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XN-Series Flagging Interpretation Guide v2.0
XN-Series Flagging Interpretation Guide v2.0
Version 2
This document contains information that could support the user to interpret IP (Interpretive Program) messages
generated by the XN-Series analysers. IP Messages are only intended for use in the clinical laboratory and are
not for patient diagnosis. It is not intended to substitute the information available in the Instructions for Use (IFU)
and Administrator’s Guide (ADM) but as supplementary material. In case of any editorial errors or omissions
contained herein, the reference document is always the IFU and/or the ADM.
This guide was created by Sysmex Europe SE. Kindly direct your questions and comments regarding the
content to us via your local Sysmex representative.
3 Note on ‘NRBC present’ IP message and its relation to ‘WBC Abn Scattergram’ flag 13
5 Added new condition under triggers for ‘WBC Abn Scattegram’ from WDF channel 16
Deleted contents under ‘WBC Abn Scattegram’ in BF mode that were related to SW 21.12, as the
7 19
SW is no longer new
9 Adapted image depicting the two-step approach to exclude malignant samples with WDF/WPC 23
10 Added an example for the IP messages ‘Blasts?’ and ‘Abn Lympho?’ after a WPC measurement 27
11 Corrected ‘’The RBC histogram plots event size…’ to ‘’The RBC histogram plots event volume…’ 29
Inserted condition and examples related to ‘if there is no clear separation between the RBC/RET
13 36
and PLT-O clusters’ under ‘RET Abn Scattergram’
Updated text under ‘Giant Platelet?’ IP message – ‘The ‘Giant Platelet?’ flag was implemented
18 42
with SW 21.12 onwards and is part of the ‘PLT Clumps?’ flagging strategy
21 Updated ‘Possible sample interferences’ in accordance with the latest XN-Series IFU 47
The XN-Series software is designed to aid in separating specimens into POSITIVE and NEGATIVE categories
according to pre-set criteria. The system bases its judgments on comprehensive surveys of numerical data,
particle size distributions, scattergrams and provides easy-to-understand flags/messages that indicate the
instrument's findings. These flags and messages are referred to as IP messages.
A specimen is judged NEGATIVE when there are no predefined abnormalities present in the sample. The results
are generally reported without review.
The XN-Series analysers will generate a POSITIVE judgement when an IP message is present. An established
review process by lab personnel should be initiated.
POSITIVE or ERROR judgments indicate the possibility of a sample abnormality. These results should be
reviewed carefully and may require further examination in accordance with the laboratory standard operating
protocol (SOP).
Some of the action steps suggested in this guide may coincide with procedures previously established and
implemented in a lab. These action steps are merely suggested guidelines and should be used in conjunction
with the laboratory’s SOP.
2 XN Analytical Methods
Hydrodynamically focussed impedance measurement in RBC/PLT channel
◼ Volumetric measurement of red blood cells and platelets using absolute counting by DC detection method
with hydrodynamic focusing (HDF).
◼ A diluted sample is ejected from the nozzle tip and the blood cells enclosed in sheath fluid pass through a
defined path at the centre of the aperture, as depicted below.
◼ This information is plotted as a histogram, and deviations from the expected results trigger IP message(s).
Cell volume
Electric resistance
◼ The SLS haemoglobin method is based on cyanide-free reagents using sodium lauryl sulphate (SLS).
◼ The haemoglobin measurement follows the reaction steps shown in the figure below.
Flow cell
Fluorescence flow cytometry in WNR, WDF, RET, PLT-F, and WPC channels
◼ The following icons indicate that fluorescence flow cytometry is the key technology used in WNR, WDF, RET,
PLT-F, and WPC channels.
◼ Fluorescence flow cytometry (FFC) is used to analyse cells' physiological and structural properties while they
are flowing through a very narrow flow cell.
◼ First, a blood sample is aspirated and proportioned, then diluted to a pre-set ratio and labelled with a
proprietary fluorescent marker that binds specifically to nucleic acids.
◼ Next, the sample is transported into the flow cell. The sample is illuminated by a semiconductor laser beam,
which can separate the cells using three different signals, each of which provides unique information:
✓ Forward-scattered light (FSC) - cell volume.
✓ Side-scattered light (SSC) - cell content, such as the nucleus and granules.
✓ Side-fluorescence light (SFL) - the amount of nucleic acids and cell organelles.
◼ These three signals, highlighted in the image below, are used to count and differentiate WBC, nucleated RBC
(NRBC), reticulocytes, and PLT, and to detect immature and abnormal cells based on unique algorithms.
Flow cell
Semiconductor
laser
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7
3 XN-Series flagging and messages
General definitions
4
3 5
1 6
2
IP messages
Message types
◼ There are two types of IP messages, abnormal messages and suspect messages, that may be displayed for
WBC, RBC/RET, and PLT.
✓ Abnormal message: indicates that the sample is clearly abnormal. With some exceptions, the user can pre-
defined the criteria for the ‘abnormal message’ judgment.
✓ Suspect message: indicates a possibility that the sample is abnormal.
◼ [Positive] Indicates that an analysis value or cell morphology exceeds the predefined criteria for the IP
message (abnormal sample). It is displayed on a red background.
Q-Flags
The [Q-Flag] screen displays the Positive/Negative levels for suspect IP messages as a bar graph. The shown
information corresponds to the sample you selected in the [Sample Explorer] screen.
◼ The values in the bar graph range from 0 to 300, in increments of 10. Values over 100 (threshold) are
determined as positive.
◼ In addition, the following may appear in the judgment value position (when the bar graph is empty):
✓ [Discrete]: displayed in grey text. If the parameter used for the judgment was not analysed.
✓ [Error]: if the judgment was not possible.
✓ [ ]: Blank appears if a prerequisite for the judgment was not met. Also, if the suspect judgment was not
performed due to blank data, etc.
◼ The threshold for the different Q-Flags should normally never be changed (except if indicated by Sysmex
Europe SE).
◼ The value for almost every Q-Flag is semi-quantitative. It mainly reflects the degree of the specific
abnormality for more efficient screening. RBC-related flags, including ‘Turbidity/HGB Interf?’, ‘Iron
Deficiency?’ and ‘HGB Defect?’ are triggered by a single rule considering different RBC parameters.
◼ WBC-related suspect flags such as ‘Blasts/Abn Lympho? are assigned the highest score based on several
disparate rules, some of which consider cell counts while others consider ratio or cell cluster positions. On the
chronic lymphocytic leukaemia (CLL) example below, the XN analyser judged for ‘Blasts/Abn Lympho?’ within
the lymphocyte branch. In this case, the Q-flag value was calculated based on the lymphocyte count
(LYMPH#). The high ‘Blasts/Abn Lympho?’ Q-flag value of 260 is based on the elevated lymphocyte count.
◼ A ‘Blasts/Abn Lympho?’ Q-Flag of 200 does not mean double the number of blasts compared to a Q-Flag of
100 but reflects a more abnormal WDF scattergram.
◼ The ’Blasts?’ and ‘Abn Lympho?’ Q-Flag value from the WPC channel is always the same as the
corresponding Q-Flag value for ‘Blasts/Abn Lympho?’ from the WDF channel.
◼ An exception amongst the WBC-related suspect flags is the ‘Atypical Lympho?’ flag, which correlates to the
RE-LYMP% with XN-Series SW 21.12 onwards.
◼ More details on the judgement criteria underlying suspect IP messages (and the associated Q-Flags) are
individually explained in the next chapters.
* Please note that the ‘Blast or LYMPH’ area in WPC (SSC-FSC) includes abnormal lymphocytes.
◼ WBC aggregates have been detected in the WNR channel (FSCW-FSC ◼ Dashes [----] in place of the data:
scattergram, which can be viewed on the ‘User’ or ‘Lab. Only’ screens). ✓ Check if alternative data is available (e.g. WBC-D instead of WBC-N,
◼ The appearance of abnormal particles extending from the low FSC area if reliable).
in the WNR scattergram. ✓ Perform a manual differential.
◼ Asterisk [*] next to the data:
WNR scattergram (SFL-FSC) WNR scattergram (FSCW-FSC) WNR scattergram (SFL-FSC)
✓ Scan the slide for abnormal cells.
✓ If no abnormalities are found, the data with asterisks [*] can be
reported.
✓ If abnormalities are found, perform a manual differential.
◼ Asterisk [*] next to the data and ‘greyed-out’ clusters in the scattergram:
✓ Perform a manual differential.
◼ WBC count can be derived from the TNC-N count by subtracting the
WNR scattergram (SFL-FSC) WNR scattergram (SFL-FSC) NRBC count. NRBC count must be derived manually.
◼ Subtract the NRBC from the TNC according to your laboratory SOP.
SW 00-18 SW 21.12
Details:
◼ WBC and NRBC populations cannot be separated sometimes due to
highly immature NRBC, which overlap with the WBC population.
◼ WBC and NRBC#/% are masked [----] (if ‘DIFF masking’ is ON); other
WBC related parameters can also be marked/masked in XN-Series SW
versions prior to SW 22.15. With SW 22.15 and onwards, the NRBC
and relevant WBC-related parameters are masked [----] independent of
the ‘DIFF masking’ setting.
◼ Such cases are detected based on the unusual shape and position of
the WBC population, due to the inclusion of the NRBC.
Details: Details:
WBC aggregates were detected in the WNR (FSCW-FSC) scattergram Interference of particles extending from the low FSC area.
(‘User’ or ‘Lab. Only’ screen).
◼ In this case, abnormal particles are reclassified as ‘Ghost’, the low-
◼ Calculation of WBC-N is not possible due to WBC aggregates. reliability mark [*] is added to WBC-N and all WBC-related parameters
(including the WBC differential) and NRBC#/%.
◼ When the WDF channel is measured, WBC-N switches to WBC-D,
indicated by WBC &D, and WBC-N and NRBC%/# are masked [----].
Suggested actions:
Suggested actions: ◼ Check if alternative data is available (e.g. WBC-D instead of WBC-N, if
reliable).
◼ Scan the slide for NRBC.
◼ After ruling out any interference in the WDF scattergram, report
◼ If NRBC is present, perform a manual differential. If WBC-D is available, WBC-D.
manually correct the WBC-D count according to your laboratory SOP.
◼ 5-part DIFF calculation is not possible e.g. 100.0% < (LYMPH% + WDF scattergram (SSC-SFL) WDF scattergram (SSC-SFL)
MONO% + EO% + BASO% + NEUT%).
✓ Position on Y-axis.
✓ Distribution of the population.
✓ Shape (slope) of the population.
◼ Because of these anomalies, the analyser cannot identify with certainty
if the particles in that area are immature granulocytes, neutrophils, or
even unrelated interferences. SW 21.12
SW 00-18
◼ Thus, the flag ‘WBC Abn Scattergram’ is triggered, and the IG and Details:
NEUT parameters are considered unreliable [*].
◼ When the monocyte and lymphocyte populations cannot be correctly
◼ Examples of physiological triggers include an early bacterial infection differentiated, ‘WBC Abn Scattergram’ is triggered.
and sepsis/septic shock.
◼ The presence of a large ghost population could interfere with
◼ Appears together with increased NEUT-RI (corresponds to NEUT-SFL). lymphocyte and monocyte differentiation.
Low SFL signal e.g. Hb Mizuho Low SFL signal e.g. met-Hb
Details:
◼ Patient samples harbouring various unstable haemoglobin variants
typically show WDF scattergrams with a decrease in the fluorescence
signal. A similar pattern can be seen in samples with met-haemoglobin.
Details:
◼ The ‘NRBC Present’ IP message occurs when NRBC% is higher than
Interfering liposome particles in different scattergram views the predefined threshold.
Details:
◼ The threshold is user-defined and programmable; the default setting is
Some of the abnormalities judged include: 2%.
◼ WBC and ghost clusters cannot be separated. ◼ NRBC% are calculated as NRBC# / 100 WBC.
◼ Liposomal particles interfere with the WBC-BF scattergram (images ◼ Physiologically, NRBC occurs in peripheral blood only in neonates and
shown below). premature babies.
◼ Presence of cells in the high fluorescence area (HF-BF). This is an ◼ In adults and older children, the occurrence of NRBC in circulating
adjustable setting on the analyser. (For more information, please blood always indicates serious disease. NRBC are frequently seen in
contact your local Sysmex representative). the case of haemolytic anaemias, thalassaemias, systemic
haematological diseases such as MDS or leukaemias, and severe
◼ If ‘WBC Abn Scattergram’ was triggered by the ghost or other bleeding, but may also appear in a generally critical state of health, e.g.
interference of WBC in body fluid analysis, TC-BF, WBC-BF, MN#/% trauma patients from intensive care units.
and PMN#/% will be marked unreliable [*].
The ‘NRBC Present’ IP message can be switched off in the ‘Analyzer
Suggested actions:
Settings’ menu. However, if it is switched off but NRBC% exceeds the
✓ Cell count should be checked by performing a microscopy chamber threshold value, a ‘WBC Abn Scattergram’ will be triggered instead. Hence,
count. SEU recommends activating the setting.
✓ A cytospin slide should be prepared, if necessary. (For more information, please contact your local Sysmex representative.)
‘Left Shift?’
WDF scattergram (SSC-SFL) WDF scattergram (SSC-SFL) ◼ A blood smear review and the manual count are not necessary for
adults, although it would depend on the laboratory standard operating
procedure (SOP).
Details:
◼ The ‘Left Shift?’ flag indicates that the instrument has detected cells in
the region for left shift (band cells) in the WDF scattergram. When band
cells are present, they are included in the neutrophil population.
◼ An asterisk [*] appears next to the NEUT#/% and EO#/% (and IG#/%
may be marked).
◼ The flag only occurs if WBC ≧ 0.50 x103/µL.
Atypical Lymph
Abnormal Lymph
Blast
Exclusion of WBC abnormalities in most samples Separation of malignant, reactive and negative samples
No flag
‘Blasts/Abn. Lympho?’
No flag or no flag
(Negative) (Malignant or negative) ‘Blasts?’ and/or ‘Abn Lympho?’
(Malignant)
Abnormal or atypical
lymphocytes detected
Reflex to WPC
Blast gate
Blast gate positive positive in the
in the WPC WPC scattergram
scattergram
Abnormal lymphocytes
detected together with an
absolute lymphocytosis
Reflex to WPC
WNR scattergram (SFL-FSC) WDF scattergram (SSC-SFL) ◼ When blasts or abnormal lymphocytes are suspected, a smear review
should always be performed to inspect the morphology and count of the
cells.
The algorithm detects
abnormalities in the ◼ Perform and report the manual differential results according to your
lymphocyte and
monocyte clouds. Thus,
laboratory SOP.
triggering a check in the
pathological gates for
both, abnormal
lymphocytes, and
abnormal monocytes
Positive blast
gate in the
WPC
scattergram
10 0.5 – 1.1%
20 1.2 – 1.7%
30 1.8 – 2.3%
40 2.4 – 2.9%
50 3 – 3.5%
60 3.6 – 4.1%
Active phase Recovery phase Normal
70 4.2 – 4.7%
Suggested actions: 80 4.8 – 5.3%
✓ Perform a manual count and differential according to your lab SOP. 100 6 – 6.8% DEFAULT
PLT
RBC histogram RBC histogram
Small RBCs
Large RBCs
◼ The image below depicts the parameters derived from the RBC
histogram, several of which are used as judgement criteria to exclude
anomalies.
Two RBC histograms with multiple peaks
RBC histogram
100%
Frequency
LD Cell volume UD
RDW-SD
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29
‘Dimorphic Population’ Example case • Chronic Lymphocytic Leukaemia (CLL)
Details:
In this case, the small lymphocytes due to the massive
The ‘Dimorphic Population’ IP message is generated when there are leukocytosis interfere with the RBC count, as seen in the histogram.
multiple peaks in the RBC histogram pattern. Hence, the RBC count is falsely elevated. Only the first peak would
comprise the RBC, whereas the second peak is the WBC.
RBC histogram RBC histogram
WDF scattergram (SSC-SFL) WNR scattergram (SFL-FSC)
RBC histogram
◼ Dashes [----] appear in place of data for the RDW-SD and RDW-CV.
✓ marked anisocytosis
✓ multiple RBC populations
✓ RBC fragments
◼ R-MFV refers to the most frequent volume of RBC. The mean MCV and
✓ poikilocytosis mode (the most frequent volume) are equal in a normal Gaussian
✓ rouleaux or RBC agglutination (refer to suggested action for ‘RBC distribution. However, in case of outliers caused by agglutination, the
Agglutination?’ if present) mean and mode are shifted; R-MFV = mode in such a curve
◼ If no abnormalities are found, the results with the asterisk [*] may be representing the RBC population.
reported. ◼ R-MFV value can be used to manually correct the MCH, HCT and
MCHC.
NOTE: These results are derived from the Service Tab. The decision to
report these parameters is the user's responsibility.
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30
RBC suspect flags
‘Turbidity/HGB Interf?
Details:
◼ The ‘Turbidity/HGB Interf?’ IP message occurs when the MCHC is
greater than 36.5 g/dL (22.7 mmol/L) due to interference either in the
HGB measurement channel or in the RBC/PLT measurement channel;
in some cases, it is a true increase in MCHC.
◼ Asterisks [*] appear next to the HGB, MCH and MCHC parameters. The
asterisk indicates these results are unreliable.
HGB ↑
↑ MCHC =
HCT ↓
RBC Agglutination?’
Details:
◼ The ‘RBC Agglutination?’ flag is triggered when the MCHC is greater
than 40.0 g/dL (24.8 mmol/L), and certain parameters such as RBC,
MCH and the upper detection limit of the RBC histogram exceed index
values due to interference either in the HGB measurement channel or
RBC/PLT measurement channel.
◼ Asterisks [*] appear next to the RBC, RET#, HCT, MCV, MCH and
MCHC parameters. The asterisk indicates these results are unreliable.
RBC histogram
The laboratory’s challenge is to determine the cause of the increased MCHC and to take appropriate corrective action. The CBC-O concept
embedded in the Extended IPU helps to resolve the problem caused by the abovementioned interferences in traditional measurement
methodologies and automatically offers the proper corrective actions using RET channel technology (RBC-O and HGB-O). This ensures an
optimal CBC result is reported for every sample. (For more information please contact your local Sysmex representative.)
◼ The flag only occurs if RBC ≧ 0.50 x 10⁶/µL. ◼ The flag only occurs if RBC ≧ 0.50 x 10⁶/µL.
◼ The presence of this flag triggers no asterisk marks. ◼ The presence of this flag triggers no asterisk marks.
Follow-up according to your laboratory SOP for such patients. Follow-up according to your laboratory SOP for such patients.
◼ Actions may include ◼ Actions may include scanning the peripheral smear for the presence of
abnormal RBC morphology.
✓ Measure RET channel for RET-He.
✓ Scan the peripheral smear for the presence of abnormal RBC ◼ Report the presence of any clinically significant RBC morphology
morphology. abnormalities according to your laboratory SOP.
◼ This interference can lead to the analyser reporting false results, such
as a falsely increased WBC count from the WDF channel (WBC-D), as
well as a misclassified differential.
‘iRBC?’ algorithm in the WNR channel ◼ When the ‘iRBC?’ licence is activated, the analyser uses an algorithm
including FSC to separate white blood cells from the inclusion RBC, as
the interference can be clearly seen in the WDF (SSC–FSC)
◼ In the WNR channel, the RBC with inclusions do not overlap with the
scattergram. Also, in this scattergram, the interference is clearly
WBC area. Thus the WBC count from WNR (WBC-N) is not influenced
indicated by a purple colour representing the inclusion RBC.
by the infected RBC.
◼ Consequently, the WBC-D count and the differential are automatically
◼ When the ‘iRBC?’ licence is activated, the RBC with inclusions are
corrected, indicated by an ‘&’.
clearly indicated by a dark purple colour, as shown in the figure below.
✓ Scan slide for abnormal cells (RBC abnormalities, parasites etc.). Details:
◼ The ‘RET Abn Scattergram’ IP message indicates that the analyser has
(For more information on the ‘iRBC?’ flag, please contact your local detected an abnormal separation between the RBC and reticulocytes at
Sysmex representative.) RET_THR (threshold) main RET scattergram or increased activity in the
RET_UPP (RET Upper Particle Plateau) area on the RET(EXT)
scattergram.
◼ Depending on the reason for the flag, the RET%, RET#, IRF and PLT-O
parameters can be marked with an asterisk [*]. The asterisk indicates
these results are unreliable.
◼ Decisions to report with a comment or perform an alternative method ◼ Check the PLT-I count for possible interferences.
should be based on your laboratory SOP.
‘Fragments?’
Details:
◼ The ‘Fragments?’ IP message can be triggered by the RET scattergram
and/or the RBC histogram (from the RBC /PLT channel).
◼ The algorithm considers certain RBC and PLT parameters (MCV, RDW-
SD, MCHC, PLT, lower RBC discriminator*, upper PLT discriminator*)
* These are not reportable parameters but are used in the internal flagging algorithm.
RET scattergram
PLT histogram ◼ Dashes [----] may appear in place of data for the PDW, MPV, P-LCR
and PCT or they are marked with an asterisk [*], which indicates the
100% results may be unreliable.
Frequency
PLT-F channel scattergram Type ‘B’ curve’: Abnormal height at the upper
discriminator (PU).
PLT-I result is not reliable and should be checked
PLT-F scattergram (SFL-FSC) with an alternative PLT measurement method.
Such curves are typically caused due to an
interference by RBC fragments and microcytes.
*These are not reportable parameters but are used in the internal flagging algorithm.
Normal distribution of the populations in PLT-F © 2022 Sysmex Europe SE
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38
‘PLT Abn Scattergram’
If the analyser triggered ‘PLT Abn Distribution’ and the PLT count is
◼ The PLT-F channel only triggers the ‘PLT Abn Scattergram’ flag on the
marked unreliable [*]: XN-Series analyser.
◼ No RET and no PLT-F available - PLT review on the slide. ◼ This IP Message occurs when clustering in the platelet and IPF area in
◼ Scan the peripheral smear to review for the presence of abnormal RBC the PLT-F scattergram is abnormal.
or PLT morphology, such as: ◼ PLT-F, IPF# and IPF% will be marked with an [*], which indicates these
✓ Large or giant platelets results are unreliable.
✓ Small platelets
PLT-F scattergram (SFL-FSC)
✓ Platelet clumps
✓ RBC fragments
✓ Microcytic RBC
◼ Report according to your laboratory SOP if abnormal RBC, PLT, or
other morphology is noted.
NOTE: Unlike the X-Class analysers, where PLT abnormalities in the RET
channel triggered ‘PLT Abn Scattergram’, for XN, abnormalities in the PLT-
O scattergram will trigger ‘RET Abn Scattergram’ (as explained earlier)
because the ‘PLT Abn Scattergram’ IP message is exclusive for PLT-F
channel.
✓ Fibrin strands
✓ Platelet clumps
NOTE: Reviewing the feathered edge and sides of the peripheral smear is
suggested as platelet clumps, and fibrin strands may migrate to this area
during smear preparation.
◼ If platelet clumps or fibrin strands have interfered, perform one of the
following alternate procedures to obtain an accurate count:
◼ The ‘Giant Platelet?’ flag was implemented with SW 21.12 onwards and ◼ Follow your laboratory SOP.
is part of the ‘PLT Clumps?’ flagging strategy.
◼ If a slide review is performed and abnormalities are detected, report
◼ It enables the XN analyser to distinguish them from platelet clumps, results with a related comment as per your laboratory SOP.
reducing the false positive ‘PLT Clumps?’ flags by indicating the
NOTE: The availability of this flag depends on your system configuration!
possible presence of abnormally large platelets in the sample.
The analyser uses the information from the WNR (SFL-SSC) and WNR (FSCW-
FSC) scattergrams to distinguish between giant platelets and PLT clumps.
◼ [Retest] - Check the analysis mode, the order and the status of the sample, and then reanalyse.
The table below provides an overview of the different action messages, the underlying trigger and possible follow-up actions.
Channels
Action Displayed message Underlying trigger conditions Suggested Actions
involved
[Check] The sample might be Delta check is related to a previous Check the sample.
wrong. Check the measurement of the same patient. The trigger
sample. includes a floating threshold based on the
current value of certain parameters.
[Check] A significant change in Delta check is related to a previous Check the sample.
HGB. Check the measurement of the same patient. The trigger
sample. includes a floating threshold based on the
current value of certain parameters.
[Check] A significant change in Delta check is related to a previous Check the sample.
MCV. Check the measurement of the same patient. The trigger
sample. includes a floating threshold based on the
current value of certain parameters.
[Review] Difference between It is generated based on the ratio of the Total ◼ Rerun the sample.
WNR and WDF. Nucleated Count in the WDF channel (TNC-D)
◼ If the message is not eliminated, verify WBC and differential results
Check the results. to the Total Nucleated Count in the WNR
Channel (TNC-N). The ratio is calculated as according to your laboratory SOP. Possible actions may include:
TNC-D / TNC-N. ✓ Scanning the slide for abnormal cells and estimating the WBC count.
✓ Performing a manual differential if abnormal cells are observed.
NOTE: If the analyser has reported the WBC ✓ If no abnormalities are found when reviewing the smear, and the WBC
from the WDF channel, the WBC result will
estimate matches the analyser reported WBC, the results may be
have the ‘&D’ indicator appended to the
parameter. reported according to your laboratory SOP.
[Review] Aged sample? ◼ Aged samples are described most Depending on the individual workflow challenges (created by aged
samples) for each lab, the ASI may or may not be needed. If needed, it will
prominently by an increase in MCV, a
have to be activated by a Sysmex representative after obtaining a license
decrease in MCHC and changes in WDF agreement.
cloud shape and position. In case of a sample flagged as ‘Aged Sample?’, follow the laboratory SOP.
◼ These changes are not exclusive to aged Possible actions could include:
samples but occur in certain pathological ✓ If only ‘Aged Sample?’ is triggered, no smear is necessary. Re-
samples. analyse using a fresh sample.
◼ The ‘Aged Sample Identifier’ (ASI) can ✓ ‘Aged Sample? ` triggered along with a pathological flag ‘Blasts/ Abn
differentiate between aged, pathological, Lympho?’ with or without ‘Atypical Lympho?’, smear review is
and pathological samples. necessary.
[Review] Difference between It is generated based on the ratio of the PLT ◼ Rerun the sample.
PLT and PLT-F. result from the PLT-F channel and the PLT
◼ If the message is not eliminated, follow your laboratory protocol to count
Check the results. result from the impedance channel (PLT-I).
PLT-F results are always significantly higher PLT using an alternative method.
compared to PLT-I. The ratio is calculated as:
PLT-F / PLT-I.
[Retest] Reflex PLT. PLT measured by the impedance ◼ Reflex PLT-O or PLT-F. If not available, follow the laboratory protocol.
measurement method might be interfered with
by fragmented RBC, microcytes or giant
platelets leading to an unreliable PLT-I result.
◼ If the results from the initial and repeat runs are consistent, there is no
WNR scattergram (SFL-SSC) previous patient history, and the results are abnormal, confirm as
required by your laboratory SOP using smear review or an alternate
method.
◼ If the results from the initial and repeat runs are NOT consistent, consider
insufficient or non-mixing in manual mode, an overfilled tube (e.g., no air
space in the tube to enhance hand or automated mixing) or a clotted or
fibrinous sample. Reject or recollect the sample based on your laboratory
SOP.
◼ If there are any flags or unreliable results on the repeat run, follow your
laboratory SOP for that flag.
[Retest] Suspect sample, With SW 22.06 onwards, a new algorithm has ◼ Rerun the sample.
check the sample. been implemented to detect falsely high WBC-
◼ It is also possible to repeat the sample using a CBC+DIFF profile.
N caused by unknown interference.
[Retest] Confirm eosinophil From SW 22.08 onwards, when the algorithm ◼ Review the neutrophils and eosinophils in a smear review.
and neutrophil count is unsure of the NEUT/EO clusters in case of a
by other methods difficult classification, it will trigger a ‘WBC Abn
Scattergram’ message and this action
message. Both these messages are subject to
‘Service Settings’. Please speak to your local
Sysmex representative for more details.
◼ Severe red blood cell aggregation (cold agglutination) ◼ White blood cell fragments
Reticulocytes (RET) Falsely high RET count ◼ Giant platelets ◼ Malaria
◼ Possibility of PLT clumps ◼ Howell-Jolly body
10 Supporting literature
◼ Genevieve F et al. (2014): Smear microscopy revision: propositions by the GFHC, Feuillets de Biologie (Vol LVI N° 317). Link:
http://www.gfhc.fr/fr/documents/theme-3-recommandations
◼ Briggs C et al. (2012): Performance evaluation of the Sysmex haematology XN modular system. J Clin Pathol 65: 1024-30 Link:
http://dx.doi.org/10.1136/jclinpath-2012-200930 (abstract available from Sysmex upon request)
◼ Kawauchi S et al. (2013): The positions of normal leukocytes on the scattergram of the newly developed abnormal cell detection channel of the XN-
series multi-parameter automated hematology analysers. Sysmex J Int 23(1): 1-9. Link: Free online (after free registration)-
http://scientific.sysmex.co.jp/en/
◼ Berda-Haddad Y et al. (2016): Increased mean corpuscular haemoglobin concentration: artefact or pathological condition? Int J Lab Hematol
39(1):32-41 Link: https://onlinelibrary.wiley.com/doi/epdf/10.1111/ijlh.12565