant RBC Mode (cont’d)
page 41
of 88
MENU Hyperglycemia
BACK Definition
NEXT Hyperglycemia is an elevation of blood glucose (sugar) and is most commonly encountered in diabetes
EXIT mellitus.
Increased blood glucose levels may falsely elevate the electronically determined MCV and HCT,
apparently from the osmotic swelling and subsequent shrinking of hyperglycemic erythrocytes when
they are mixed with the diluent used to measure hematologic parameters. When the RBCs are sus-
pended in a diluent with a lower glucose concentration, water moves across the cell membrane into
the cells first, and glucose equilibrates later. If the MCV measurement takes place before equilibra-
tion while the cells are still swollen, the MCV and HCT may be falsely elevated.
page 42
of 88

BACK • Dilute a well-mixed aliquot of whole blood with diluent. (For dilution examples, see page 14.)
• Run the well-mixed dilution as a patient. Correct results for the dilution.
NEXT
or
EXIT
• Perform a spun hematocrit and back-calculate the MCV and MCHC.
• Follow your laboratory’s protocol.
• In the case of hyperglycemia where the MCH is unaffected and the MCV is elevated, the MCV
can be corrected by applying the established factor of 2.98 in the following formula:
cMCV = MCH x 2.98 or cHCT = HGB x 2.98

Hematocrit and MCHC can then be calculated by the following
formula:
HCT = MCV x RBC

10
MCHC = HGB x 100

HCT
Reference
page 43 blood. Pathology. 1991;115–117.
of 88
WBC 4.90 K /µL

MENU
NEU 2.96 60.4 %N
BACK LYM 1.48 30.3 %L
MONO 3.53 7.22 %M
NEXT EOS .082 1.68 %E
BASO .021 .431 %B
EXIT
RBC 3.39 M /µL
HGB 9.74 g/dL
HCT 31.1 %
MCV 91.9 fL
MCH 28.8 pg
MCHC 31.3 g/dL
RDW 15.6 %
PLT 251. K /µL

MPV 10.8 fL
Sequence 5701: Hyperglycemia
page 44
of 88
WBC 5.72 K /µL

MENU
NEU 3.56 62.2 %N
BACK LYM 1.70 29.7 %L
MONO .358 6.27 %M
NEXT EOS .072 1.26 %E
BASO .031 .534 %B
EXIT
RBC 3.95 M /µL
HGB 11.2 g/dL
HCT 34.4 %
MCV 87.1 fL
MCH 28.3 pg
MCHC 32.5 g/dL
RDW 14.1 %
PLT 293. K /µL

MPV 10.9 fL
Sequence 5695: Normal Sample without Hyperglycemia
page 45
of 88
MENU HGB INTERFERENCES

BACK
NEXT Lipemia
EXIT Definition
Hyperlipidemia is an elevation of one or more of the blood lipids that may produce an opalescent to
milky or turbid (lactescent) appearance of serum. It may indicate the usual postprandial rise of an
abnormal elevation of triglycerides or cholesterol or both. Hyperlipidemia is a lactescent appearance
of plasma caused by an increased concentration of triglycerides in either VLDL or chylomicrons.
Erroneously high hemoglobin and erythrocyte indices have been reported for patients with severe
hypertriglyceridemia, patients with hyperchylomicronemia, and patients receiving intravenous
administration of fat emulsions. The turbidity of the plasma falsely elevates a spectrophotometric
hemoglobin reading.
page 46
of 88
SUSPECT
MENU
WBC 1.45 K /µL (WIC) WBC
BACK NEU .026 1.78 %N
LYM .242 16.7 %L
NEXT MONO .089 6.17 %M NRBC
EOS 1.08 74.8 %E
EXIT
BASO .008 .525 %B DFLT (NLMEB)
RBC 1.85 M /µL

HGB 5.71 g/dL
HCT 14.3 %
MCH 30.9 pg
MCHC 39.9 g/dL
RDW 18.5 %
PLT 114. K /µL

MPV fL LURI
Sequence 2762: Hyperlipidemia
page 47
of 88
MENU Corrective Action

BACK Follow your laboratory’s protocol and correct the HGB indices using the following:
NEXT • Plasma Replacement Procedure

• Plasma Hemoglobin Procedure
EXIT • Indirect Estimation Procedure
page 48
of 88
MENU Plasma Replacement Procedure

BACK Purpose
NEXT To correct the hemoglobin value obtained in the presence of lipemia using plasma replacement.
EXIT
Specimen
Fresh whole blood anticoagulated with EDTA.
Procedure
1. Run the well-mixed sample as usual and record the hemoglobin and hematocrit values.
2. Centrifuge an aliquot of whole blood (about 3 mL).
3. After centrifuging, mark on the tube the location where the meniscus of the plasma is.
4. Remove and discard the lipemic plasma.
5. Replace the plasma with equal volume of saline.
6. Mix and run the specimen as before, and record the hemoglobin and hematocrit values.
page 49
of 88
MENU Notes
BACK • The hematocrit values before and after washing should be within ± 5%, which indicates that the
cell concentration was not affected.
NEXT
• Hemoglobin interference is usually recognized when the rule of three is broken and/or an MCHC
EXIT value outside the laboratory’s normal range is observed.
Reference
Gagne C, Auger PL, Moorjani S, Brun D, and Lupien PJ. Effect of hyperchylo-micronemia on the
measurement of hemoglobin. American Journal of Clinical Pathology. 1977;68:584–586.
page 50
of 88
MENU Plasma Hemoglobin Procedure

BACK Purpose
NEXT To correct the hemoglobin value obtained in the presence of lipemia using plasma hemoglobin.
EXIT
Specimen
Procedure
1. Run the well-mixed sample as usual and record the hemoglobin and hematocrit values.
2. Centrifuge an aliquot of the sample at 3000 rpm for 10 minutes.
3. Remove as much of the plasma as possible without disturbing the cells and place into a separate
tube.
4. Run the plasma specimen as a patient specimen and record the hemoglobin value.
NOTE: The instrument will give a Sampling Error or Incomplete Aspiration message.
page 51
of 88
MENU Calculations
BACK Corrected HGB = HGBwhole blood - [(1- HCT*) x HGBplasma]
NEXT * HCT expressed in decimal format.
EXIT
Notes
Hemoglobin interference is usually recognized when the rule of three is broken and/or and MCHC
value outside the laboratory’s normal range is observed. Recalculate the MCH and MCHC using the
corrected HGB value.
References
CELL-DYN 3000 Training Guide.
Jerome SN, Roark MF, Wanser C. Anemia Masked by Triglyceridemia. Denver, CO: Department of
Pathology, General Rose Memorial Hospital and University of Colorado Medical Center; 1974.
page 52
of 88
MENU Indirect Estimation Procedure

BACK Purpose
NEXT To correct the hemoglobin value obtained in the presence of lipemia using indirect estimation. This
EXIT method, which does not require initial RBC lysis, corrects for the falsely elevated HGB due to lipemic
plasma and determines whether an elevated WBC count is altering the HGB value. It is based on the
observation of a constant ratio (2.98) between MCV and MCH (MCV/MCH = 2.98).
Specimen
Procedure
1. Process the blood specimen as usual.
2. Correct the hemoglobin value using the following formula:
Corrected HGB g/dL = MCV x RBC

2.98 x 10
page 53
of 88
MENU Example:
BACK Original values: RBC: 5.00 cHGB = 90 x 5.00

HGB: 17.8 2.98 x 10
NEXT
HCT: 45.0
EXIT MCV: 90 = 15.1 g/dL
3. Recalculate MCH and MCHC using corrected HGB.
Reference
page 54
of 88
MENU PLATELET INTERFERENCES

BACK
NEXT Pseudothrombocytopenia—
EXIT Spurious Platelet Clumping
Definition
Platelet clumping may be due to a cold-reactive platelet agglutinin or to a temperature-independent
agglutinin which is demonstrated in the presence of EDTA. Such agglutinins may also be drug
induced in some cases.
Spuriously low and widely fluctuating platelet counts caused by EDTA-dependent agglutination
and/or satellitism may be observed.
page 55
of 88

BACK • Follow your laboratory protocol.
NEXT • Perform Platelet Estimate procedure.

• Redraw the specimen and run immediately.
EXIT
• Perform a manual platelet count using the laboratory’s Manual Platelet Count procedure.
• Redraw specimen in Sodium Citrate tube and perform Sodium Citrate procedure (see page 77).
• Examine a stained smear for platelet clumps.
NOTE: Estimates and manual counts are not helpful if platelets have already agglutinated.
References
Shreiner DP, Bell WB. Pseudothrombocytopenia: manifestation of a new type of platelet agglutinin.
Blood. 1973;42:542–549.
Veenhoven WA, Van Der Schans CS, Huiges W, Metting-Scherphuis H, Halie MR, Nieweg HO.
Pseudothrombocytopenia due to agglutinins. American Journal of Clinical Pathology.
1979;72:1005–1008.
page 56
of 88
SUSPECT
MENU
WBC 5.36 K /µL (WIC) WBC
BACK NEU 3.45 64.4 %N
LYM 1.43 26.6 %L
NEXT MONO .346 6.45 %M NRBC/RRBC
EOS .077 1.44 %E
EXIT
BASO .060 1.13 %B DFLT (L)
RBC 5.33 M /µL

HGB 15.8 g/dL
HCT 47.2 %
MCV 88.6 fL
MCH 29.7 pg
MCHC 33.5 g/dL
RDW 13.0 %
PLT 24.0 K /µL

MPV 10.7 fL URI
Sequence 189: Clumped Platelets
page 57
of 88
MENU
BACK
NEXT
EXIT
Clumped Platelets
page 58
of 88
MENU Platelet Satellitism

BACK Definition
NEXT Platelet satellitism is usually characterized by in vitro adherence of platelets to neutrophils or mono-
EXIT cytes in blood anticoagulated with EDTA, absence of the phenomenon on fingerstick smears, and
spurious thrombocytopenia. An IgG plasma factor apparently interacts with EDTA, causing the
platelets to adhere to the surface of neutrophils. The mechanism of platelet satellitism may vary from
case to case.
Spuriously low and widely fluctuating platelet counts caused by EDTA-dependent agglutination
and/or satellitism may be observed.
Corrective Actions
• Redraw specimen in sodium citrate using the Sodium Citrate procedure (see page 77).
page 59
of 88
MENU References
BACK Dale NL, Schumacher HR. Platelet satellitism—new spurious results with automated instruments.
Laboratory Medicine. 1982;13:300–304.
NEXT
Shreiner DP, Bell WR. Pseudothrombocytopenia: manifestation of a new type of platelet agglutinin.
EXIT Blood. 1973;42:541–549.
page 60
of 88
WBC 10.0 K /µL

MENU
NEU 3.63 36.2 %N
BACK LYM 4.27 42.6 %L
MONO .750 7.48 %M
NEXT EOS 1.19 11.9 %E
BASO .182 1.81 %B
EXIT
RBC 5.23 M /µL
HGB 14.2 g/dL
HCT 42.7 %
MCV 81.7 fL
MCH 27.2 pg
MCHC 33.3 g/dL
RDW 13.3 %
PLT 24.6 K /µL

MPV 9.80 fL URI
PCT .024 %
PDW 17.8 10 (GSD) PLT on Na Citrate = 34 x 1.1 = 37
Sequence 8849: Platelet Satellitism
page 61
of 88
MENU
BACK
NEXT
EXIT
Platelet Satellitism
page 62
of 88
MENU Giant Platelets

BACK Definition
NEXT Giant platelets exceed 20 fL in size. CELL-DYN 3000/3500/3700 include platelets up to 35 fL.
EXIT Myeloproliferative disorders, such as myelofibrosis, may cause the release of giant platelets.
The platelet histogram may not descend to the baseline near the upper threshold, a suspect URI flag
may be displayed, and/or the MPV may be high or overrange.
Corrective Actions
• Examine a stained smear using the Platelet Estimation procedure.
• Perform a manual platelet count using the laboratory’s Manual Platelet Count procedure.
Reference
Davis; 1992:358, 443.
page 63
of 88
WBC 4.29 K /µL

MENU
NEU 2.11 49.1 %N
BACK LYM 1.50 34.9 %L
MONO .441 10.3 %M
NEXT EOS .214 4.97 %E
BASO .035 .812 %B
EXIT
RBC 4.67 M /µL
HGB 11.9 g/dL
HCT 35.7 %
MCH 25.4 pg
MCHC 33.3 g/dL
RDW 15.9 %
PLT 259. K /µL

MPV 14.2 fL URI
Sequences 4068: Giant Platelets

NOTE: Overlap is equal and opposite. An erroneous count may occur when there is no valley and
the upper threshold cuts an ascending or descending line.
page 64
of 88
MENU
BACK
NEXT
EXIT
Giant Platelets
page 65
of 88
MENU Megakaryocytes
BACK Definition
NEXT Megakaryocytes are precursors of mature platelets. Intact megakaryocytes and megakaryocytic frag-
EXIT ments are occasionally found in the peripheral blood from patients with myeloproliferative diseases
such as chronic myelocytic and megakaryocytic leukemia, and in leukemic myelofibrosis. They are
only rarely observed in peripheral blood smears of normal individuals.
The rarity of the presence of megakaryocytes precludes clinical studies defining their position on the
scatterplot. However, one human case with circulating micro-megakaryocytes has shown that small
megakaryocytes seem to fall within the noise region (N2 Region) and are not counted as WBCs. This
seems logical, since micro-megakaryocytes can be about the size of lymphocytes but have much more
internal complexity than lymphocytes. They fall on the size scale at the lymphocyte level but on the
complexity scale at the neutrophil level. Therefore, mature megakaryocytes would be off the size
scale. Megakaryocyte fragments could fall anywhere along the size scale at the neutrophil complexity
level, but would be expected to be too infrequent to have a clinically significant effect on results.
If megakaryocyte fragments were present in significant numbers and happened to be the same size as
neutrophils, an IG suspect flag could be generated.
page 66
of 88

BACK • Include scattergram interpretation for abnormal cluster in the review
criteria and follow your laboratory’s protocol.
NEXT
• Review a manual smear if abnormal scatterplots are observed.
EXIT
References
Diggs LW, Sturm D, and Bell A. The Morphology of Human Blood Cells. Abbott Park, IL: Abbott
Laboratories; 1985:33.
Janik J. Customer letter dated 6/6/95 from Abbott Diagnostics, Santa Clara, CA.
page 67
of 88
SUSPECT
MENU
WBC 6.66 K /µL
BACK NEU 1.38 20.7 %N BANDS
LYM 4.42 66.5 %L BLAST
NEXT MONO .549 8.25 %M NWBC
EOS 0.00 0.00 %E
EXIT
BASO .300 4.52 %B
RBC 2.30 M /µL

HGB 6.71 g/dL
HCT 20.3 %
MCV 88.0 fL
MCH 29.1 pg
MCHC 33.1 g/dL
RDW 15.6 %
PLT 440. K /µL

MPV 11.7 fL
Sequence 6117: Megakaryocytes

continued on next page
page 68
of 88
MANUAL DIFFERENTIAL RBC MORPHOLOGY

MENU
NEU 4 META 1 NORMAL MICRO +
BACK
BAND 10 MYELO 3 PLYCHROM MACRO +
NEXT LYMPH 60 PRO HYPCHROM ANISO
EXIT e al <— MONO 14 BLAST POIK + BASOSTIP

som orm
n 0 5 +
ab EOSIN VAR LYM TARGET
BASO 0 TOXGRAN SPHERO NRBC 17

COMMENT: Megakaryocyte Frag-8
DIFF BY DATE
Sequence 6117: Megakaryocytes (cont’d)
NOTE: Results in manual differential grid reflect actual manual differential performed from a stained
smear.
page 69
of 88
MENU
BACK
NEXT
EXIT
Megakaryocytes
(reprinted with permission from O’Connor BH. A Color Atlas and
Instructional Manual of Peripheral Blood Cell Morphology. Baltimore,
MD: Williams and Wilkins; 1984:108.)
page 70
of 88
MENU Falsely Elevated/Overrange Platelet Count

BACK Definition
NEXT This section discusses a platelet count that exceeds the linearity limits of the instrument. It may be
EXIT due to interference from a variety of causes, including Howell-Jolly bodies, nucleated RBCs, cellular
debris, microcytic RBCs, bacterial contamination of reagents, schistocytes, or electrical
interference.
Chevrons (>>>>) appear in place of the PLT count (greater than 2 M/µL for CELL-DYN 3500/3700;
greater than 999 K/µL for CELL-DYN 3000).
page 71
of 88

BACK • Follow your laboratory’s protocol and examine a stained smear using the Platelet Estimation
procedure.
NEXT
• If the platelet count appears elevated, pre-dilute the sample with diluent as follows:
EXIT
1. Dilute a well-mixed aliquot of whole blood with diluent.
Examples:
0.5 mL whole blood + 0.5 mL diluent (1:2 dilution) → multiply PLT by 2
2. Run the well-mixed dilution as a patient.
NOTE: The specimen may be run in the Auxiliary Mode on the

CELL-DYN 3500/3700.
page 72
of 88
MENU 3. For specimens run on the CELL-DYN 3000, the PLT result must be multiplied by the dilution
factor before reporting. The Auxiliary Mode on the CELL-DYN 3500/3700 automatically corrects
BACK the PLT count for the dilution factor.
NEXT • If interference occurs due to RBC abnormalities, follow your labor-atory’s protocol and
perform a manual platelet count using the Manual Platelet Count procedure.
EXIT
• If electrical interference is suspected, a suspect LRI may be generated. Troubleshoot the instru-
ment using the troubleshooting section of the appropriate operator’s manual and/or perform
platelet count by an alternative method.
References
Davis; 1992:454–455.
page 73
of 88
SUSPECT
MENU
WBC 78.3 K /µL
BACK NEU 58.0 74.1 %N IG
NEXT MONO 1.68 2.15 %M NWBC
EOS 2.88 3.68 %E
EXIT
BASO .520 .665 %B DFLT (L)
RBC 3.95 M /µL

HGB 8.40 g/dL
HCT 24.8 %
MCH 21.2 pg
MCHC 33.9 g/dL
RDW 38.2 %
PLT >>>> K /µL

MPV 7.68 fL
Comment: PLT count = 3130 K giant plts
Sequence 9568: Overrange Platelet Count
page 74
of 88
WBC 6.66 K /µL

MENU
NEU 4.28 64.3 %N
BACK LYM 1.90 28.6 %L
MONO .328 4.92 %M
NEXT EOS .085 1.27 %E
BASO .063 .952 %B
EXIT
RBC 4.74 M /µL
HGB 14.0 g/dL
HCT 42.1 %
MCV 88.8 fL
MCH 29.5 pg
MCHC 33.2 g/dL
RDW 14.9 %
PLT 241. K /µL

MPV fL
Sequence 7062: Electrical Interference

continued on next page
page 75
of 88
INTERPRETATION MANUAL DIFFERENTIAL RBC MORPHOLOGY

MENU ---------------------WBC-----------------------------------RBC---------------------------PLT-------------------------
SUSPECTED ABNORMAL POPULATIONS: NEU 59 META NORMAL MICRO +

BACK PLT Lower Region Interference
BAND 2 MYELO PLYCHROM MACRO +
NEXT LYMPH 29 PRO HYPCHROM ANISO
EXIT MONO 7 BLAST POIK + BASOSTIP
EOSIN 1 VAR LYM TARGET oval +

BASO 2 TOXGRAN SPHERO NRBC
COMMENT:
DIFF BY DATE
Sequence 7062: Electrical Interference (cont’d)
page 76
of 88
MENU Alternative Platelet Procedures:

BACK
Sodium Citrate Procedure for Platelet Clumping
NEXT Purpose
EXIT Spuriously low and widely fluctuating platelet counts caused by EDTA-dependent agglutination
and/or satellitism may be overcome with the use of a different anticoagulant, such as redrawing in
sodium citrate.
Specimens
Fresh whole blood anticoagulated with sodium citrate (light-blue stopper tube, one part sodium citrate
to nine parts blood).
A well-prepared, stained peripheral blood smear.
page 77
of 88
MENU Procedure
BACK 1. If platelet counts are observed that are unexpectedly low or fluctuating (that is platelet counts fall
as time goes on), platelet numbers should be estimated by examining the blood smear made with
NEXT
EDTA-anticoagulated blood. If platelet clumping or satellitism is observed, a blood smear using
EXIT unanticoagulated blood from a finger stick should be reviewed.
2. If a finger stick smear reveals no platelet agglutination or satellitism, this indicates either a bad
draw or EDTA-dependent spurious agglutination. Redraw a specimen using sodium citrate as the
anticoagulant (light blue stopper tube).
3. Rerun the specimen. Only the platelet results may be used. Multiply the platelet results by 1.1
(corrected for the sodium citrate anticoagulant dilution) and record the new results.
Example:
Sodium citrate platelet count = 158,000

158,000 x 1.1 = 173,800 → 174,000 (corrected count)
page 78
of 88
MENU References
BACK Dale NL, Schumacher HR. Platelet satellitism—new spurious results with automated instruments.
Laboratory Medicine. 1982;13:300–304.
NEXT
Shreiner DP, Bell WR. Pseudothrombocytopenia: manifestation of a new type of platelet agglutinin.
EXIT Blood. 1973;42:541–549.
Veenhouen WA, Van Der Schans GS. Pseudothrombocytopenia due to agglutinins. Blood.
1973,42:541–549.
page 79
of 88

BACK
Platelet Estimation Using Peripheral Smear
NEXT Purpose
EXIT To estimate platelet counts by microscopically examining a stained smear.
Specimen
Wedge smear made from fresh EDTA-anticoagulated blood producing a feathered edge. Stain the
smear according to laboratory protocol.
Procedure A
1. Scan the smear under a 10x (low) objective to ensure even distribution of cells.
2. Examine the smear using a 100x oil objective. Select an area of the smear where the RBCs are
just touching each other, and count the number of platelets in 10 fields.
3. Multiply results by 2000.
page 80
of 88
MENU Procedure B
BACK 1. Follow steps 1 and 2 in Procedure A above.
NEXT 2. Obtain the average platelet count per 100x oil field. Do not truncate the number.
EXIT 3. Obtain the estimated platelet count as follows:
a. Using Table 2 on the next pages, find the whole number in the left
column and the decimal value in the top row.
b. The number at the intersection of these two points is the estimated platelet count.
Example:
If the average number of PLTs/100x field = 5.2,

then the platelet estimate = 104.0 x 103.
page 81
of 88
MENU Table 2: Platelet Estimate Chart (100x Oil)

BACK 1 PLT/Oil Field = 20 x 103/µL (continued)
0.0 .0 .1 .2 .3 .4 .5 .6 .7 .8 .9
NEXT
0 0.0 2 4 6 8 10 12 14 16 18
EXIT
1 20 22 24 26 28 30 32 34 36 38
2 40 42 44 46 48 50 52 54 56 58
3 60 62 64 66 68 70 72 74 76 78
4 80 82 84 86 88 90 92 94 96 98
5 100 102 104 106 108 110 112 114 116 118
6 120 122 124 126 128 130 132 134 136 138
7 140 142 144 146 148 150 152 154 156 158
8 160 162 164 166 168 170 172 174 176 178
9 180 182 184 186 188 190 192 194 196 198
10 200 202 204 206 208 210 212 214 216 218
11 220 222 224 226 228 230 232 234 236 238
12 240 242 244 246 248 250 252 254 256 258
13 260 262 264 266 268 270 272 274 276 278
14 280 282 284 286 288 290 292 294 296 298
page 82 15 300 302 304 306 308 310 312 314 316 318
of 88
MENU Table 2 (cont’d): Platelet Estimate Chart (100x Oil)

BACK 1 PLT/Oil Field = 20 x 103/µL
0.0 .0 .1 .2 .3 .4 .5 .6 .7 .8 .9
NEXT
16 320 322 324 326 328 330 332 334 336 338
EXIT
17 340 342 344 346 348 350 352 354 356 358
18 360 362 364 366 368 370 372 374 376 378
19 380 382 384 386 388 390 392 394 396 398
20 400 402 404 406 408 410 412 414 416 418
21 420 422 424 426 428 430 432 434 436 438
22 440 442 444 446 448 450 452 454 456 458
23 460 462 464 466 468 470 472 474 476 478
24 480 482 484 486 488 490 492 494 496 498
25 500 502 504 506 508 510 512 514 516 518
26 520 522 524 526 528 530 532 534 536 538
27 540 542 544 546 548 550 552 554 556 558
28 560 562 564 566 568 570 572 574 576 578
29 580 582 584 586 588 590 592 594 596 598
30 600 602 604 606 608 610 612 614 616 618
page 83
of 88
MENU Reference
BACK Frankel S, Reitman S, Sonnevirth A, eds. Clinical Laboratory Methods and Diagnosis. St. Louis,
NEXT
Calculation
EXIT
Platelet estimate = number of platelets seen in 10 fields x 2000
Note
If the RBC count is much lower than normal, the platelet estimate needs to be adjusted accordingly.
That is, if the RBC count is half of normal, divide the platelet estimate by 2. If the RBC count is 1/3
of normal, divide the platelet estimate by 3.
Limitation
This procedure is for estimating platelet counts only. Do not use for absolute quantification. Table 2
will not be applicable to all laboratories. Factors that may affect the efficacy of the chart include
manner of blood smear preparation, area on the smear for blood cell estimate, type of objective used,
and the blood specimen’s hematocrit.
page 84
of 88
MENU References
BACK Frankel S, Reitman S, Sonnenwirth A. Clinical Laboratory Methods and Diagnosis. St. Louis, MO:
C.V. Mosby Company;1970:505.
NEXT
EXIT Davis; 1992:547.
Lewis E, Koepke J. Hematology Laboratory Management and Practice. Jordon Hill, Oxford:
Butterworth-Heinemann, Ltd. Linacre House; 1995: 16.
O’Connor B. A Color Atlas and Instruction Manual of Peripheral Blood Cell Morphology.
Baltimore, MD: Williams and Wilkins; 1984.
page 85
of 88

BACK
Manual Platelet Count
NEXT Principle
EXIT This method is used to count platelets when there is interference with automated platelet counts. It is
based on the dilution of whole blood in a solution of 1% ammonium oxalate. The RBCs lyse, leaving
leukocytes, reticulocytes, and platelets. The platelets appear as dark bodies which are easily distin-
guished from “ghost” cells (RBCs), leukocytes, and reticulocytes.
Specimen
Fresh whole blood collected in EDTA or directly into the B-D Unopette™ capillary pipette.
Procedure
1. Clean and dry an Improved Neubauer hemocytometer and hemocytometer cover glass.
2. Mix the EDTA sample by inversion 20 times or on a mechanical mixer for 2 to 3 minutes.
3. Puncture the diaphragm of two Unopette™ WBC/platelet count reservoirs using the pointed plastic
shield of the capillary pipette.
page 86
of 88
MENU 4. Fill the capillary tube with well-mixed whole blood collected in EDTA (or directly from a finger
stick puncture wound) and transfer to the reservoir.
BACK
5. Allow the dilution to incubate according to the manufacturer’s instructions for RBC lysis to occur.
NEXT
6. Fill, by capillary action, two sides of a carefully cleaned hemocytometer, a separate dilution on
EXIT each side.
7. Place the hemocytometer on moistened towel or gauze in a petri dish, and allow it to stand
30 minutes to permit cells to settle.
8. Using 400x magnification, count all the platelets in all 25 small squares within the large center
square using a phase microscope.
9. Calculate the final platelet count for each side of the chamber (the replicate counts should agree
with ± 5%):
(# platelets counted on one side) x 100 x 10 x 1 = total platelet count

(dilution (depth) (area)
factor)
page 87
of 88
MENU References
BACK Akwaris R. Spuriously elevated platelet counts due to microspherocytosis. American Journal of
Clinical Pathology. 1981;77:220–221.
EXIT
Brecker G, Cronkite EP. Morphology and enumeration of human blood platelets. Journal of Applied
Physiology. 1950;3:365.
Davis; 1992.
Product circular for Becton-Dickinson Unopette™ Microcollection System, WBC/Platelet
Determination for Manual Methods.
page 88
of 88
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INTERFERING SUBSTANCES
IN HEMATOLOGY
RESOURCE BOOK
CELL-DYN® 3000/3500/3700 Series Systems

Abbott Laboratories
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Copyright 1998, 2001 Abbott Laboratories
All rights reserved. No part of this program may be reproduced in any form
without written permission from Abbott Laboratories.
The material contained herein, including text, charts, graphs, figures, tables,
etc., is not to be used for reference, but is meant for training purposes only.
It is to be distributed exclusively by representatives of Abbott Diagnostics
Division, Abbott Laboratories.
CELL-DYN is a registered trademark of Abbott Laboratories.
All pictures (except where noted) are property (and were provided by the
Hematology Technical Support Specialist Group) of Abbott Laboratories.
Interfering Substances in Hematology Resource Book CELL-DYN® Systems

69927-101 November 2001

Interfering Sub in Hematology

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MENU Using the Navigation Buttons

MENU WBC INTERFERENCES

RBC 4.09 M /µL

Sequence 2117: Fragile WBCs

WOC CNT GRAPH

MENU Clumped WBCs

MENU Corrective Actions

RBC 3.63 M /µL

PLT 201. K /µL

Sequence 6858: Clumped WBCs

MENU WBC Count Overrange

MENU Corrective Actions

RBC 3.21 M /µL

PLT 64.8 K /µL

Sequence 1394: WBC Count Overrange

RBC .355 M /µL

PLT 4.21 K /µL

Sequence 1397: 1:10 Dilution of Sequence 1394

MENU WBC Estimation Using Peripheral Smear

MENU Table 1: WBC Estimate Chart (40x High)

MENU Table 1 (cont’d): WBC Estimate Chart (40x High)

MENU RBC INTERFERENCES

MENU Corrective Actions

NEXT • If redrawing is not possible, correct results as follows:

RBC 4.13 M /µL

PLT 323. K /µL

Sequence 5691: Normal Specimen without Hemolysis

Param Set 1 Limit Set 1

RBC 3.38 M /µL

PLT 260. K /µL

Sequence 5692: Hemolyzed Specimen

MENU Spuriously Decreased Hematocrits

RBC 4.37 M /µL

PLT 306. K /µL

Sequence 5658: Insufficient Sample

RBC 4.75 M /µL

PLT 327 K /µL

Sequence 5659: redrawn sample 5658

MENU Cold Agglutinins

RBC 2.26 M /µL

PLT 51.7 K /µL

Sequence 753: Cold Agglutinins

RBC 3.08 M /µL

PLT 63.2 K /µL

Sequence 761: Warmed Sample 753

MENU Lyse-resistant RBCs

MENU Corrective Actions

Specimen ID Feb 08 1994 12:48

RBC 4.02 M /µL

PLT 74.6 K /µL

Sequence 1708: Resistant RBCs

Sequence 1708: Resistant RBCs (cont’d)

WOC CNT GRAPH

page 39 Sequence 1708

RBC 4.11 M /µL

PLT 77.7 K /µL

Sequence 1709: Sequence 1708 in Resistant RBC Mode

SUSPECTED ABNORMAL POPULATIONS:

Sequence 1709: Sequence 1708 in Resistant RBC Mode (cont’d)

MENU Corrective Actions

cMCV = MCH x 2.98 or cHCT = HGB x 2.98

HCT = MCV x RBC

MCHC = HGB x 100