Standard Operating Procedure

Subject Sysmex XS-1000i Procedure

Index Number Lab-1501
Section Laboratory
Subsection Regional Clinic/Affiliate Hospital Laboratories
Category Departmental
Contact Kuhn, Michelle L.
Last Revised 12/31/2018

References
Required document for Laboratory Accreditation by the College of American Pathologists (CAP), Centers
for Medicare and Medicaid Services (CMS) and/or COLA.

Applicable To
Employees of Gundersen Healthcare System laboratories, Gundersen St. Joseph’s Health Services
laboratories, Gundersen Tri-County Hospital laboratories, Gundersen Boscobel Area Hospital and Clinics
laboratories, and Gundersen Palmer Lutheran Hospital and Clinics laboratories and Gundersen
Moundview Hospital and clinic laboratories.

Detail
PRINCIPLE:
The System XS-1000i is a quantitative automated hematology analyzer for in-vitro diagnostic use for
determining 21 hematological parameters. Examination of the numeric and/or morphologic findings of
the complete blood count are useful in diagnosis of such disease states as anemias, leukemia’s, allergic
reactions, viral, bacterial, and parasitic infections. The Sysmex XS-1000i analyzer directly measures the
WBC, RBC, HGB, HCT, PLT, NEUT#, LYMPH#, MONO#, EO#, and BASO#. The remaining parameters are
calculated or derived: MCV, MCH, MCHC, RDW-CV, RDW-SD, MPV, and differential percentages.

The Sysmex XS-1000i counts and sizes red blood cells (RBC) and platelet (PLT) using electronic resistance
detection enhanced by hydrodynamic focusing. Hematocrit (HCT) is measured as the ratio of the total
RBC volume to whole blood using cumulative pulse height detection. Hemoglobin (HGB) is converted to
SLS-hemoglobin and read photometrically.

WBC count and differential are evaluated using flow cytometry with a semiconductor laser utilizing
scattered light and fluorescence to determine the differences in cell size, complexity and RNA/DNA
content. The WBC differential channel classifies neutrophils (NEUT), lymphocytes (LYMPH), monocytes
(MONO), eosinophils (EO), and basophils (BASO) by cellular complexity and nucleic acid content. The
differential cell placement is then enhanced utilizing Adaptive Cluster Analysis.

CLINICAL SIGNIFICANCE:
Examination of the numerical and/or morphologic findings of the complete blood count are useful in
diagnosis of such disease states such as anemias, leukemias, allergic reactions, viral, bacterial, and
parasitic infections.

The rules for flagging WBC, RBC, and platelet parameters are specific to the Gundersen Health System
Hematology laboratories and cover all resulting and reflexing contingencies for the Sysmex XS-1000i

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 Standard Operating Procedure

instrument, manual differential, and smear results. The system is interfaced with the Laboratory
Information System (LIS) for sharing patient demographics, orders, data checking, delta checks, and
results, including critical results and validation of results.

SPECIMEN:
A. The specimen – Whole blood anticoagulated with potassium EDTA is preferred.
B. Specimen Volumes required:
1. Optimal draw is a tube drawn to capacity. The collecting tube must be filled to a
minimum of one-half full for acceptable results.
2. A minimum of 1 ml whole blood is required for running specimen in the sampler or
manual mode.
3. An EDTA micro-container filled above the 250ul line is adequate for testing in the
manual mode.
C. Unacceptable specimens including those listed below must be redrawn:
1. Clotted samples or those containing clots, fibrin strands, or platelet clumps. All
microtainer specimens will be checked for clots prior to sampling by the analyzer.
All XS scattergrams/histograms will be reviewed for a platelet clump tail in the IMI
channel and/or platelet flags before results are released.
2. Grossly hemolyzed samples. This is detected in the NRBC histogram and may give a
“NRBC abnormal scatter” flag. A swirl tail pattern may also be seen in the
WBC/Baso channel.
3. Samples drawn above an IV.
D. Characteristics that may affect test results:
1. Lipemia (may increase HGB)
2. Icterus (may increase HGB)
3. Cold agglutinins (may increase WBC count; may decrease HCT & RBC count)
E. Stored Specimen Stability:
1. If stored at 4 – 8oC within 6 hours of collection, EDTA blood samples with normal
results may be analyzed up to 48 hours without significant loss of differential
stability.
2. Sample stability at room temperature is 8 hours. Samples stored at room
temperature may exhibit an increase in MCV after 24 hours. This may be minimized
by refrigeration.
3. Allow refrigerated samples to come to room temperature 30 minutes and mix by
hand inversion before analysis.
F. Do not place samples on a mechanical rocker. Constant rocking may cause PLT clumping and
alter white cell membranes resulting in false interpretive messages.

WARNING: All patients specimens should be considered potentially infectious and must be handled
with precautions used for human blood, as described in CDC recommendations and in compliance with
the Federal OSHA Bloodborne Pathogen Standard, 29 CFR part 1910.1030. Follow specimen handling as
outlined by laboratory safety policy Lab-0105 Safety: Biological.

REQUIRED: Wear gloves and lab coat. Wear safety glasses if there is a risk of splashing.

REAGENTS/MATERIALS:

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 Standard Operating Procedure

A. Supplies
1. De-ionized water
2. Kimwipes
3. Gauze, plastic lined wipes
4. Test tubes
5. CLOROX ULTRA bleach (Use when CELLCLEAN is indicated)
6. Sysmex reagents
7. Commercial controls: e-CHECK (XS) 5 x 1.5 ml vials #199-5002-0
8. Sysmex SCS-1000 whole blood calibrator
B. Sysmex Reagents
1. Four Sysmex reagents and bleach are used on the Sysmex XS-1000i.
2. All reagents are stored at room temperature and are to be used within the
manufacturer’s expiration date on each container.
3. Record date received and date opened and date expired on container. Record the lot
number, expiration date and opened date on XS-1000i reagent log.
4. All reagents are azide free, and intended for in vitro diagnostic use only; do not
ingest.
Reagent Abbreviation Open Expiration
Sysmex CELLPACK EPK 60 days
Sysmex STROMATOLYSER-4DL FFD 60 days
Sysmex STROMATOLYSER-4DS FFS 90 days
Sysmex SULFOLYSER SLS 60 days

DILUENTS:
A. Sysmex CELLPACK (EPK) is a whole blood diluent for use in the determination of hemoglobin
and impedance counting and sizing of blood cells. Sysmex CELLPACK also forms a laminar
sheath flow around the diluted sample for hydrodynamic focusing of the RBC and PLT.
Sysmex CELLPACK Active Ingredients:
Sodium Chloride 6.38 g/L
Boric Acid 1.00 g/L
Sodium Tetraborate 0.20 g/L
EDTA-2K 0.20 g/L
Sysmex CELLPACK Storage
1. Store at a controlled temperature of 5-30oC.
2. If frozen, thaw, mix thoroughly, and allow bubbles to disperse before use.
3. Sysmex CELLPACK is clear and colorless. If there are signs of contamination, instability
or color change, do not use.
Sysmex CELLPACK Stability
1. Unopened, it is stable until expiration date shown on the container.
2. Opened, Sysmex CELLPACK is stable for 60 days. Record date received and date
opened and date expired on container. Record the lot number, expiration date and
opened date on XS-1000i reagent log.
Sysmex CELLPACK Hazard Risk
The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires MSDS
documentation of ingredients which have been determined to be health hazards, comprise 1%
or greater of the composition, are physical hazards, are capable of release to exceed permissible

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 Standard Operating Procedure

exposure limit/threshold limit values or have been identified as carcinogens. Sysmex CELLPACK
does not have ingredients with those characteristics.

LYSE REAGENTS
A. Sysmex STROMATOLYSER-4DL (FFD) is the lysing reagent and diluent for the enumeration of
NEUT, LYMPH, MONO, EO, and BASO after eliminating RBC stroma.
Sysmex STROMATOLYSER-4DL Active Ingredients:
Nonionic surfactant 0.18%
Organic quaternary Ammonium salts 0.08%
Sysmex STROMATOLYSER-4DL Storage
1. Store at a controlled temperature of 2-35oC.
2. If frozen, do not use.
3. Sysmex STROMATOLYSER-4DL is a clear, odorless liquid. If there are signs of
contamination, instability or color change, do not use.
Sysmex STROMATOLYSER-4DL Stability
1. Unopened, it is stable until expiration date shown on the container.
2. Opened, Sysmex STROMATOLYSER-4DL is stable for 60 days. Record date received
and date opened and date expired on container. Record the lot number, expiration
date and opened date on XS-1000i reagent log
Sysmex STROMATOLYSER-4DL Hazard Risk
The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires MSDS
documentation of ingredients which have been determined to be health hazards, comprise 1%
or greater of the composition, are physical hazards, are capable of release to exceed permissible
exposure limit/threshold limit values or have been identified as carcinogens. Sysmex
STROMATOLYSER-4DL does not have ingredients with those characteristics.
CAUTION: Do not ingest. Avoid skin and eye contact. Flush with plenty of water immediately.
Consult with a physician in case of ingestion and/or eye contact.
B. Sysmex SULFOLYSER (SLS) is the RBC lysing reagent that releases the hemoglobin to be
measured by SLS hemoglobin method.
Sysmex SULFOLYSER Active Ingredients:
Sodium Lauryl Sulfate 1.7 g/L
Sysmex SULFOLYSER Storage
1. Store at controlled temperature of 2-30oC.
2. If frozen, may form a white cloudy precipitate. Thaw and warm reagent in a 30oC
water bath to dissolve ingredients completely and mix thoroughly before use.
3. Sysmex SULFOLYSER is a clear, odorless liquid. If there are signs of contamination,
instability or color change, do not use.
Sysmex SULFOLYSER Stability
1. Unopened, 12 months from the date of production marked on the box.
2. Opened, Sysmex SULFOLYSER is stable for 60 days. Record date received and date
opened and date expired on container. Record the lot number, expiration date and
opened date on XS-1000i reagent log.
Sysmex SULFOLYSER Health Risk
The OSHA Hazard Communication Standard of 29CFR part 1910.1200 requires MSDS
documentation of ingredients which have been determined to be health hazards, comprise 1 %
or greater of the composition, are physical hazards, are capable of release to exceed permissible

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 Standard Operating Procedure

exposure limit/threshold limit values or have been identified as carcinogens. Sysmex

SULFOLYSER does not have ingredients with those characteristics.
CAUTION: Do not ingest. Avoid skin and eye contact. Flush with plenty of water immediately.
Consult a physician in case of ingestion and/or eye contact.

STAINING REAGENTS:
A. Sysmex STROMATOLYSER-4DS (FFS) is used to stain leukocytes in diluted, lysed blood samples
for the determination of the 5-part differential including NEUT, LYMPH, MONO, EO and BASO.
Sysmex STROMATOLYSER-4DS Active Ingredients:
Ethylene Glycol 96.9%
Methanol 3.0%
Polymethine dye 0.002%
Sysmex STROMATOLYSER-4DS Storage
1. Store at controlled temperature of 3-35oC.
2. If frozen, do not use.
3. Sysmex STROMATOLYSER-4DS is a blue viscous liquid with a faint odor. If there are
signs of contamination, instability or color change, do not use.
Sysmex STROMATOLYSER-4DS Stability
1. Unopened, 12 months from the date of production marked on pouch.
2. Opened Sysmex STROMATOLYSER-4DS is stable for 90 days. Record date received,
date opened and date expired on container. Record the lot number, expiration date
and opened date on XS-1000i reagent log.
Sysmex STROMATOLYSER-4DS Health Risk
Health hazard reflects unusual exposures, via ingestion. This reagent is relatively safe when
handled while wearing gloves and when enclosed in the analyzer hydraulic system. Skin contact
may cause discoloration. Methanol component is a defatting agent, which may dry and crack
the skin. Prolonged and repeated skin contact may cause irritation.
CAUTION: Do not ingest. Avoid skin and eye contact. In the case of skin contact, wash
immediately with plenty of soap and water. In case of contact with the eyes, rinse immediately
with water or normal saline. Consult with a physician in the case of ingestion and/or eye
contact.
Recommended: Wear gloves, a lab coat and safety glasses for protection
B. CLOROXtm Germicidal Bleach
Clorox-brand bleach is recommended for use in cleaning and shutdown of the XS-1000i
analyzer whenever Cellclean is indicated. See “Maintenance” section for instructions to
prepare a 5% Clorox bleach solution as recommended for use by Sysmex.
CLOROX Ingredients:
Sodium Hypochlorite 8.25%
CLOROX Storage:
Store at room temperature (15-30oC) protected from light.
CLOROX Stability:
Stable under normal use and storage conditions.
WARNING: Avoid acidification or contact with ammonia containing products, which can
generate hazardous chlorine gas.
CLOROX Health Risk:

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 Standard Operating Procedure

WARNING: Clorox contains a strong oxidizing agent. May irritate skin. Contact with eyes can
cause severe, but temporary, injury to the eye. May cause nausea and vomiting if ingested.
Exposure to vapor or mist may irritate nose, throat and lungs.
Recommended: Wear gloves, a lab coat and safety glasses for protection.

COMMERCIAL CONTROL MATERIALS:

A. Sysmex e-CHECK and Sysmex e-CHECK (XS) are whole blood commercial controls for use with
the Sysmex XS-1000i hematology analyzer.
Sysmex e-CHECK and e-CHECK (XS) Ingredients (formulation)
Sysmex e-CHECK and e-CHECK (XS) consist of human red and white blood cells with a platelet
component suspended in fluid medium. Each vial of Sysmex e-CHECK contains 4.5 mL of
control material. Each vial of Sysmex e-CHECK (XS) contains 1.5 mL of control material.
Sysmex e-CHECK and e-CHECK (XS) Storage
1. Store vials as packaged, at 2-8oC.
2. Do not freeze or expose to excessive heat.
Sysmex e-CHECK and e-CHECK (XS) Stability
1. Unopened and properly stored, Sysmex e-CHECK and e-CHECK (XS) are stable until the
expiration date stated on the vial.
2. Open vial stability is 7 days for Sysmex e-CHECK and 14 days for Sysmex e-CHECK (XS)
when promptly refrigerated after each use.
3. Record the date on each vial upon opening.
4. Heat or freezing can damage Sysmex e-CHECK and e-CHECK (XS) without gross visible
changes. Moderate hemolysis can be normal. Deterioration is suspected when the
mean of the control results in not within the assay expected ranges after appropriate
troubleshooting.
5. If deterioration is suspected, call the Sysmex Technical Assistance Center at 1-888-
879-7639 (1-888-8SYSMEX).
WARNING: Potential Infectious Material. The human blood used in Sysmex e-CHECK/e-CHECK
(XS) is non-reactive for Hepatitis B Surface Antigen and negative for antibodies to HIV-1, HIV-2,
and Hepatitis C Virus using FDA specified techniques. However, no current tests can assure the
absence of these pathogens. Sysmex e-CHECK/e-CHECK (XS) should be considered potentially
infectious and must be handled with precautions used to human blood as described in CDC
recommendations and in compliance with the Federal OSHA Bloodborne Pathogen Standard, 29
CFR, 1910.1030.
B. Sysmex SCS-1000 Hematology Calibrator
Sysmex SCS-1000 Calibrator is a secondary whole blood calibrator for use with the Sysmex XS-
1000i hematology analyzer. Assay values for primary parameters are traceable to reference
methods.
Sysmex SCS-1000 Ingredients (formulation):
Sysmex SCS-1000 consists of human red and white blood cells with a platelet component
suspended in fluid medium. Each vial contains 2.0 mL of calibrator material.
Sysmex SCS-1000 Storage: Store vials in the upright position, at 2-8oC. Do not freeze or expose
to excessive heat.
Sysmex SCS-1000 Stability
1. Unopened and properly stored, Sysmex SCS-1000 is stable until the expiration date
stated on the vial.

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2. Open vial stability is 4 hours.

3. Storage outside of 2-8oC can damage Sysmex SCS-1000 causing deterioration that risks
inaccurate calibration. If deterioration is suspected, call the Sysmex Technical
Assistance Center at 1-888-879-7639 (1-888-8SYSMEX).
4. Use of the product at environment temperatures that exceed 86oF (30oC) can reduce
calibration accuracy.
WARNING: Potentially Infectious Material. The human blood used in Sysmex SCS-1000 is non-
reactive for Hepatitis B Surface Antigen and negative for antibodies to HIV-1, HIV-2, and Hepatitis C
Virus using FDA specified techniques. However, no current tests can assure the absence of these
pathogens. Sysmex SCS-1000 should be considered potentially infectious and must be handled with
precautions used for human blood as described in CDC recommendations and in compliance with
the Federal OSHA Bloodborne Pathogen Standard, 29 CFR, 1910. 1030.

EQUIPMENT/INSTRUMENTATION:
Sysmex XS-1000i

CALIBRATION:
Initial calibration is performed during installation. Calibration compensates for any bias inherent to the
pneumatic, hydraulic, and electrical system that may affect the accuracy of results. Calibrator traceable
to reference methods are used in the calibration of the instrument. WBC differential parameters are
calibrated in the factor prior to shipment, and verified by the field service representative upon
installation. The laboratory must verify calibration every six months or on an “as needed” basis to
ensure accuracy of the system. Service will perform the 6 month calibration and verification as part of
the Preventative Maintenance (PM).
Calibration is also required if one or more of the following occur:
1. Critical parts are re-placed such as manometers, apertures or detector circuit boards.
2. Controls show an unusual trend or are outside of acceptable limits and cannot be
corrected by maintenance or troubleshooting.
3. When advised by Sysmex Field Service Representative.

Calibration verification may include review and documentation of acceptable performance on all three
levels of commercial control, and Xm QC data, proficiency testing results and patient control testing
results. The operator may calibrate the following parameters: WBC, RBC, HGB, HCT and PLT. Before
calibration, ensure that the Sysmex XS-1000i is both clean and precise.
A. Precision Check
1. Perform routine daily and weekly maintenance on the instrument, and perform a
background count to ensure counts are within acceptable limits.
2. Verify that there is sufficient volume of all reagents. Precision and Calibration
procedures will be aborted if the Sysmex XS-1000i runs out of reagent.
3. Obtain a sample of fresh normal whole blood. Do not use commercial controls or
calibrators for precision. The blood donor specimen should:
a. be free from medication and interfering substances such as lipemia, icterus,
platelet clumps, hemolysis, ect.
b. have morphologically and numerically normal CBC.
c. be drawn in EDTA anticoagulant tube using proper collection technique.
d. be a minimum of 2 mL of sample.

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 Standard Operating Procedure

4. On the IPU, click (Menu) or press (F4).

5. Click on the “Controller” icon in the Menu window.
6. Double click on the “Precision Check” icon on the Controller menu.
7. Attach appropriate tube adapter to the sample position.
8. Place the well-mixed sample tube in the tube position.
9. Analyze the sample 11 times in the Manual Mode, mixing gently before each analysis.
After each analysis, the results display on the IPU screen, and the cursor moves to the
next line.
10. The mean, SD, and CV% are calculated on the last 10 analyses. The CV% values are
displayed in red if they exceed the Precision Limit %. If the CV% exceeds the Precision
Limits, click (CANCEL).
11. If all results are within Precision Limits, click (OK) on the Precision Check screen to
save the data and close the Precision check window.
12. To print a hard copy of the Precision results, click on “Precision check List” on the
“Controller” screen, then click (File) on the Menu Bar and click (Print).
B. Calibration Check
WARNING: Potentially Infectious Material. The human blood used in Sysmex SCS-1000 is non-
reactive for Hepatitis B Surface Antigen and negative for antibodies to HIV-1, HIV-2, and
Hepatitis C Virus using FDA specified techniques. However, no current tests can assure the
absence of these pathogens. Sysmex SCS-1000 should be considered potentially infectious and
must be handled with precautions used for human blood as described in CDC recommendations
and in compliance with the Federal OSHA Blood bone Pathogen Standard, 29 CFR, 1910. 1030.
1. Prepare the Sysmex SCS-1000 calibrator according to the product insert.
2. Click (Menu) or press (F4).
3. Click the “Controller” icon on the Main menu window.
4. Double-click the “Calibrator Calibration” icon on the Controller menu.
5. Click (ASSAY TARGET), then click in the parameter fields and use the numeric keys to
enter the target values for each parameter from the Sysmex SCS-1000 assay sheet.
Click in the next field after each entry. Click (OK) when all values have been entered.
6. Attach the appropriate sample tube adapter.
7. Remove the cap from the calibrator vial and perform 6 consecutive analyses in the
sampling mode. Do not mix in between 6 analyses. Wait until the 6th analysis is
complete, and then review results.
8. After 6 analyses, the last 5 analyses are used to calculate the Range Values and Delta
percents. The parameter values are highlighted in red if they are out of range.
a. Calibration cannot be performed if the Range Value exceeds the Max Range
Value, or if the Delta % exceeds the Service limit. Take corrective action. See
Troubleshooting section in the Sysmex XS-Series Instructions for Use manual.
b. If the Range Values and Delta Percents are within acceptable limits and
require calibration, click (ACCEPT). The parameters are displayed with a
check in the check box beside the new Compensation Rates. Parameters that
are not within acceptable limits cannot be selected for calibration.
c. Click (OK) to execute the calibration or (Cancel) if calibration is not desired or
necessary. The display returns to the Controller menu.

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9. Using another fresh vial of the calibrator, verify the calibration by repeating the
Calibration Check procedure. The analyzed values should all be within acceptable
limits. Do not execute calibration; exit the Calibration function.
10. Print the Calibration History.
a. Click (Menu) or press (F4).
b. Click the “Controller” icon.
c. Click the “Calibration History” icon.
d. Click on the calibration date in the Calibration History list.
e. To print a hard copy of the Calibration results, click on the Calibration Check
List on the “Controller” screen, then click (File) on the Menu Bar and click
(Print).
Following calibration, analyze commercial controls. Adjust target values if necessary.

MAINTENANCE:
This section includes written procedures for performing monthly and 30,000 cycle maintenance. Refer
to Sysmex XS-Series Instructions for Use manual, Chapter 9 for detailed, illustrated procedures.

WARNING for All Maintenance:

CLOROX ULTRA contains a strong oxidizing agent. Causes substantial, but temporary, eye injury. May
irritate skin. May cause nausea and vomiting if ingested. Exposure to vapor or mist may irritate nose,
throat and lungs. If contact with eyes, flush with copious amounts of water. Also, potential biohazard
exposure when performing maintenance on the Sysmex XS-1000i.

REQUIRED: Wear gloves, lab coat and safety glasses for protection.

Important Note for All Maintenance

CLOROX Germicidal Bleach is 8.25% Sodium Hypochlorite solution. The Sysmex instruction for User
manual recommends using a 5% Sodium Hypochlorite solution. Sodium Hypochlorite solutions should be
made fresh at least weekly and stored in an opaque container or in the dark to prevent deterioration.
Approximately 35 ml of a 5% solution is made fresh daily using the formula below:
(Conc. 1) x (Vol. 1) = (Conc. 2) x (Vol.2)
so (8.25%) x (Vol 1) = (5.00%) x (35 mL)
V1 = 5.00/8.25 x 35ml
V1 = 21.2 ml
Use 21.2 ml of 8.25% bleach diluted to 35 ml with distilled water to make 35 ml of 5% Sodium
Hypochlorite bleach solution.
Monthly:
1. Rinse Sequence – to be performed monthly or every 1200 cycles. Cleans the optical
detector block (Sysmex XS-Series Instructions for Use manual)
a. Click (Menu) or press (F4).
b. Click “Controller” icon on the menu screen.
c. Click “Maintenance” icon. The maintenance screen displays.
d. Click on “Monthly Rinse”. The Monthly Rinse dialog box displays.
e. Attach the appropriate sample tube adapter.
f. Place a tube of 5% Sodium hypochlorite solution (CELLCLEAN) in the tube
adapter.

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 Standard Operating Procedure

g. Press the Start switch. (Located above the sample tube position on the Main
Unit of the XS-1000i without Sampler; inside the sampler cover on the XS-
1000i with Sampler) to initiate the cleaning.
h. Record on Maintenance Log.
i. The Power Off dialog box displays when the process is complete.
j. Press (Restart) to resume operation or power off the instrument.
As-needed Maintenance
1. Waste Chamber Cleaning (Sysmex XS-Series Instruction for Use manual)
a. Click (Menu) or press (F4).
b. Click “Controller” icon on the menu screen.
c. Click “Maintenance” icon. The maintenance screen displays.
d. Click on “Rinse Waste”. The Rinse Waste dialog box displays.
e. Attach the appropriate sample tube adapter.
f. Place a tube of 5% Sodium Hypochlorite solution (CELLCLEAN) in the tube
adapter.
g. Press the Start switch. (Located above the sample tube position on the Main
Unit of the XS-1000i without Sampler; inside the sample cover on the XS-
1000i with Sampler) to initiate the cleaning.
h. The dialog box closes when the sequence is complete.
i. Record on Maintenance Log.
2. Perform Rinse Flow Cell Cleaning
Perform if Flow Cell in optical detector is suspected to be dirty. (Sysmex XS-Series
Instructions for Use manual)
a. Click (Menu) or press (F4).
b. Click “Controller” icon on the menu screen.
c. Click “Maintenance” icon. The maintenance screen displays.
d. Click on “Rinse Flow Cell”. The Rinse Flow Cell dialog box displays.
e. Attach the appropriate tube adapter.
f. Place a tube of 5% Sodium Hypochlorite solution (CELLCLEAN) in the tube
adapter.
g. Press the Start switch (Located above the sample tube position on the Main
Unit of the XS-1000i without Sampler; inside the sampler cover on the XS-
1000i with Sampler) to initiate the cleaning.
h. The dialog box closes when the sequence is complete
i. Record on Maintenance Log.
3. Perform Air Bubble Removal for Flow Cell
Perform if air bubbles in Flow Cell create abnormal aggregate pattern of scattergram.
(Sysmex XS-series Instructions for User manual)
a. Click (Menu) or press (F4).
b. Click “Controller” icon on the Menu screen.
c. Click “Maintenance” icon. The maintenance screen displays.
d. Click “Remove Air Bubbles”. The sequence begins.
e. The dialog box closes when the sequence is complete.
f. Record on Maintenance Log.
4. Remove RBC Clogs with Remove Clog Sequence

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 Standard Operating Procedure

Perform when “RBC Clog Error,” “RBC Bubble Error”, or “RBC/PLT Sampling Error”
message displays. (Sysmex XS-Series Instructions for User Manual)
a. When error message is displayed the HELP dialog box displays the error.
Click on the error.
b. Click (OK) to begin Clog Removal Sequence.
5. Alternate Method for Performing Clog Removal (when no error is displayed)
a. Click (Menu) or press (F4).
b. Click “Controller” icon on the Menu screen.
c. Click “Maintenance” icon. The maintenance screen displays.
d. Click on “Remove Clogs” icon and the sequence begins.

QUALITY CONTROL:
A. Sysmex e-CHECK (4.5 ml vials) and e-CHECK (XS) (1.5 ml vials) instructions for use.
1. Remove Sysmex e-CHECK/e-CHECK (XS) vials from refrigerator and allow them to
come to room temperature (18-25oC), for approximately 15 minutes.
2. Mix vials by gentle end-to-end inversions until the cell button in the bottom of the vial
is completely suspended.
WARNING: Potentially Infectious Material. The human blood used in Sysmex e-CHECK/e-CHECK
(XS) is non-reactive for Hepatitis B Surface Antigen and negative for antibodies to HIV-1, HIV-2,
and Hepatitis C Virus using FDA specified techniques. However, no current tests can assure the
absence of these pathogens. Sysmex e-CHECK/e-CHECK (XS) should be considered potentially
infectious and must be handled with precautions used for human blood as described in CDC
recommendations and in compliance with the Federal OSHA Blood borne Pathogen Standard, 29
CFR, 1910. 1030.
B. Frequency of Controls Use and Review
1. Sysmex e-CHECK control levels: low, normal, and high will be run on each shift.
Or
2. Sysmex e-CHECK control levels: Low and normal will be run at 0600; two levels will also
be run at 1400 and 2200 (if needed) following the predetermined schedule on the
calendar for the month.
Control results will be reviewed by the tech for each shift prior to resulting patient tests. All
repeat QC will be documented on the exception log. Normal patient control will be run at
reagent change or suspected problem. Designee will review controls and patient control.
Supervisor or technical leader will review commercial controls and patient controls monthly.
C. Entering Lot Information for a New Lot of Controls:
1. Click (QC Files) or press (F5).
2. Click on file number (1-20).
3. Click (Input) or press (F9). The QC dialog box displays.
4. Click on (down arrow) beside “Material” to select the control level.
5. Click in the field for “LOT NO.”. Using the keyboard, enter the lot number from the
vial or assay sheet.
6. Click on (down arrow) beside the “Exp. Day” to display the calendar. Click on the
expiration date in the calendar to display it in the field.
7. Click (OK) to update the lot information.
8. Repeat steps 2-7 to enter lot information for the other levels of control.
D. QC Analysis:

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 Standard Operating Procedure

1. QC analysis in the Manual mode. (XS-1000i)

a. Click (Manual) or press (F2). The Manual Sample No. dialog box opens.
b. Enter the lot number using one of the following methods:
i. Use the handheld barcode reader to scan the bar code from the vial.
ii. Use the keyboard to type the lot number.
iii. NOTE: “QC” must be upper case followed by a hyphen.
iv. Click the (QC) button on the Manual Sample No. dialog box. Click on the
QC level to be analyzed. Click on (OK).
c. Press the Open/Close switch to open the sample position.
d. Attach the appropriate tube adapter for the vial to be analyzed.
e. Place the well-mixed vial in the adapter and press the “Start” switch on the
Main Unit.
f. The QC results display when analysis is complete. Results which are outside of
acceptable limits display with red background. Click (ACCEPT) to accept the
results, or (CANCEL) to reject.
g. Follow steps a-f to analyze other levels of controls.
2. QC analysis in the Manual mode. (XS-1000i with sampler)
a. Click (Manual) or press (F2). The Manual Sample No. dialog box opens.
b. Enter the lot number by one of the following methods:
i. Use the handheld barcode reader to scan the barcode from the vial.
ii. Use the keyboard to type the lot number.
iii. NOTE: “QC” must be upper case followed by a hyphen.
iv. Click the (QC) button on the Manual Sample No. dialog box. Click on
the QC level to be analyzed. Click on (OK).
c. Attach the appropriate tube adapter for the vial to be analyzed.
d. Place the well-mixed vial in the adapter and press the “Start” switch on the
right side inside the sampler.
e. The QC results display when analysis is complete. Results which are outside of
acceptable limits display with red background. Click (ACCEPT) to accept the
results, or (CANCEL) to reject.
Follow steps a-e to analyze other levels of controls.
3. QC analysis in the Sampler mode with Barcodes. (XS-1000i with sampler)
a. Place a Sysmex rack in the rack position of the Sampler with the notch on the
rack to the right.
b. Place the well-mixed control vials in positions 8,9,10 of the Sysmex rack.
c. Attach the appropriate sample tube adapter.
d. Close the Sampler cover.
e. Click (Sampler) or press (F3).
f. The Sampler Sample No. dialog box displays. Click on the starting position for
the rack and tube position in which the vials have been placed.
g. Press the sampler Start switch on the left side of the Main Unit.
h. A dialog box displays when analysis is complete.
E. Review QC
1. Radar Charts
a. On the IPU, click (QC Files) or press (F5).
b. Click on the file to be reviewed.

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 Standard Operating Procedure

c. The Radar Chart displays beside the selected file.

d. Results of the most recent analysis display in blue. Date and time on analysis
are displayed in the Analysis Date column.
e. Results outside of acceptable limits are displayed with a red “X” and the
parameter name is displayed with a red background.
2. L-J Charts
a. On the IPU, click (QC Files) or press (F5).
b. Double click on the file to be reviewed. The Levy-Jennings chart will be
displayed.
c. Results outside of acceptable limits are displayed with a red “X” on the L-J chart.
The parameter name and the result value will be displayed with a red
background.
d. Scroll through the screens to view all parameters by using the scroll bar on the
right of the screen or press the down arrow.
Verify that all parameters fall within established limits or within the package assay range.
Contact the Sysmex Technical Assistance Center to investigate any suspected control product
failure.
F. Auto-Set Targets
Parallel test new controls by analyzing each level of control a minimum of twice a day for 5
days prior to expiration of the previous lot. After a minimum of 10 data points are
accumulated, auto-set the targets.
1. On the IPU, click (QC Files) or press (F5).
2. Double-click on the appropriate file number (1-20) to open the L-J chart.
3. Set the range of data for a target calculation by clicking on the green cursor and
dragging to the left to include all points to be used in calculation, or press (Ctrl) and (A)
to select all data.
4. Click (Input) or press (F9).
5. Click on RBC in the Target/Limit window and click and drag to include all parameters.
6. Click (Auto Setting).
7. Click the “Target” check box to select. Click (OK).
8. The targets will be auto-set to the mean of the range of points selected in the QC file
and will display in the Target/Limit window.
9. Click (OK) on the “Input Lot Information” screen.
10. Repeat steps 2-9 for each level of control.
G. Corrective Action for Out of Range QC Results.
1. If the QC sample run indicates low material volume, sampling or instrument analysis
error, log on the QC exception report and repeat the control.
2. With historical limits set, all parameters must be within range. View QC results before
running.
a. If only one level of control is out of range, repeat the control and document
on the QC exception log.
b. If multiple parameters are out of range on multiple levels of controls, check
reagents and volume and age of controls. Repeat controls. Document action
taken on the QC exception report.
c. Contact the Supervisor or Customer Support for further troubleshooting
assistance.

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 Standard Operating Procedure

H. Recording and Storage of QC Data

QC results will be stored in the analyzer during the month. Patient control results will be
stored on log sheets and reviewed by the Supervisor. QC will be reviewed by the designee. All
results will be printed and sent to the Supervisor for monthly review and stored on site.
Reagent logs will be reviewed monthly. Maintenance logs will be reviewed monthly. All
results will be stored on site.
1. Printing QC Data
a. On the IPU, click (QC Charts) or press (F5).
b. Double-click the appropriate file number (1-20) to open the L-J chart.
c. Set the range of data to be printed by clicking on the green cursor and
dragging to the left to include all points, or press (Ctrl) and (A) to select all
data.
d. Click (Out) or press (F12).
e. Click on either Report (GP) for a graphic printout of the L-J Chart of data,
statistics, targets and limits or click on Report (LP) for line print of data,
statistics, targets and limits.
f. Repeat steps b-e to print each level of QC data.
2. Save QC Data from XS Series Analyzer to Insight Desktop Folder
a. Use Insight Icon to Save QC Data.
i. Analyzer IPU Select Menu F4.
ii. Select QC Files F5.
iii. Highlight to choose lot/level.
iv. Select Insight F12.
b. Save QC to Insight Data Folder.
i. If, “Please insert disk into Drive A:” appears, click Cancel.
ii. Use arrow down key in Save in Box to select Desktop.
iii. Select Insight Data Folder.
c. Select Save
i. To re-save the same file name with additional data, highlight file. Save.
ii. Replace? Yes.
d. Problem with uploaded data.
i. Delete file(s) from folder.
ii. Make corrections on analyzer.
iii. Resave data using steps 1 through 3.
e. Repeat steps 1 through 3 to save additional lot/levels.
I. Establishing Historical LIMIT %’s for Commercial Controls
For each parameter for each level of control, an acceptable range around the mean must be
established. This range, called the ‘LIMIT %’ is based on historical performance of the
commercial control material when the instrument is in good working condition. Historical
LIMIT %s are established using three different lots of Sysmex e-CHECK/e-CHECK (XS) controls
(over a 6 month period for the 84-day dated lot). Interim Limit %s suggested by Sysmex are
used prior to establishing the analyzer-specific limits during the evaluation period. Once three
lots of QC data are collected, the CV% for each parameter is average. To establish a 3CV%
limit, multiply the average CV’s x 3. These historical limits are manually entered for the LIMIT
% in each file for each level of control and are used for all subsequent lots of controls. These

Page 14 of 26
 Standard Operating Procedure

limits should provide acceptable error detection with a low probability of false rejection, and
need not be re-established.
1. Entering QC Limit %
a. On the IPU, click (QC Files) or press (F5).
b. Click on the appropriate file (1-20).
c. Click (Input) or press (F9).
d. Each parameter for the level selected will be displayed in the “Manual Setting”
window on the right of the “Input Lot Information” screen.
e. Manually enter the LIMIT % for the first parameter in the “Limit Range %” field
and press (ENTER) on the keyboard 2 times to move to the limit for the next
parameter. Repeat for all parameters of the QC level.
f. Click (OK) to save and return to QC file list.
g. Repeat steps b-f for other levels.
J. Insight Quality Assurance Program (QAP)
Each lab is responsible for uploading the data to Sysmex Insight on the Sysmex Web Site. Data
must be submitted no later than 3 days after the due date on the Sysmex e-Check/e-Check
(XS) assay sheet.
Upload QC Data from Analyzer Desktop to Insight
1. Locate Sysmex Insight Icon and Insight Data Folder(s) on Desktop. Collecting
Open/Manual and Closed Mode QC files requires two (2) Insight data folders, (Insight
Data Manual and Insight Date Closed).
2. Open Sysmex Insight Icon and IPU Desktop.
3. Insight log-on page appears.
a. Username: Email address
b. Password: Enter Insight password
4. Facility name and primary contact appear. Select Submit QC.
5. Select type of data to be uploaded.
a. Select Data Disk Upload
6. Choose correct instrument.
a. Click on arrow key by Analyzer to choose correct analyzer. Only analyzers your
facility enrolled in the Insight program appear.
b. Shift Default All. Do not change.
c. Insert Disk into disk drive or USB Memory Stick into USB Port.
d. Click Browse.
7. Locate Insight Data folder(s) on desktop.
a. Select arrow down key in the Look in box.
b. Select desktop.
8. Click one of the three QC files. Only One QC File Can Be Uploaded At A Time.
a. The QC files for e-CHECK have the lot number followed by 0801 (level1), 0802
(level 2), and 0803 (level 3).
b. The QC files for e-CHECK (XE) have the lot number followed by the 0810 (level1),
0811 (level 2), and 0812 (level 3).
c. The QC files for e-CHECK (XS) have the lot number followed by 0804 (level),
0805 (level2), and 0806 (level 3).
d. The QC files for plt-CHECK have the lot number followed by 0660 (level 1) and
0661 (level 2).

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 Standard Operating Procedure

9. Click Open.
a. The chosen file will appear in the box before the browse button.
b. Click Submit.
10. Uploaded time, analyzer series, control, lot, level, serial number mode, and shift appear.
a. Click Submit More Files to submit more levels.
b. Repeat steps 9, 10, 11, and 12.
11. Click on Review Report to view submitted data.
12. A lot-to-date report is immediately available.
a. Close the report window by clicking the X in the upper right-hand corner. The
previous screen returns.
13. If batch is incorrect, it may be deleted at this time.
a. Click Delete next to the level to be deleted. Click Yes. Repeat process to delete
additional levels.
14. Log out.
Each lot has 2 data submission dates, approximately every 30 days for the 56-day dated
product. The submission dates are printed on the QC Assay sheet.
Submit the data by uploading to the Sysmex Insight program on the Sysmex Web site:
www.sysmex.com. Additional information may be obtained by referring to the Insight
Participant Overview Report Guide provided by the Sysmex Data Center upon
enrollment in the Insight program. Contact the Sysmex Data Center for questions
regarding data submission by telephone or e-mail.
Sysmex Insight phone: 847-996-4563
Sysmex Insight Email: insight@sysmex.com
K. Patient Moving Averages – X-barM (Xm)
1. Establishing X-barM (Xm) Historical Limit %
State how your laboratory established the Targets and Limits for Xm. Sysmex Data
Center suggests using a least 200 data points, representing 4000 samples in 20 patient
size batches. Historical Xm limits are collected over multiple reagent lots and at least 1
month, including all types of patient samples normally encountered.
2. Batch Size and Review Frequency:
Complete this section with our lab’s batch size and chart review frequency. Xm can be
monitored in lieu of a retained patient sample for a longitudinal control, if 100 or more
patients are run each day. Common batch size is 20; however, Sysmex Data Center
suggests using a larger batch size to allow about six points to be plotted per 24-hour
period. Include when and whether Xm will be turned off for specific groups of patient
specimens to avoid QC error messages related to population shifts.
a. Our batch size for Xm is 20 patient samples per batch. Each point on the Xm QC
graph represents one batch.
b. Supervisor or technical leader will review Xm charts every month.
3. Switching Xm OFF or ON
a. Click (Menu) or press (F4).
b. Click “Controller” icon.
c. Click “XbarM” icon.
d. Xm dialog box displays.
e. Click (ON) or (OFF) radio button.
f. Click (OK).

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 Standard Operating Procedure

L. Patient Control
1. Retained patient samples may be used to verify instrument performance throughout the
day or with each reagent change.
2. Select a previously run normal sample that is within the normal range (mid-range is
preferable; i.e. WBC 4.0 – 11.0; select WBC 6.0 – 9.0) for all parameters.
3. Document results on the log sheet provided.
4. Review results. Results must be within stated allowable differences. Date and initial.
5. Lab will run the patient control whenever there is a change of reagents or other
suspected problem to validate reagent quality and instrument function.
M. Establishing Limit %’s For Retained Patient Controls
1. Parameters monitored include: WBC, RBC, HGB, HCT, MCHC, MCV, PLT, RDW-SD, NEUT %,
LYMPH %.
2. The acceptable limits are:
WBC RBC HGB HCT MVC MCHC PLTS RDW-SD NEUT% LYMPH%
0.7 0.25 0.6 2.0 4.0 2.0 40.0 4.0 5 5
N. PROFICIENCY SURVEYS
1. Let samples come to room temperature.
2. Hand invert 10 times until suspended.
3. If samples are barcoded, run as a regular patient. (Cap lock must be off)
4. If samples are not barcoded, mix well and run in QC files 10, 11, and 12 in the closed
mode.
5. After all samples are analyzed, print results.
6. To print results, go to “Stored Data”.
7. Highlight the first CAP sample.
8. Press “Mark”.
9. Press “Current” for each sample.
10. Press “Cancel”.
11. Press “Output”.
12. Press “GP Graph”.
13. Clear marks by pressing “Mark” then “All Clear” then “Cancel”.
OR
1. From Main Menu choose QC File.
2. Use file numbers 11-15, one for each PT sample. DO NOT USE THE FILE LABELED OTHER 1
OR ANYTHING ELSE. Always choose # 11-15.
3. Click F9 (Input)
a. Enter the number from the sample tube (i.e. 12-1-1, 12-1-2, etc.) and click OK.
b. Click Manual, choose the file you just set up and then OK. A box should appear
just like when you are running normal QC.
c. Place sample on analyzer and push the RUN button.
d. To print results press Control, Alt, and the Print Screen Key.
e. Go to Start-Programs-Accessories-Paint
f. Click Edit and then Paste.
g. Click on File-Page Set-up-Landscape
h. Click on File and Print

Implementation

Page 17 of 26
 Standard Operating Procedure

A. Start-Up Procedure
1. Press startup on computer screen.
2. Check reagent boxes for sufficient run volume.
3. Check printer paper supply.
4. The instrument automatically performs self-check on the:
a. Microprocessor
b. Mechanical parts
c. Temperatures
d. Background count
5. Analyzer Quality Control.
B. Patient Sample Processing
1. Manual Mode – (20 uL aspirated sample volume) minimum of 500 uL in tube or 90
uL in a micro-sample container.
a. On the IPU, click (Manual) or press (F2).
b. Enter the specimen number (alpha or numeric characters) using the keyboard or
using the handheld bar code reader.
c. Click on CBC or CBC + Diff if this information is not being provided by the Host
Computer.
d. Click (OK).
e. Attach appropriate sample tube adapter.
f. Mix the patient sample 10 times by end-to-end inversion.
g. Place sample in sample tube adapter. It is not necessary to remove the cap
except when using non-pierceable micro-sample containers.
WARNING: Potential biohazard exposure when handling open patient
specimens. Follow standard precautions outlined by laboratory safety
guidelines.
Recommended: Wear gloves, a lab coat and safety glasses. Use plastic lined
gauze when opening specimen tubes.
h. Press Start switch. (Located above the sample tube position on the Main Unit of
the XS-1000i without Sampler; inside the sampler cover on the XS-1000i with
Sampler).
i. When Ready LED is lit green, repeat steps a – I for each additional sample.
2. Sampler Mode with Bar Codes – XS-1000i with Sampler (20 uL aspirated sample
volume). A minimum of 1.0 cc of blood is required in the tube for the sampler
mode.
a. Place a Sysmex rack in a rack position of the Sampler with the notch on the rack
to the right.
b. Place up to 2 racks at one time (up to 20 samples).
c. Place bar coded specimens in the rack. Ensure that labels are smooth with no
loose edges.
d. Attach the appropriate sample tube adapter.
e. Close the Sampler cover.
f. On the IPU, click (Sampler) or press (F3). The Sample number dialog box
displays.
g. Click on the starting position for the rack and tube position in which the tubes
have been placed. Press (OK).

Page 18 of 26
 Standard Operating Procedure

h. Press sampler Start switch on the left side of the Main Unit.
i. The Sysmex XS-1000i automatically mixes the sample 10 times, aspirates, and
analyzes the sample according to the bar code discrete order if bidirectional
interface is used. Results print as completed if auto-output is selected.
j. A dialog box displays when analysis is complete.
3. SHUT DOWN – Perform every 24 hours.
a. Cleans the detector and dilution lines.
i. Click (Menu) or press (F4).
ii. Double click on the “Shut down” icon.
iii. After 2 minutes, a dialog box on the IPU displays “Please power off the
analyzer”.
iv. Power off the Sysmex XS Main Unit.
v. To power off the IPU, click (File) from the menu bar, then click (Exit).
vi. Dialog box displays “Do you really want to Log off?” Click (OK).
vii. Click on Start button at the bottom of Windows desktop.
viii. Click (Shut Down). NOTE: The Restart key displays on this dialog box. If
desired, click (RESTART) to begin IPU start up process.
ix. The system displays: “Please wait while the system writes unsaved data
to the disk”.
x. Record on Maintenance Log
b. Daily shutdown procedure at the end of shift
i. Power Up Sequence
a. Press power switch on Information Processing Unit (IPU). IPU log
on must be done before powering up the Main Unit.
b. Sysmex XS-1000i program log-on displays. Log on the IPU with
your User Name (___) and press (ENTER). No password is
required. Substitute your user name and password if they have
been assigned.
ii. Press the power switch on printer.
iii. Press the power switch on right side of the Main Unit.

REAGENT REPLACEMENTS:
1. When a reagent container is empty, the alarm sounds and the Help Dialog box displays on the
IPU. “Replace Container” message with the name and abbreviation for the reagent to be
replaced is displayed.
2. Click (RESET ALARM) or press (F1) to silence the alarm. The Action area of the dialog box
displays instructions for reagent replacement.
3. Click (Execute) to open Reagent Replacement screen.
4. Once open, the Reagent Replacement screen will display “Replace” next to the reagent to be
replaced.
5. Scan the long (lower) barcode label on the container using the handheld bar code reader.
Updated information is displayed for lot number, expiration date and volume.
6. Reagent information may also be entered manually.
a. Enter the lot number and expiration date found on the reagent label.
b. Enter the number of days for “Opened Expiration”. Open dating for all reagents is 60
days.

Page 19 of 26
 Standard Operating Procedure

7. Open the new reagent container. Using clean technique to avoid contamination, remove the
line from the empty container and insert it directly into the new container. Tighten the cap on
the new container.
8. Click (Execute) to initiate priming.
9. Reagent will be aspirated to satisfy the sensor.
10. The Reagent Log will be updated once priming is complete.
11. To view the Reagent Log, click (Menu) or press (F4), click on “Controller” icon, and “Reagent
Log” icon.
12. Double click the line for the reagent that was replaced to open the Comment box. A comment
(e.g. Tech initials) may be added.
13. Run previous patient and record on retained patient log or run manufacturer control.

PROCEDURE NOTES:
A. Do not place samples on a mechanical rocker. This may alter white cell membranes and give
false interpretive messages.
B. 5% Clorox ™ bleach is recommended for using in cleaning. If Clorox is not available, generic
bleach may be used but must be filtered through a 10 micron filter. Dilute Clorox Ultra.
C. Nucleated RBCs: If 5 or more NRBC are counted on the differential, the specimen should be
sent to La Crosse for analysis or reviewed by technical leader on site.
Correct WBC count if >5 NRBCs / 100 WBC.
Corrected WBC = (WBC x 100)__
(100 + NRBC#)
Calculation example:
WBC = 13.76
NRBCs / 100 WBC = 8
Corrected WBC = 13.76 x 100 = 1376 = 12.74
100 + 8 108
If the order is a CBC, NRBCs are counted and reported with the manual differential. If the order
is a HEMG, see policy Lab-5130, Verification of XE5000-XS1000i Results, for reporting
instructions.
D. Run the EDTA tube on XS-1000i before removing the lid to make a slide.
E. Lipemic specimens generating a Turbidity/Hgb Interf?
If MCHC > 37.6:
1. Warm sample for 15-30 minutes by placing the tube in the incubator or warmer.
2. Order a RERUN by selecting the appropriate specimen ID and clicking on the
rerun/reflex icon and then clicking ok (you do not need to choose test codes).
3. Mix the warmed sample and re-run on analyzer.
4. If the warmed MCHC is < 37.5:
a. Add “Specimen warmed” to the RBC count in DI by double-clicking in the comment
box to the right of the RBC result. Click on “Insert Coded Entry” from the top
banner bar and select “Warmed Specimen” and OK. This comment now will display
next to the warmed RBC value.
b. Verify results as appropriate based on other flags present.
5. If the warmed MCHC is still > 37.6: Send sample and verification form to Gundersen
Health La Crosse lab following directions listed below.
a. If results are not needed on site and no results are entered in LIS:

Page 20 of 26
 Standard Operating Procedure

i. Reject the specimen from the DI specimen worksheet.

ii. Fill out and send a Regional Verification form (Lab-5074.4).
iii. Put sample on packing list to La Crosse Lab.
1. If results are needed on site:
i. Spin an aliquot of the sample; examine the plasma for lipemia,
hemolysis or icterus.
ii. If the plasma is moderately to grossly hemolyzed, recollect the sample.
iv. If the plasma is lipemic, icteric or if the WBC is >100,000, the HGB may
be affected. Remove the HGB, MCH and MCHC results in DI and leave
blank. Then click in the test comment column to the right of each result
field; click on “Insert Coded Entry” from the top banner bar and choose
“Unable to report due to unknown interfering substances. Sent to GL-La
Crosse lab for verification.” Click OK.
v. Send a printout of the new order to LIS department with a note to
cancel the charge for this duplicate order.
vi. Put sample on a packing list to La Crosse Lab
vii. Fill out Regional Verification form (Lab-5074.4) and send it along with
the sample to La Crosse Lab.
viii. If the plasma is clear, the RBC and HCT may be inaccurate. Do not report
the RBC, HCT, MCV or MCHC. Remove these values in DI and leave blank
Then click in the test comment column to the right of each result field;
click on “Insert Coded Entry” from the top banner bar and choose
“Unable to report due to unknown interfering substances. Sent to GL-La
Crosse lab for verification.” Click Ok. Place a new order in LIS to send to
La Crosse lab.
ix. Put sample on a packing list to La Crosse lab.
x. Fill out Regional Verification Form (Lab-5074.4), and send it along with
the sample to La Crosse lab.
F. Slides are made and stained on all “positive” flagged CBC’s. The criteria for “positive” CBC’s
are:
1. All instrument flags as identified in our DI rules
2. Children ≤ 1 month of age with an ANC <2.0
G. Differential – entire or partial vote outs of electronic differential parameters; poor separation
of WBC populations. Report manual differential.
H. Specimens for verification:
1. Follow instructions listed on the Regional Verification Form(Lab-5074.4), to send the
sample to La Crosse lab.
2. Blood specimens should be sent as soon as possible. Send (in a separate bag) the tube
of blood, your stained smear, 2 unstained smears and a copy of your analyzer printout
and the Regional verification form labeled with patient ID. Call the La Crosse lab and
let them know that a specimen is coming.
3. Please inform La Crosse if results should be called to the provider on the order form.
I. You may report below or above linearity using less than (<) or greater than (>) #. Example: if
WBC is over range, result WBC > 490 in the computer if the provider needs a result
immediately. The following specimens should be sent to La Crosse for testing following the
instructions listed on the Regional Verification Form, (Lab-5074.4):

Page 21 of 26
 Standard Operating Procedure

1. WBC greater than 490 x 103/uL.

2. RBC greater than 8.0 x 106/uL.
3. HGB greater than 24.0 g/dl.
4. Hematocrit greater than 66.0%.
5. Platelets greater than 5000 x 103/uL.
6. Specimens in which the MCHC is >37.0.
J. If RDW does not calculate due to the shape of peak and trough, use comment
“Microcytic/Macrocytic or dimorphic RBC population”.

CALCULATIONS:
Formula for manually calculating new calibration factor:
New Comp value = Current Comp value x (Ref. mean/Instrument mean)

INTERPRETATION:
Reporting Results
A. XS-1000i Reference Range:
COMPLETE BLOOD COUNT REFERENCE RANGES (6/1/2014)
WBC RBC Hgb Hct MCV MCH MCHC RDW- RDW- Plt
SD CV
Age 103/uL 106/dL g/dL % fL pg g/dL fL % 103/uL
Group
0 – 24 9.0 – 4.10 – 15.0 – 44 – 70 102 – 33 – 39 32 – 36 34 – 50 11.8 – 140 -
hours 30.0 6.70 24.0 115 15.6 385
2 – 30 5.0 – 4.10 – 15.0 – 44 – 70 102 – 33 – 39 32 – 36 34 – 50 11.8 – 140 -
days 21.0 6.70 24.0 115 15.6 385
1–5 5.0 – 3.80 – 10.5 – 32 – 42 72 – 88 24 – 30 32 – 36 24 – 50 11.8 – 140 –
months 19.5 5.40 14.0 15.6 385
6 – 24 6.0 – 3.80 – 10.5 – 32 – 42 72 – 88 24 – 30 32 – 36 34 – 50 11.8 – 140 -
months 17.5 5.04 14.0 15.6 385
2–3 6.0 – 4.00 – 11.5 – 33 – 43 76 – 90 25 – 31 32 – 36 34 – 50 11.8 – 140 -
years 17.0 5.30 14.5 15.6 385
4–5 5.5 – 4.00 – 11.5 – 33 – 43 76 – 90 25 – 31 32 – 36 34 – 50 11.8 – 140 -
years 15.5 5.30 14.5 15.6 385
6–7 5.0 – 4.00 – 11.5 – 33 – 43 76 – 90 25 – 31 32 – 36 34 – 50 11.8 – 140 -
years 14.5 5.30 14.5 15.6 385
8–9 4.5 – 4.00 – 11.5 – 33 – 43 76 – 90 25 – 31 32 – 36 34 – 50 11.8 – 140 -
years 13.5 5.30 14.5 15.6 385
10 – 15
years
Male 4.5 – 4.20 – 12.5 – 36 – 47 78 – 95 26 – 32 32 – 36 34 – 50 11.8 – 140 -
13.5 5.60 16.1 15.6 385
Female 4.5 – 4.10 – 12.0 – 35 – 45 78 – 95 26 – 32 32 – 36 34 – 50 11.8 – 140 -
13.5 5.30 15.0 15.6 385
16 – 17
years
Male 4.5 – 4.20 – 12.5 – 36 – 47 78 – 95 26 – 32 32 – 36 34 – 50 11.8 – 140 -
13.0 5.60 16.1 15.6 385

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 Standard Operating Procedure

Female 4.5 – 4.10 – 12.0 – 35 – 45 78 – 95 26 – 32 32 – 36 34 – 50 11.8 – 140 -

13.0 5.30 15.0 15.6 385
Adult
Male 3.7 – 4.50 – 13.6 – 41 – 50 82 – 99 27 – 32 32 – 36 34 – 50 11.8 – 140 -
10.4 5.80 16.7 15.6 385
Female 3.7 – 4.00 – 11.8 – 36 – 46 82 – 99 27 – 32 32 – 36 34 – 50 11.8 – 140 -
10.4 5.20 15.1 15.6 385

DIFFERENTIAL REFERENCE RANGES

ELECTRONIC DIFFERENTIAL REFERENCE RANGES – ABSOLUTE VALUES
ANC Lymph Mono EOS BASO
Age Group K/uL K/uL K/uL K/uL K/uL
0 – 24 hours 6 -26 2.30 – 10.80 0.10 – 3.60 0.00 – 2.10 0.00 – 0.60
2 – 30 days 1.5 – 10.0 2.0 – 17.0 0.10 – 3.60 0.00 – 2.10 0.00 – 0.60
1-5 months 1-9 2.10 – 13.90 0.10 – 2.30 0.00 – 1.40 0.00 – 0.40
6-11 months 1-8.5 2.80 – 13.30 0.10 – 2.10 0.00 – 1.20 0.00 – 0.40
12-23 months 1.5-8.5 2.80 – 13.30 0.10 – 2.10 0.00 – 1.20 0.00 – 0.40
2-3 years 1.5-8.5 2.60 – 12.60 0.10 – 2.00 0.00 – 1.20 0.00 – 0.30
4-5 years 1.5-8.5 1.90 – 10.10 0.10 – 1.90 0.00 – 1.10 0.00 – 0.30
6-7 years 1.5-8.0 1.20 – 7.80 0.10 – 1.70 0.00 – 1.00 0.00 – 0.30
8 – 9 years 1.5 – 8.0 1.40 – 7.60 0.10 – 1.60 0.00 – 1.00 0.00 – 0.30
10 – 11 years 1.5-8.0 1.50 – 7.60 0.10 – 1.60 0.00 – 1.00 0.00 – 0.30
12-15 years 1.8-8.0 1.30 – 6.50 0.10 – 1.60 0.00 – 1.00 0.00 – 0.30
16-17 years 1.8-8.0 1.10 – 5.90 0.10 – 1.60 0.00 – 0.90 0.00 – 0.30
Adult 2.0-8.7 0.70 – 4.70 0.00 – 1.10 0.00 – 0.50 0.0 – 0.20

B. Guidelines on Acceptable Reporting Format: Reporting Criteria and format is built into the LIS.
C. Procedures to be used in reporting critical results to physicians: Please refer to Lab-0130
Critical Call Values, Lab Reporting Protocol

LIMITATIONS:
A. XS-1000i LINEARITY:
Parameter Range Units
WBC 0.1 – 490 x103/uL
RBC 0.0 – 8.0 x106/uL
HGB 0.0 – 24.0 g/dL
HCT 0.0 – 66.0 %
PLT 0 - 5000 x103/uL
B. Known Interfering Substances: Refer to Lab-5130 Verification of XN9000/XS1000i Results for
more specific details to these items below.
1. Specimens must be free of clots and fibrin strands.
2. Marked changes in plasma constituents may cause cell to swell or shrink. The blood to
anticoagulant ratio is very important.
3. Red cell fragments or white cell cytoplasmic fragments may interfere with automated
platelet count.

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 Standard Operating Procedure

4. Cold agglutinins will produce spurious macrocytosis, elevated MCHs and MCHCs falsely
decreased RBC counts and HCTs. (Warm agglutinins and “Mixed” Warm and Cold
agglutinin will produce the same spurious results as a cold agglutinin).
5. Extremely elevated WBCs may cause turbidity and increase the hemoglobin.
6. Severely hemolyzed samples (in vitro) will falsely decrease the RBC count and
hematocrit. Abnormal histogram patterns and scatter flags should be indicators of
poor collection. Recollect hemolyzed specimens.
7. Giant platelets and clumped platelets may falsely elevate the WBC count and falsely
decrease the platelet count.
8. Megakaryocytes. When megakaryocytes are present, perform a WBC and PLT
estimate. Correct WBC count if >5 Megakaryocytes / 100 WBC.

Corrected WBC = (WBC x 100)_______

(100 + megakaryocyte#)
Calculation example:
WBC = 13.76
Megakaryocytes / 100 WBC = 8

Corrected WBC = 13.76 x 100 = 1376 = 12.74

1008 108
9. Platelet clumping and/or “platelet satellitism” can occur in specimens collected in
EDTA. This may falsely elevate the WBC and give a false decrease in the platelet count.
If the provider requests a platelet count, arrangements should be made for a citrated
specimen to be drawn and performed.
10. Abnormal paraproteins found in Multiple Myeloma patients can falsely increase the
hgb (also falsely elevated WBC and PLT counts may be seen). Send to La Crosse Lab for
correction.
11. Lipemia will falsely elevate the Hgb, MCH & MCHC.
12. Do not place samples on a mechanical rocker. Excessive mixing may induce platelet
clumping and alter white cell membranes resulting in false interpretive messages.
13. RBC fragments/microcytic RBCs may interfere with automated PLT counts.
14. Megakaryocytes may falsely increase WBC counts on automated hematology analyzer.
For trouble shooting specifics refer to the Sysmex SX 1000i Operator’s Manual.
15. Severely icteric samples may falsely elevate the Hgb value and MCH and MCHC.

DETECTION OF ERRONEOUS RESULTS:

1. The XS1000i has a bi-directional interface to the LIS. This allows the order number to print on
each instrument printout and facilitates matching paperwork to specimen and slides for manual
differential
2. An instrument IP alarm message is activated if the MCHC is greater than 36.5. We have
established an operator alert to trigger when the MCHC is greater than 37.0. This alerts the
operator to investigate a possible cold agglutinin or other interfering substance.
3. Modified Result logs are printed each morning. The Supervisor or designee will review all
reports. Corrected reports or incident reports will be initiated if necessary.
4. If the instrument cannot read the barcode label, an error number will be assigned. The operator
can edit the barcode order number in the XS IPU. The operator also has the option of rerunning

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 Standard Operating Procedure

the sample or printing new labels if necessary. All numbers and patient name will be compared
prior to verification.
5. Pending logs are monitored on a scheduled basis to detect uncompleted test and assure
adequate TAT of results.

ELECTRONIC DIFFERENTIAL TO MANUAL DIFFERENTIAL COMPARISON:

The XS1000i instrument differential is checked against a tech performed 100 cell manual differential
twice a year for correlation of results.
1. Sample selection: Select 6 whole blood samples that provide a good distribution for all cell types
classified by the instrument. Review Sysmex printout to ensure that the instrument has not
flagged any results for the CBC or Differential. Samples should be run on the analyzer and a slide
made within 2 hours of collection. Samples should be kept at room temperature during analysis.
2. Procedure:
a. The instrument’s electronic differential is identified as the reference analyzer. The
manual differential will be compared to the reference analyzer.
b. Run all samples on the reference analyzer and record results from Sysmex printout for
Neut %, Lymph %, Mono %, EOS %, and Baso % on form Lab-5074.6.
c. Perform 100 cell manual differential and record results for Neut %, Lymph %, Mono %,
EOS %, and Baso % on Regional Verification Form, (Lab-5074.4).
3. Confirm that all results for the manual differential fall within the appropriate 95% confidence
interval, as defined by the instrument’s electronic differential. See attachment Lab-5074.1 – 95%
Confidence Intervals for the defined acceptable ranges.
4. Evaluate the results, document overall acceptability, and sign off on the worksheet.
a. When completed, send form and all supporting documentation to La Crosse for
Laboratory Director review.
b. Save signed forms and supporting documentation in the appropriate log book onsite.
c. Discrepancies/questions can be addressed to the Laboratory Director, Supervisor,
Technical Leader, or Sysmex Technical Assistance Center.
d. A linear regression may be performed at the request of the medical director.
5. If corrective action is necessary, it may include:
a. Analysis of additional specimens.
b. Repeat analysis of the manual differential by the same tech.
c. Repeat analysis of the manual differential by a different tech.
d. Repeat analysis of the electronic differential.
e. Troubleshooting with Sysmex Technical Service Representatives.

REVIEW AND CHANGES:

This document and all attached forms should be reviewed optimally on an annual basis, with 2 years as
the maximum review date. Review will be done by the Technical Leader, Supervisor, Manager, Medical
Director or designated person. Changes require retyping document or form and review by the Medical
Director.

REFERENCES:
1. Sysmex XS-1000i Instructions for Use, Sysmex Corporation, Kobe, Japan, February 2006.
2. Sysmex XS-1000i Users Guide, Sysmex Corporation, Kobe, Japan, February 2006.

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 Standard Operating Procedure

3. Sysmex NE-Series User’s Guide, Sysmex Corporation (USA), Inc., Clinical Applications Division,
Los Alamitos, CA, 1991 pg. 39
4. Koepke, John. Practical Laboratory Hematology. Churchill Livingstone Inc. 1991. p. 24-25, 36-
39.
5. NCCLS. Clinical Laboratory Technical Procedures Manuals-Fourth Edition; Approved Guideline.
(GP2-A4, 2002).
6. Sysmex America Inc., Mundelein, IL. Instructions for use of Sysmex SCS-1000 with Sysmex XE-
Series Calibration Program, Rev. 11, 21-Apr-00.
7. Sysmex America, Inc., Mundelein, IL. Sysmex e-CHECK Hematology Control insert sheet for
Sysmex XE-Series and XS-1000i Analyzers, Rev. 1, 30-Aug-02.
8. Sysmex America, Inc., Mundelein, IL. Sysmex e-Check Insight Overview Guide, Appendix A- Xm
Quality Control, Version 1.0a, 14-September-00.
9. Sysmex Product Information Bulletin, Approved Cleaning Agents for Sysmex Analyzers,
Document number: 63-1033, September 2012
10. Nathan & Oski’s, Hematology of Infancy and Childhood, 7th Edition. p. 1774 Appendix 11, p. 1781
Appendix 26

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