XN-1000

AUTOMATED HEMATOLOGY ANALYZER

PURPOSE OF THE EXAMINATION

The Sysmex® XN-1000 is automated hematology analyzers for in vitro diagnostic use in
screening patient populations found in clinical laboratories.
This instrument does Complete blood count(CBC) & enables quantitative, identification,
and existence ratio analysis and flagging of tangible components of blood and body fluid
(red blood cells, white blood cells, platelets and other cells) by means of electrical
impedance, laser light scattering, and dye bonding.
Complete blood count is the study of all blood cells in circulation and the estimation of
Hemoglobin in the red blood cells. It is a useful test in the case of Anemia, renal
insufficiency, Leukemia, viral infection and bleeding disorder (Coagulopathies) etc. It is
a very useful test not only of diagnostic significance but also for prognosis and
monitoring the therapy.

PRINCIPLE AND METHOD OF THE PROCEDURE USED FOR

EXAMINATION

The XN1000 performs analysis of WBCs and reticulocytes with an optical detector block
based on the flow cytometry method, using a semiconductor laser.

RBCs and platelet count are analyzed by the RBC detector using the Hydro Dynamic
Focusing method. The hematocrit (HCT) is calculated via the RBC pulse height detection
method.

Hemoglobin (HGB) is analyzed by the HGB detector based on the SLS hemoglobin
detection method.

Analysis data is displayed on the IPU.

Functional Description

1. Detection Principle

This instrument performs hematology analyses according to the Hydro Dynamic

Focusing (DC Detection), flow cytometry method (using a semiconductor laser), and
SLS-hemoglobin method.

2. Hydro Dynamic Focusing (DC Detection)

Inside the detector, the sample nozzle is positioned in line with the center of the
aperture. After diluted sample is forced from the sample nozzle into the conical chamber,
it is surrounded by front sheath reagent and passes through the aperture center.
Passage of cells through the centre of the aperture ensures optimal shape of the cell
signals.
After passing through the aperture, the diluted sample is sent to the catcher tube. This
prevents the blood cells in this area from drifting back into the sensing zone which could
generate false platelet pulses.

The Hydro Dynamic Focusing method improves blood count accuracy and
reproducibility.

3. Flow Cytometry Method Using Semiconductor Laser

Cytometry is used to analyze physiological and chemical characteristics of cells and

other biological particles. Flow cytometry is used to analyze those cells and particles as
they flow through an extremely small pathway.

A blood sample is aspirated, measured, diluted to the specified ratio, and stained. The
sample is then fed into the flow cell.

This Sheath mechanism improves cell count accuracy and reproducibility. Since the
blood cell particles pass in a line through the centre of the flow cell, the generation of
abnormal blood pulses is prevented and flow cell contamination is reduced.

A semiconductor laser beam (wavelength: 633 nm) is focused onto the blood cells
passing through the flow cell. The forward scattered light & lateral scattered light are
received by photodiodes, and the side fluorescent light received by the avalanche
photodiode tubes. This light is converted into electrical pulses, thus making it possible to
obtain blood cell information.

A. Forward Scattered Light and Lateral Scattered Light

When an obstacle such as particle passes through the light beam, the light scatters in
various directions. By detecting the scattered light, it is possible to obtain information on
cell size and material properties.

Likewise, when a laser beam is focussed onto blood cells, light scattering occurs. The
intensity of the scattered light depends on factors such as particle diameter and viewing
angle. This The XN1000 detects forward scattered light, which provides information on
blood cell size; and side scattered light, which provides information on the cell interior
(such as the size of the nucleus).

B. Lateral Fluorescent Light

When light is focused onto fluorescent material, such as stained blood cells, light of
longer wavelength than the original light is produced. The intensity of the fluorescent
light increases as the concentration of the stain becomes higher. By measuring the
intensity of the fluorescence emitted, you can obtain information on the degree of blood
cell staining. Fluorescent light is emitted in all directions; the XN 1000 detects the
fluorescent light that is emitted sideways.
SLS – Hemoglobin Method
In the past, the mainstream methods for automatically measuring hemoglobin have been
cyanmethemoglobin method and oxyhemoglobin method. These methods both have
advantages and disadvantages when they are used with a large, fully automatic
instrument such as the XN 1000.

The cyanmethemoglobin method was recommended by the International Council for

Standardization in Haematology (ICSH) in 1966 as an international standard method.
But since its hemoglobin conversion speed is slow and multiple-sample processing is an
assumed requirement, this method is not really appropriate for automatic measuring.
Moreover, since it uses cyanide compounds, which are making reagents poisonous, the
liquid waste must be treated, making the method undesirable from an environment
perspective. Currently, this is not an appropriate analysis method, particularly for a large
fully automatic instrument that discharges large amounts of liquid waste.

In contrast, the hemoglobin conversion speed of the oxyhemoglobin method is fast, as

blood hemoglobin is instantly converted into oxyhemoglobin. Since it does not use
poisonous substances such as cyanide, it is suitable method for performing automatic
analyses. It cannot, however, convert methemoglobin into oxyhemoglobin, which is not a
problem for normal human blood, but will result in inaccurately low values for samples
that contain large amounts of methemoglobin, such as control blood samples.

The SLS-hemoglobin method is an analysis method that makes use of the advantages
of the two aforementioned methods.

As with the oxyhemoglobin method, the hemoglobin conversion speed of the SLS-
hemoglobin method is fast and the method does not use poisonous substances, making
it a suitable method for automation.
Since it can be used to measure methemoglobin, it can also accurately measure blood
containing methemoglobin, such as control blood.

In the SLS-Hemoglobin method, surfactants lyse the red blood cell membrane releasing
hemoglobin. The globin group of the hemoglobin molecule is altered by the hydrophilic
alkyl group of sodium Lauryl Sulfate. This induces the conversion of hemoglobin from
the ferrous (Fe2+) to ferric (Fe3+) state forming met-hemoglobin, which combines with
Sodium Lauryl Sulfate to become SLS-Hb hemicrome molecule.

On XT-1800i RET and PLT-O are not detected. However, the same operation principle is
used as in XT-2000i.

WBC analysis

WNR channel

The WNR channel is primarily a channel to count the white blood cells and nucleated red
blood cells.
By flow cytometry method using a semiconductor laser, a two-dimensional scattergram
is plotted, with the X-axis representing the intensity of the side fluorescent light (SFL),
and the Y-axis representing the intensity of the forward scattered light (FSC).
This scatter gram displays groups of nucleated red blood cells, basophil,
non-basophil white blood cells, and debris (hemolysed red blood cells and platelets).

WDF channel
SFL
The WDF channel is primarily a channel for classifying
white blood cells. By flow cytometry method using a
semiconductor laser, a Lymphocytes Monocytes
two-dimensional scattergram is plotted, with the X-axis
representing the intensity of the side scattered light Basophils + Neutrophils
(SSC) and the Y-axis representing the intensity of the Eosinophils
side fluorescent light (SFL).
This scatter gram displays groups of lymphocytes,
monocytes, eosinophils, basophils + neutrophils, and SSC
debris.

RBC / PLT particle size distribution Analysis

RBC particle size distribution

The RBC (red blood count) is calculated as a particle count between two discriminators
(lower discriminator (LD) and upper discriminator (UD)), which are automatically set up in
the ranges of 25 - 75 fL and 200 - 250 fL, respectively.
The particle size distribution is checked for abnormal relative frequencies at each
discriminator level, existence of more than one peaks, and abnormal distribution
width.
In this instrument, the RBC distribution width (RDW) is expressed in the following two
ways.

RDW-SD
With the peak height assumed to be 100%, the distribution width at the 20% frequency
level is RDW-SD.
The unit used is femtoliter (fL) (1 fL = 10-15 L).

100%

20%

RDW

RDW-CV
With points L1 and L2 found at a frequency of 68.26% of 68.26% of total distribution area

(L1) (L2)
the total distribution area, RDW-CV is calculated from the following equation:

L2 - L1
RDW-CV (%) = X 100
L2 + L1

PLT particle size distribution

The PLT (platelet count) is calculated as a particle count between two discriminators
(lower discriminator (LD) and upper discriminator (UD)), which are automatically set up in
the ranges of 2 - 6 fL and 12 - 30 fL, [Link] particle size distributions are
checked for abnormalities, including abnormal relative frequencies at the lower
discriminator, abnormal distribution widths, and the existence of more than one peak.

PDW (Calculated distribution width of platelets)

100% P-LCR
With the peak height assumed to be 100%, the distribution
width at the 20% frequency level is PDW.
The unit used is femtoliter (fL) (1 fL = 10-15 L).

P-LCR (Platelet-Large Cell Ratio) 20%

The P-LCR is the ratio of large platelets from the 12 fL (LD) (12 fL) (UD)
PDW
discriminator or larger. It is calculated as a ratio comparing
the number of particles between the fixed discriminator and
UD, to the number of particles between LD and UD.

MPV (Mean Platelet Volume)

The MPV is calculated from the following equation:

PCT (%)
MPV(fL) = X 10000
PLT (x 103/µL)

PCT is called the platelet hematocrit or platelet volume ratio, and is weighted toward the
PLT frequency.

RET (reticulocyte) Analysis

RET Channel
By flow cytometry method using a semiconductor laser, a two-dimensional scattergram
is plotted, with the X-axis representing the intensity of the side fluorescent light (SFL),
and the Y-axis representing the intensity of the forward scattered light (FSC).
This scattergram displays groups of reticulocytes, mature red blood cells and platelets.
The scattergram is divided into three RET zones based on the intensity of the
fluorescent light, and the ratio of the reticulocytes in each zone to the total number of
reticulocytes is calculated.

Reticulocyte Ratio:

Particle count in reticulocyte zone

RET% = x 100
Particle count in mature RBC zone + Particle count in reticulocyte zone

RET (%) X RBC

RET (#) =
100

Immature Reticulocyte Fraction: IRF = MFR + HFR

RET- He (Reticulocyte Hemoglobin equivalent):

The RET- He is a unique parameter developed by Sysmex that is derived using the
reticulocytes scattered light signals.

 MCV calculation: MCV (Mean Cell Volume) is calculated using the Hct (hematocrit)
and the RBC count. The MCV is displayed and printed in femtoliter (fL).
Hct
MCV (fl) = X 10
RBC

MCH and MCHC Calculations

 MCH calculation: MCH (Mean Cell Hemoglobin) is calculated from the Hgb
(hemoglobin) value and the RBC count and describes the average weight of
hemoglobin in a red cell. The calculation for MCH is:

Hgb
MCH (pg) = X 10
RBC
 MCHC calculation: MCHC (Mean Cell Hemoglobin Concentration) is calculated using
the Hgb and Hct values and describes the average concentration of hemoglobin in
the red blood cells. The calculation for MCHC is:
 Hgb
MCHC (g/dL) = x 100
Hct

Analysis Modes

A. Performing System Analysis for Whole Blood:

The sampler automatically mixes, aspirates, and analyzes samples without
removing their caps. Up to 50 samples can be loaded at a time and analyzed
automatically.

B. Manual analysis:

1. Manual Mode
In manual mode, after mixing the sample manually the caps of sample tube
are manually removed and each sample is manually loaded after removing cap in
tube holder.

2. Pre Diluted Mode

In capillary mode, an analysis is performed after manually diluting the sample
to 1:7 dilutions (40ul of blood with 240ul of cellpack DCL). This mode is used
for analyzing a minute amount of blood collected from the earlobe or fingertip.
The sample is manually loaded after removing cap in tube holder and the
obtained result is automatically multiplied by 7 for reporting, which is thus
comparable to the manual mode.

3. Low WBC mode

The XN-Series provides a measurement mode for increasing measurement

precision for low WBC specimens.
In addition to diluting specimens measured using the WNR and WDF channels
compared to the whole blood mode, LW mode increases the leukocyte
measurement count by lengthening the WDF channel measurement time
(increasing the amount of specimen analyzed). In this way, the domain in which
the analyzer can perform fractionation of low-count leukocytes, is expanded to 10
cells/μL, providing absolute neutrophilic count (ANC) data down to low values, an
important source of information in chemotherapy and stem cell transfusions.
Here analysis volume is 174.6 µL instead of 58.2µL.

Sample Measured Tube Type Sample Aspirated Required Remarks

Aspiration Sample setting sample Sample
Mode volume Volume
Sampler Whole Closed Sampler 88 µL 1 mL
Mode of Blood Rack
Analysis
Open tube Tube 88 µL 300 µL
Holder
Open micro Micro 88 µL 160 µL
tubes Tube
Holder
Manual Pre- Open tube Tube 70 µL 300 µL
Mode of Diluted Holder
Analysis Blood
Open micro Micro 70 µL 140 µL
tubes Tube
Holder
Body Closed Tube 88 µL 1 mL
Fluid Holder
Open tube Tube 88 µL 300 µL
Holder
Open micro Micro 88 µL 160 µL
tubes Tube
Holder

Analysis Parameters

Whole Blood, Low WBC mode, Pre Diluted Mode

The XN-1000 provides results for the following parameters.

WBC Number of all leucocytes

RBC Number of all erythrocytes
HGB Hemoglobin concentration
HCT Hematocrit value: Erythrocyte ratio of total blood volume
MCV Mean erythrocyte volume in total sample
MCH Mean hemoglobin volume per RBC
MCHC Mean hemoglobin concentration of erythrocytes
PLT Number of all platelets
NEUT% Neutrophil Percent
LYMPH% Lymphocyte Percent
MONO% Monocyte Percent
EO% Eosinophil Percent
BASO% Basophil Percent
NEUT# Neutrophil Count
LYMPH# Lymphocyte Count
MONO# Monocyte Count
EO# Eosinophil Count
BASO# Basophil Count
RDW-SD Calculated distribution width of erythrocytes, standard deviation
RDW-CV Calculated distribution width of erythrocytes, coefficient of
variation
PDW Calculated distribution width of platelets
MPV Mean platelet volume
P-LCR Platelet-Large Cell Ratio
PCT Plateletcrit
NRBC # Nucleated RBC Count
NRBC % Nucleated RBC Percentage
IG # Immature Granulocyte Count
IG % Immature Granulocyte Percentage
RET% Reticulocyte Percentage
RET# Reticulocyte Count
IRF Immature Reticulocyte Fraction
LFR Low Fluorescence ratio
MFR Medium Fluorescence ratio
HFR High Fluorescence ratio
RET He Reticulocyte Hemoglobin Equivalent

Body Fluid Mode

WBC-BF Number of all leucocytes

RBC-BF Number of all erythrocytes
MN# Mononuclear Count
PMN# Neutrophil Count
MN% Mononuclear Percent
PMN% Neutrophil Percent
TC-BF# Total Nucleated Cell Count

Analysis Parameters and Channels

Measurement Reagent type Reagent name

channel
Each channel Diluent (concentrated CELLPACK DST
reagent)

Diluent CELLPACK DCL

HGB Hemolyzing agent SULFOLYSER

WNR Hemolyzing agent Lysercell WNR

Stain Fluorocell WNR

WDF Hemolyzing agent Lysercell WDF

 Stain Fluorocell WDF

WPC Hemolyzing agent Lysercell WPC

Stain Fluorocell WPC

RET Diluent CELLPACK DFL

Stain Fluorocell RET

PLT-F Diluent CELLPACK DFL

Stain Fluorocell PLT

PERFORMANCE CHARACTERISTICS

Analysis Range

Parameters Analysis Range Display Range

Whole blood Pre-diluted

0- 0.00 -
White Blood Cell (WBC) 0.0-440x103/µL
100x103/µL 999.99x103/µL
Red Blood Cell (RBC) 0-8.60x106/µL 0-8.60x106/ 0.0 -99.99x106/µL
µL
Hemoglobin Concentration 0.0-26.0
0.0-26.0 g/dL 0.0-30.0 g/dL
(HGB) g/dL
Hematocrit (HCT) 0.0-75.0 % 0.0-75.0 % 0.0-100.0 HCT%

0-1000
Platelet Count (PLT) 0-5000.0x103/µL 0.0-9999x103/µL
x103/µL

RET% 0.00-30.00 % 0.00 -99.99%

0.000-0.7200 0.0000-
RET#
x106/µL 0.9999(106/µL)
0.00 -
NRBC #
999.99x103/µL
 0.0 -9999.9/ 100
NRBC %
WBC
Body Fluid

Parameters Analysis Range Display Range

0.000 -
WBC- BF 0.000-10.000 x103/µL
999.999x103/µL
0.000 -
RBC-BF 0.000 - 5.000 x106/µL
99.999x106/µL
MN% - 0.0 – 100%

PMN% - 0.0 – 100%

TC-BF# 0.000-10.000 x103/µL -

Reproducibility

Reproducibility is based on 20 consecutive replicate runs from one normal, fresh whole-
blood sample without flags.

Reproducibility Specification
Reproducibility / Precision
Parameter Whole Blood
Capillary Mode Condition
Mode
White Blood Cell (WBC) Within 3.0% Within 5.0% 4.0x103/µL or
more
Red Blood Cell (RBC) Within 1.5% Within 4.5% 4.00x106/µL or
more
Hemoglobin Within 1.0% Within 3 % -
Hematocrit (HCT) Within 1.5% Within 4.5% -
Mean Corpuscular
(erythrocyte) Volume Within 1.0% Within 4.5% -
(MCV)
Mean Corpuscular
Within 2.0% Within 4.5% -
Hemoglobin (MCH)
Mean Corpuscular
Hemoglobin Within 2.0% Within 6.0% -
Concentration (MCHC)
Platelet Count (PLT) Within 4.0% Within 12.0% 10x104/µL or more
 Reproducibility / Precision
Parameter Whole Blood
Capillary Mode Condition
Mode
RBC Distribution Width
Within 2.0% Within 6.0% -
(RDW-SD)
RBC Distribution Width
Within 2.0% Within 6.0% -
(RDW-CV)
Platelet Distribution
Within 10.0% Within 20.0% -
Width (PDW)
Mean Platelet Volume
Within 4.0% Within 8.0% -
(MPV)
Platelet Large Cell
Within 15.0% Within 36.0% -
Ratio (P-LCR)
Plateletcrit Value (PCT) Within 6.0% Within 12.0% -

NRBC # Within 25.0% Within 50.0%

NRBC% Within 25.0% Within 50.0%

30.0 NEUT% or
Neutrophil Percent more WBC
Within 8.0% Within 16.0%
(NEUT%) 4.0x103/µL or
more
15.0 LYMPH% or
Lymphocyte Percent more WBC
Within 8.0% Within 16.0%
(LYMPH%) 4.0x103/µL or
more
5.0 MON % or
Monocyte Percent more WBC
Within 20.0% Within 40.0%
(MONO%) 4.0x103/µL or
more
Eosinophil Percent (EO Within 25% or +/- WBC 4.0x103/µL
Within 40.0%
%) 1.5 EO% or more
Within 40.0% or
Basophil Percent WBC 4.0x103/µL
+/-1.0 BASO% Within 50.0%
(BASO%) or more
±1.0BASO%
Neutrophil Count 1. 2 x103/µL or
Within 8.0% Within 16.0%
(NEUT#) more
Lymphocyte Count 0.6 x103/µL or
Within 8.0% Within 16.0%
(LYMPH#) more
Monocyte Count 0.2x103/µL or
Within 20.0% Within 40.0%
(MONO#) more
Within 25.0% or
+/-1.2x102µL
Eosinophil Count (EO#) Within 40.0% -
Within +/-
1.2x102/µL
 Reproducibility / Precision
Parameter Whole Blood
Capillary Mode Condition
Mode
Within 40% or
Basophil Count
Within +/- Within 50.0% -
(BASO#)
0.6x102/µL
RBC 3.0x106/µL
RET% 15% or less 35% or less
or more

RBC 3.0x106/µL
RET# 15% or less 35% or less
or more

RET-He 5% or less Not Measured

IG# 25% or less 75% or less

IG% 25% or less 75% or less

WBC-BF Within 30%

RBC-BF 40%

Linearity & Carryover

Residual or Residual ratio at specific concentration should be within the following range.

Carryover
1. Blood Cell Count (WBC, RBC, HGB, HCT, PLT)

When high value sample and control blood (High Abnormal) are used. Carryover ratio
obtained by standard analysis method should be within the following range.

2. Blood Cell Classification (NEUT#, LYMPH#, MONO#, EO#, BASO#, DIFF-WBC)

Carryover ratio or blank value after high value sample analysis should be within the
following range.
 Linearity
Parameters Whole Blood Carryover
Condition
Mode
White Blood +/-3% or Within
0-100x103/µL Within 1.0%
Cell (WBC) +/-3.0x102/µL
Red Blood Cell +/-2% or Within
0-800x104/µL Within 1.0%
(RBC) +/-3.0x104/µL
Hemoglobin
+/-2% or within
Concentration 0.0-25.0 g/dL Within 1.0%
+/-0.2 g/dL
(HGB)
Hematocrit +/-3% or within
0.0-75.0 HCT% Within 1.0%
(HCT) +/- I.0HCT%

Platelet Count +/-5% or within

0-100.0 x 104/µL Within 1.0%
(PLT) +/-1.0x104/µL
+/-10% +/-
NRBC # 2%
0.2x103/µL
NRBC % +/-20% -
RET# +/-20% -
RET% +/-20% -
+/-20% or Within RBCs < 1.0
WBC-BF 0.3%
+/-0.010x103/µL x106/µL

+/-2 % or Within
RBC-BF - 0.3%
+/-0.010x106/µL

ACCURACY
1. Blood Cell Count (WBC, RBC, PLT, Hb,)

When fresh normal blood is analyzed 10 times consecutively after instrument is

calibrated, the mean difference value from the value obtained on the standard
instrument should be within the following range.

2. Blood Cell Classification (NEUT%, LYMPH%, MONO%, EO%, BASO%)

1) When 100 or more blood samples (collected on that day) are analyzed, the
coefficient of variation should be within the following range.
2) The mean difference from the value obtained on the standard instrument
should be within the following range.

Accuracy Specification
 Accuracy
Parameters
Whole Blood Mode Pre Diluted Mode
+/-3% or within
White Blood Cell (WBC) Within +/-10%
+/-2.0x102/µL
+/-2% or within
Red Blood Cell (RBC) Within +/-8%
+/-3.0x104/µL
Hemoglobin +/- 2.0% Within +/-5%
+/-5% or within
Platelet Count (PLT) Within +/-10%
+/-1.0x104/µL

SPECIMEN REQUIREMENTS

TYPE OF SAMPLE

WHOLE BLOOD (E.D.T.A.) 3ml of whole blood at ambient temperature. Whole blood
samples for CBC should be analyzed on the same day.

Whole blood samples should be checked for clots before testing.

PATIENT PREPARATION

No preparation required. Blood specimens are collected either by venepuncture for

processing in whole mode or micro sampling by skin puncture for capillary mode
processing. For micro sampling the blood can be obtained from the ear lobe or finger of
an adult (preferable the latter) or from the heel of an infant. Ideally, large drops of blood
should exude slowly but spontaneously and only the very gentlest squeezing is
permissible. If it is necessary to squeeze firmly to obtain blood, the results are unreliable.

TYPE OF CONTAINER

EDTA vacutainer

LABELLING OF SPECIMENS

Specimens should be labeled with a unique patient identifier (Accession number) along
with name and sex of the patient mentioned on it.

SPECIMEN STORAGE & STABILITY OF WHOLE BLOOD SPECIMENS

CBC samples collected in EDTA can be obtained by venipuncture and are kept at room
temperature before analysis. It may be kept at room temperature for 4 -6 hrs.
If testing is delayed, it is stored at 2-8o C in refrigerator for 24 hrs.
After analysis samples are stored at room temperature for 4-6 hrs (for add on testing)
and after that it is stored at 2-8 o C in refrigerator for 24 hrs. Avoid hemolysed or grossly
contaminated samples.
NOTE: When the specimen is left unrefrigerated for more than 4-6 hours’ certain
changes occur within blood cells, which may produce misleading results of clinical
significance. Erythrocytes swell, the MCV increases, as does the RDW-SD. Platelets
also swell resulting in an increased MPV and P-LCR. The total WBC count may
decrease and the reliability of the electronic differential leukocyte count diminishes. The
degree of change is variable depending on the specimen and temperature at which it is
stored.
These changes are largely prevented by storage at 2-8C.

SPECIMEN TRANSPORT

Maintain and transport the specimens at room temperature.

SPECIMEN INTEGRITY

EDTA tubes for CBC should be checked upon arrival.

REAGENTS REQUIREMENT

1. Cellpack –DCL
2. Cellpack -DFL
3. Lysercell WNR
4. Lysercell WDF
5. Fluorocell WNR
6. Fluorocell WDF
7. Fluorocell RET
8. Sulfolyser
9. Cellclean Auto (4 ml) Detergent for cleaning of instrument. Storage Temp.1-300C

Reagent Storage Volume Shelf Function

Temp. life
2-350 C
Cellpack DCL 20 L 60 days Diluent, Sheath fluid

0
Cellpack DFL 2-35 C 1.5 L 60 days Diluent used in RET & PLT-F channel

Sulfolyser 1.5 L 60 days Determination of hemoglobin

1-300 C
2-350 C Lyses RBC and perforates WBC
Lysercell WNR 4L 60 days
except Basophils
0
Lysercell WDF 2-35 C 4L 90 days Lyses RBC and perforates WBC
 Fluorocell 2-350 C Stains nucleated cell for separation of
82 ml 90 days
WNR NRBC, Basophils and others WBC
Fluorocell 42 ml 90 days Stains WBC for differentiation WBC
0
WDF 2-35 C into 4 groups (Neutrophils, basophil,
Lymphocyte, Monocyte, Eosinophil)
2-350 C
Fluorocell RET 12 ml 90 days Stains reticulocytes and platelets

ENVIRONMENT AND SAFETY CONTROLS

Ref to laboratory safety manual &

MSDS sheets

CALIBRATION

Calibration is performed to compensate for any reproducible inaccuracies of the system.

If required, values are corrected by a calibration value through company engineer.

In Initial calibration by the calibrator the reference values of 10 samples are entered. The
instrument determines the calibration value automatically.

When to Calibrate

The Sysmex XN1000 needs to be calibrated:

 Before initial operation (carried out by the Sysmex service representative!);

 when quality controls show deviations in the same direction which are determined
repeatedly;
 When a major component, such as the sample rotor valve, has been replaced.
 And on yearly basis.

Running & Updation of Calibration

Calibration is done by company engineer.

Verify that the lot number is correct for the calibrator to be used.

PROCEDURAL STEPS

Analysis Procedure for daily basis

After calibration the instrument is ready for analyzing:

STARTUP PROCEDURE
Instrument Inspection:

• Check the connection of tubes and cables.

• Check if there are any bent tubes.
• Check if there is any object on top of the instrument.
• Check for any misplaced racks.
• Make sure that the devices (hub and network converters) are all powered ON.
• Discard any waste fluid in the waste container.

Reagent Inspection:

• Make sure there are extra supplies of reagents for the number of samples to
Be processed on the day of analysis. The amount of reagent needed varies with analysis
mode.

1. Make sure that the main power of the device connected to the instrument is ON.

2. Check the power for the following devices:

• Analyzer.

• Sampler.

• Display Unit.

• Printer(optional).

3. The power for the connected devices is controlled by the IPU. Hence, let the main

power switch in the „ON‟ position at all times.

4. On switching „ON‟ the instruments power, the following dialog box

appears in the IPU.
5. Enter the required information, and then click [OK]. If [Abort] is
s e l e c t ed, logon is not complete.
6. A self-test automatically runs for approximately 10 minutes

Background Check

After the temperature has stabilized, a background check is performed by repeating

background analysis for a maximum of 3 times. It performs the analysis without
aspirating the samples to verify the effect of the auto rinse. The background value is as
given below. the background check is completed.
WBC N 0.1 [X103/µL]
WBC-D 0.1 [X103/µL]
RBC 0.02 [X106/µL]
HGB 0.1 [g/dL]
PLT 10 [X103/µL]

If the background values are not at or below the acceptable value after 3 analyses, it will
be considered as background check error. For corrective measures, ref. to chapter 14 of
instrument manual.

Analyzing Samples

Performing System Analysis for Whole Blood:

Always perform a Quality Control prior to operation – before samples are

analyzed.
Check the status of the sampler unit.
Make sure sampler unit is in ready status.
Load sample racks into Right sampler pool (in front of XN).
The groove on the bottom of the rack should fit into the guide in the feeder
section.
The barcode scanner in sampler reads the barcode label.
The samples on the rack are processed by analyzers.
Remove the rack from left sampler pool after analysis completed.

Manual analysis
1. Whole Blood Mode
2. Pre diluted Mode
3. Low WBC mode

Whole Blood Mode

Check the Status indicator LED on the analyzer is in READY state (Green).
Make sure the analyzer is in READY state.
If the tube holder has not ejected out, press the mode switch to change to
manual mode.
Click the Change Analysis Mode button on the control menu.
Select Whole Blood.
Click [OK].
Click on the Manual Analysis button on the control menu.
Enter information and select discrete test.
Click [OK].
Mix the sample tube. Place the sample tube in the tube holder.
Press the start switch on the analyzer.
 Remove the sample after analysis completed.

Low WBC mode

Check the Status indicator LED on the analyzer is in READY state.

Make sure the analyzer is in READY state.
If the tube holder has not ejected out, press the mode switch.
Click the Change Analysis Mode button on the control menu. Select LWBC
mode.
Click [OK].
Click on the Manual Analysis button on the control menu.
Enter information and select discrete test.
Click [OK].
Mix the sample tube.
Place the sample tube in the tube holder.
Press the start switch on the analyzer.
Remove the sample after analysis completed.

Pre-Dilution Mode

Check the Status indicator LED on the analyzer is in READY state.

If the tube holder has not ejected out, press the mode switch.
Click the Change Analysis Mode button on the control menu. Change the
analysis mode to Pre- dilution.
Prepare sample in 1:7 dilutions. 40ul of blood is mixed with 240ul of cellpack
DCL.
Click on the Manual Analysis button on the control menu.
Enter information and select discrete test.
Click [OK].
Mix the sample tube.
Place the sample tube with open cap in the tube holder.
Press the start switch on the analyzer.
Remove the sample after analysis.

QUALITY CONTROL PROCEDURES

QUALITY ASSURANCE

This procedure is in compliance with the general AIMS Quality Assurance Guidelines
(Refer to Quality Assurance Statement).

QUALITY CONTROL

A quality control should be performed:

 Before any start of operation – prior to analyzing samples,

 At least every 12 hours during operation, if samples are > 150
 After replenishment of components,
 After maintenance,
 If there is any doubt about the accuracy of the analysis values.

CONTROL MATERIAL

XN- check (level 1) 8 x 3 mL vial

XN- check (level 2) 8x 3 mL vial
XN- check (level 3) 8 x 3 mL vial

Storage and shelf life after first opening

XN- check is to be stored at 2-8ºC before and after opening. When handled in this
manner, the unopened product has a guaranteed stability until the expiry date stated.
After opening, the product has a guaranteed stability of 7 days if returned to the
refrigerator promptly after use.

XN- check has been tested and found to provide stable parameter values after 12 hours
at room temperature (25ºC).

Additional Special Equipment

XN- check is solely intended for use with SYSMEX reagents and analyzers. If other
control materials are used the product performance of SYSMEX instruments can not be
guaranteed.

Methodology

XN- check is to be used as hematology control blood for the quality control of any
Sysmex fully-automated and semi-automated hematology analyzers. XN- check (Level
1) is for the low abnormal level, XN- check (Level 2) is for Normal, XN- check (Level 3) is
for the high abnormal level. Use of stabilized cell preparation for controlling hematology
instrumentation is an establish procedure. When handled like a patient sample and
assayed in the Quality Control mode of a properly calibrated and functioning instrument,
XN- check will provide values within the expected range indicated on the assay sheet.

Patient repeats are done in absence of control or if any parameter is still out of range
after the rerun. In absence of all three controls two to five patient repeats on the same
instrument and one repeat performed is rechecked.

QUALITY CONTROL ANALYSIS

Control samples are analyzed by the X or L-J Control programs, and the data is stored in
a quality control file. Follow the manufacturer’s instruction for handling the control
samples.
X Control analyzes the control sample twice in succession and uses the average of the
results as the control data. L-J Control uses the analysis results obtained from a single
analysis as the control data.

To display the results of QC analysis, see chapter 8 Display of QC Analysis Result”.

Quality control data is stored in the specified quality control file. Before performing
quality control with the Main Unit, see chapter 8 Quality control analysis”.

X Control
Two consecutive analyses are performed (repeat determination) and the mean of them
is compared with the expected range.

Levey-Jennings control
Only one control blood analysis is performed (detection by separate operation) and
compared with the expected range.

QC Analysis:

Follow the procedure below to perform QC analysis.

Performing QC

Remove XN check from refrigerator and equilibrate to room temperature for 15

minutes.
Mix the QC by end-to-end inversion until all red blood cells are completely re-
suspended.
Place control blood vials in a rack. Load the rack to the right sampler pool (in front of
XN).
QC auto analyze for both analyzers. Return XN check to the refrigerator. Analyse QC
results.

When quality control analysis is completed, the analysis results are displayed in the QC
Analysis dialog box.
According to the quality control method set with the IPU, either the Xbar Control dialog
box or the L-J Control dialog box appears.
For L-J control, the analysis values in the L-J Control dialog box are used.
In the QC Analysis dialog box, the data items which exceed QC control upper or lower
limits are shown with a yellow background.
Data items which exceed QC control upper or lower limits by 3 times or more are shown
with a red background.
Click Graph to display and check the newest data which was analyzed for quality control.

When control results are not within the acceptable ranges:

a. Ensure proper mixing and control material integrity, then rerun the control. If results
are still outside the acceptable ranges, do step b.
 b. Analyze a new cell control vial. If the results are still outside the acceptable ranges,
do step c.
c. Clean the system (see Diluter System) and rerun the control.
d. Review the results:

If the results are still outside the acceptable ranges, contact a Transasia representative.

If results are within the acceptable ranges, you are ready to analyze patient samples.

The control file results automatically print Auto-Print is enabled.

Displaying Levey-Jennings Control Graphs

Do this procedure to display the Levey-Jennings Control Graphs.

INTERFERENCES

INTERFERENCES IN MEASUREMENT

Interference on the Lower End of the Platelet Distribution Curve

Particles that are approximately platelet size can interfere with the platelet histogram and
count. Small particles, such as micro-bubbles, can interfere at the low end.

Microcytic Interferences on the Upper End of the Platelet Distribution Curve

Microcytic red blood cells can intrude at the upper end of the platelet distribution curve. If
the sample contains microcytes, the instrument may be able to successfully eliminate
the influence of this interference by repositioning the variable threshold and excluding
the microcytes.

BIOLOGICAL REFERENCE INTERVAL

1 2 3-6 2-6 6-12 Adult Adult

Birth Day 3 1 Year
month month month Year Year Male Female
RBC 6.0 ± 1.0 5.3 ± 1.3 4.2 ± 1.2 3.7 ± 0.6 4.7 ± 0.6 4.5 ± 0.6 4.6 ±0.6 4.6 ± 0.6 5.0 ± 0.5 4.3 ± 0.5
X 1012 /L
Hb 18 ± 4 18 ± 3 14 ± 2.5 11.2 ± 1.8 12.6 ± 1.5 12.6 ± 1.5 12.5 ± 1.5 13.5 ± 2 15.0 ± 2 13.5 ± 1.5
g /dl
PCV 0.6 ± 0.15 0.43 ± 0.1 0.35 ± 0.07 0.35 ± 0.07 0.35 ± 0.05 0.34 ± 0.04 0.37± 0.03 0.4 ± 0.05 0.45 ± 0.05 0.41 ± 0.05
L /L
MCV 110 ± 10 105 ± 13 104 ± 12 95 ± 8 75 ± 8 78 ± 6 81 ± 6 86 ± 9 92 ± 9 92 ± 9
fL
MCH 34 ± 3 34 ± 3 33 ± 3 30 ± 3 27 ± 3 37 ± 2 27 ± 3 29 ± 4 29.5 ± 2.5 29.5 ± 2.5
Pg
MCHC 330 ± 30 330 ± 40 330 ± 40 330 ± 35 330 ± 30 340 ± 20 340 ± 30 340 ± 30 330 ± 15 330 ± 15
g/L
RETIC 120 - 400 50 - 350 20 - 60 30 - 50 40 - 100 30 - 100 30 - 100 30 - 100 50 - 100 50 - 100
X 109 /L
RETIC 2 -4% 2 -4% 0.5 – 2 % 0.5 – 2 % 0.5 – 2 % 0.5 – 2 % 0.5 – 2 % 0.5 – 2 % 0.5 – 2 % 0.5 – 2 %
%
WBC 18 ± 8 15 ± 8 12 ± 7 10 ± 5 12 ± 6 11 ± 5 10 ± 5 19 ± 4 7.0 ± 3 7.0 ± 3
X 109 /L
NEUT 4 - 14 3-5 3-9 1-5 1-6 1-7 1.5 - 8 2-8
X 109 /L

NEUT% 25-75 20-75 20-75 10-45 10-45 10-45 15-70 15 - 70 40 - 80 40 - 80

LYMPH 3-8 2-8 3 - 16 4 - 10 4 - 12 3.5 - 11 6-9 1-5

X 109 /L
LYMPH 15 - 45 15 - 80 25 - 70 25 - 70 25 - 70 25 - 70 20 - 80 20 - 80 20 - 40 20 - 40
%
MONO 0.5 – 2.0 0.5 – 1.0 0.3 – 1.0 0.4 – 1.2 0.2 – 1.2 0.2 – 1.0 0.2 – 1.0 0.2 – 1.0
X 109 /L
MONO 2 - 10 2 - 10
%
EOSIN 0.1 – 1.0 0.1 – 2.0 0.2 – 1.0 0.1 – 1.0 0.1 – 1.0 0.1 – 1.0 0.1 – 1.0 0.1 – 1.0
X 109 /L
EOSIN 1-5 1-5 1-5 1-5 1-5 1-5 1-5 1-5 1-6 1-6
%
PLT 150 -450 210 -500 150 -450 210 - 650 200 - 550 200 - 550 200 -450 180 - 400 150 -400 150 -400
X 109 /L
RDW 12.8 ± 1.2 12.8 ± 1.2
(CV) %
RDW 42.5 ± 3.5 42.5 ± 3.5
(SD) fL
NRBC/
100 0-5 0-5 0-5 0-1 0-1 0-1 0-1 0-1 0-1 0-1
WBC
Ig Cells 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1 0-1
%

RESULT ANALYSIS & REVIEWING FLAGGED RESULTS AND SLIDE

REVIEW CRITERIA

Display and output of analysis Results

Automatic transmission / printing setting have been made, then graphic printing,
data printing, and host computer output are performed for the analysis results.

Data Browser
 The Data Browser displays details of the analysis results, such as sample
numerical data, scattergram and flag information. The Data Browser
displays in two ways:
 Last 20 Sample Displays the data of the last 20 samples
analyzed
 Stored Sample Displays the data of a specified sample
stored on the computer hard drive.

Data Browser Operation

Displays sample information of the analyzed sample.

Sample No. Sample Number
Pat. ID Patient ID number

Positive / Negative Display

• [Positive] Displayed if there were any abnormalities in the blood cell count or
blood cell morphology.
– [Diff.] indicates an abnormal blood cell differentiation value.
– [Morph.] indicates abnormal cell morphology.
– [Count] indicates an abnormal blood cell count.

.
• [Negative] displayed if the sample had no errors and result normal.

Error Display
 [Result] One of the following errors has occurred:
– Blood cannot be aspirated
– Insufficient blood volume
– Low count error
 [Func.] - An error other than [Result] and Barcode Reader errors has occurred.

Action Message
[Check] There may be a mix-up of samples. Otherwise, there is a significant
difference in the analysis results.
– The sample might be wrong. Check the sample.
– Significant change in XXXXX. Check the sample. (XXXXX: WBC,
HGB, MCV, PLT)
[Review] Displayed when channel difference has occurred, for example, and the
analysis results need to be reviewed.
– Difference between WNR and WDF. Check the results.
– Difference between RBC and RET. Check the results. (RBC-I and
RBC-O)
[Retest] Displays the mode and order. This is displayed to prompt an analysis.
– Reflex PLT. (When PLT-I is low reliable)
IP messages
There are 2 type of IP messages:
1. Abnormal message
- Result out of limit define by user with some exception.
2. Suspect message
 - indicate possibility that the sample is abnormal
- Abnormal morphological finding
- Abnormal histogram or scatter gram

Result / Analysis Display & Interpretation

In the event of an abnormality in the analysis data due to an analysis error, the
cause of the abnormality will be displayed as follows:
(1) “----“: Indicates that data won’t
appear due to an analysis
error.
(2) “++++”: Indicates that value exceeds
the display range.
(3) “(Blank)”: Indicates that there is no
orders.
(4) * low reliability, review slide
(5) @ result out of linearity
(6) ! value higher than back
ground check.
(7) + higher than reference interval
(8) - lower than reference interval

Q – Flag Screen

WBC
Display IP message Meaning
Blasts? Blasts? Possibility of blasts
Left Shift? Left Shift? Possibility of left shift
Atypical Ly? Atypical Lympho? Possibility of atypical
lymphocyte
Abn lympho ? Possibility of abnormal
lymphocytes
Abn Ly/Bla? Abn Lympho/Blasts? Possibility of blasts or
abnormal lymphocytes

RBC (XT-1800i)
Display IP message
RBC Agglut? RBC Agglutination? Possibility of RBC
agglutination
Turb/HGB? Turbidity/HGB Interf? Possibility of Hb
interference by
chylemia
Iron Def? Iron Deficiency? Possibility of iron
deficiency anemia
HGB Defect? HGB Defect? Possibility of Hb
abnormality
Fragments? Fragments? Possibility of
fragmented RBCs
PLT
Display IP message
 PLT Clumps? PLT Clumps? Possibility of platelet
clumps

REVIEWING FLAGGED RESULTS

Patient sample results are generated from sample analysis. There may be
instances when a patient sample result is flagged or a parameter number is
replaced by a flag.
Carefully review all patient sample results, especially results with flags
and/or messages.

POSITIVE (red backlight) A POSITIVE result indicates that the sample is

judged abnormal according to preset criteria for analysis,
numerical values and cell morphology.

NEGATIVE (green backlight) A NEGATIVE result indicates that the sample is

normal, e.g. has no analysis errors nor IP messages.

This system categorizes and flags POSITIVE results as “DIFF Abnormal”,

“MORPH Abnormal”, and/or “COUNT Abnormal” during WBC, RBC/RET, and
PLT analysis. These flags appear when abnormal cell populations are detected
during computer analysis of the particle size distributions, scattergram, and
displayed parameters.
DIFF Abnormal Indicates abnormality in WBC differential
parameters.
MORPH Abnormal Indicates abnormal cell morphology.
COUNT Abnormal Indicates abnormality in blood cell numerical count.

During normal Analysis

The IP Messages will not be displayed in the following cases:

 Quality Control Analysis Data
 Calibration Analysis Data
 Background Check Data
 Blank Data
SLIDE REVIEW CRITERIA

S. No. Parameter Primary

1 WBC <1.0 OR >30.0
2 PLT <1,50,000 OR >10,00,000
3 HGB <5g/dL or >18.5g/dL
DIFFERENTIAL
4 No diff or incomplete diff Manual
5 Neut # <1.0 or >20.0
6 Lymph# >5.0(adult) or >7.0 (<12 yrs. old)
 7 Mono# >1.5(adult) or >3.0 (<12 yrs. old)
8 Mono % >15 %
9 EOS % >20%
10 Baso% >3%
11 NRBC/ 100 WBC or % 450 nrbc/100 WBC
12 RETICS % 30%
13 Immature Granulocyte (Ig) > 10%
SUSPECT FLAGS
14 RBC fragment Flag+
15 Blast Flag Flag +
16 WBC,RBC,HGB,PLT,Rectics Lower/ higher than Lab verified
Instrument linearity

INSTRUCTION FOR DETERMINING QUANTITATIVE RESULTS WHEN A

RESULT IS NOT WITHIN THE MEASUREMENT INTERVAL

1. Pre Diluted Mode

This mode is used for analyzing a minute amount of blood collected from the `
earlobe or fingertip.
The sample is manually loaded after removing cap in tube holder and the obtained
result is automatically multiplied by 7 for reporting, which is thus comparable to the
manual mode.

2. Low WBC mode

This mode is used for analyzing samples where TLC is less than 500 cells and
no differential is given by analyzer.

LW mode increases the leukocyte measurement count by lengthening the WDF

channel measurement time (increasing the amount of specimen analyzed). In
this way, the domain in which the analyzer can perform fractionation of low-count
leukocytes, is expanded to 10 cells/μL, providing absolute neutrophilic count
(ANC) data down to low values, an important source of information in
chemotherapy and stem cell transfusions.
Here analysis volume is 174.6 µL instead of 58.2µL.

ALERT/ CRITICAL VALUES

Test Parameter Result

Hemoglobin < 7 gm% >20 gm%
TLC (cells/cc) ≤1 or ≥ 50 x 103 / µ L
ANC ≤0.5 x 103 / µ L
Platelets ≤ 20 x 103 / µ L
PCV in New Born(up to 48 hrs) > 65%
PCV in adults > 60%
Peripheral Smear Blasts > 5% , any haemoparasite, sickle cell
MAINTENANCE OF THE INSTRUMENT

Cleaning the Tube Holder

CAUTION
Risk of damage to tube holder if it is exposed to temperatures of 70°C (158°F) or
higher.
Do not heat sterilize the tube holder or subject it to temperatures of 70°C (158°F)
or higher.
Clean the tube holder with a damp cloth and distilled water. You can also use a
1% to 2%chlorine solution made from distilled water and high-quality, fragrance-
free sodium hypochlorite.

Cleaning the Outside of the Instrument

Clean the outside of the instrument with a damp cloth and distilled water to
prevent the buildup of corrosive deposits. Pay particular attention to the sampling
probe area. Clean up spills promptly.

Cleaning the Inside of the Instrument

If corrosive deposits are evident, clean the inside of the instrument with a damp
cloth and distilled water. Be careful not to wipe contaminants into the baths.

Extended Cleaning Procedure

Do this procedure to clean the baths with a 1% to 2% solution of sodium

hypochlorite:
* If you suspect a clog or fibrin. When directed by a Transasia representative.
Supplies needed:
One 5mL syringe
50mL of a 1 to 2% chlorine solution produced from high-quality, fragrance-free
sodium hypochlorite.

CLEANING PROCEDURES

a. Cleaning the Tube Holder

Clean the tube holder with a damp cloth and distilled water. You can also use a
1% to 2% chlorine solution made from distilled water and high-quality, fragrance-
free sodium hypochlorite.

A. Daily Maintenance: Shutdown

Shutting down the XN analyzer automatically:

1. Check the Status indicator LED on the analyzer.

2. Place CELLCLEAN AUTO on the 10th position of the rack.

3. Slide the groove on the rack onto the protrusion on the right side (when you face the
analyzer), and start sampler analysis.

Shutting down the XN analyzer manually:

4. Check the Status indicator LED on the analyzer.

5. Click the Analyzer menu button on the control menu.

6. Click [Shutdown].

7. Set CELLCLEAN AUTO or 4ml of cell clean into the tube holder.

8. Press the start switch on the analyzer.

9. IPU shutdowns automatically after all analyzers connected to the IPU have shut down.

10. Shutdown takes about 15minutes.

B. As needed maintenance

i. Reagent replacement

1. Click the help button on the control menu.

2. Click [Execute] to display reagent replacement screen.

Replace cellpack and Lysercell:

1. Display the [Reagent Replacement] dialog box.

2. Remove the cap from the new reagent container.

3. Input the reagent code (barcode) with handheld barcode scanner.

4. Remove the cap from the old reagent container.

5. Pull out the dispensing set straight up.

6. Insert the dispensing set straight into the new reagent container.

7. Close the cap.

8. Click [Execute].

Replace Fluorocell:

1. Display the [Reagent Replacement] dialog box.

2. Prepare the new reagent cartridge.

3. Open the top front cover.

4. Pull up the cover from the reagent that is to be replaced

5. Remove the old reagent cartridge from its holder.

6. Install the new reagent cartridge into the holder.

7. Pull down the cover on the reagent.

8. Close the top front cover.

SHUTDOWN

At the end of each day, do shutdown to rinse the instrument and place it in a standby
mode.

IMPORTANT NOTE

Highly unusual values & values due to potential analyte errors are rechecked by
performing a re assay, a review of quality control checks, instrument check for
malfunctioning and reagent for any sign of deterioration. If all test parameters meet
requirements for the tests, the results are released with appropriate comments. In
addition, the details of the same (UHID No., Patient’s Name, Referring Doctor,
parameter and reported result) are conveyed by the Section Manager / Section Head to
clinician, for critical value reporting.

If any of the parameter is not met, then appropriate measures are taken to correct the
particular defect.

POTENTIAL SOURCES OF VARIATION / NOTES AND LIMITATIONS

 Reject the specimen if clots/hemolysed/grossly contaminated samples may

give erroneous results. Request for another.

 Samples after long standing can give wrong results due to settlement of
RBCs. Previous day samples before processing the next day has to be
inverted eight times to thoroughly mix the samples. Then leave them on the
bench for one hour. After this time, process them as normal. The results and
flagging should be as good as for fresh samples.

 Lipemic Specimens. If lipemia is suspected, then the specimen is subjected

to centrifugation. The lipemic plasma is processed on the cell counter for
plasma hemoglobin. This hemoglobin is subtracted from hemoglobin
obtained from the original specimen and corrected hemoglobin is reported
along with the calculated indices.

 Presence of cold agglutinants. If red cell agglutinants are seen on smear

and in the specimen, then the EDTA specimens are incubated at 37C for
 1 to 2 hours and then process on the cell counter. If the cold agglutinants are
present, then the anomalous results (Red cell count and red cell indices) are
corrected. The results are released with comment of presence of cold
agglutinants.

 Spurious thrombocytopenia (Platelet clumps/giant platelets/ platelet satellitism)

Manual Reporting of platelet counts Whenever giant platelets or platelet clumps are
present, counter records a low platelet count. A peripheral smear examination to
countercheck the reported platelet count is carried out.

The manual platelet count is as follows:

1) Count platelets in 10 fields under oil immersion or 40x
2.) Total the no of platelets, divide by 10 and multiple by 20,000 when
counting under oil immersion / when counting under 40x multiply by 5000.
This is
the corrected platelet count.
x .
(Corrected PC = 10 X 20,000 thousands/cmm in oil immersion
( or multiply by 5000 when counting under 40x)

 Spurious thrombocytosis (RBCs fragmentation, microcytosis dust & bacterial

contamination) Manual Reporting of platelet counts when counter records an
abnormally high platelet count. A peripheral smear examination to countercheck
the reported platelet count is carried out.

 Absolute Eosinophil Count (AEC)

AEC  Eos  x WBC Count
100
Normal Range  40-440 /cmm
ALTERNATE METHODS/STORAGE SPECIMEN

In the event that all methodologies listed in this procedure are inoperable, store
specimens either as received in the department or in the same manner as a specimen
requiring verification by repeat analysis. Consult supervisory personnel for further
directions.

REFERENCES
1. XN Analyzer Manual
2. Practical Hematology – Dacie & Lewis , Editon 9