Implementation_Manaul_XS_Series_MKT-30-1009

CAP Survey FH-9 Analysis Instructions

for XS-Series without Sampler

The following instructions and notes should be used when analyzing CAP FH-9 samples
on the X-Series Analyzers:
Notes:

• Warm control to room temperature for 15 minutes.
• Mix sample thoroughly before analysis.
• The CAP survey must be analyzed using the QC Mode and the QC barcode label

(code 128 label) which is on the vial.

MANUAL OPEN MODE: FH-9 Survey Material (CBC+DIFF)

1. With “Ready LED” green, click the [Manual] Icon or the [F2] function key on the
keyboard.

2. Enter the CAP barcoded number into the “Sample No.” field, using:
• Handheld barcode reader to scan the label.
Note: Confirm Caps Lock on the keyboard is OFF.
OR
• Keyboard to manually type the number, always type the prefix QC- before
the barcoded number (example: QC-11110805).

Note: “QC” must always be uppercase and followed with a dash.

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XS-Series Implementation Manual Section 6: CAP Survey
Document Number: MKT-30-1009 November 2008 Page 1 of 8

MANUAL OPEN MODE: FH-9 Survey Material (CBC+DIFF) –

(continued)

3. Click [OK], or press [ENTER] on the keyboard.

Note: A control entry error will display in the Error List dialog box due to the
fact that this particular “control” product is not set up in a QC file. Click
[Accept] and [Close]. This will not affect your results!

4. Select the appropriate sample tube adapter, and place it in the sample position
area of the XS-1000i with the red mark facing up. Turn the adapter clockwise
until there is a click (turn about 45°) to secure it.

5. Place the well mixed (10 times by inversion)
CAP vial into the XS sample adapter.

6. Press the white [Start] button on the right
above the sample position area.

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XS-Series Implementation Manual Section 6: CAP Survey
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MANUAL OPEN MODE: FH-9 Survey Material (CBC+DIFF) –

(continued)

7. A dialog box will display when sample measurement has completed.
8. When sample analysis has been completed, click on Explorer, deselect [Last 20],

highlight the CAP samples and print the results to the GP.

Record the results on the form provided by CAP.
9. Click “Menu” to return to Main Menu.

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XS-Series Implementation Manual Section 6: CAP Survey
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XS-Series Implementation Manual Section 6: CAP Survey
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CAP Survey FH-9 Analysis Instructions

for XS-Series with Sampler

The following instructions and notes should be used when analyzing CAP FH-9 samples
on the X-Series Analyzers:

Notes:
• Warm control to room temperature for 15 minutes.
• Mix sample thoroughly before analysis.
• The CAP survey must be analyzed using the QC Mode and the QC barcode
label (code 128 label) which is on the vial.

CLOSED MODE: FH-9 Survey Material (CBC+DIFF)

1. Click the [Controller] icon on the Main Menu.
The X-Series Controller Menu will be displayed

2. Click the [Setting] icon in the XS-Series Controller Menu.

3. Click the “Sampler Stop Conditions” tab.

4. The current sampler stop conditions will be displayed on the Sampler Stop
Conditions screen.

5. If a check mark is present in the box next to “Unregistered QC Sample”, then
click the box to remove the check mark.

6. Click [OK]. Click [Menu] to return to Main Menu.

7. Place CAP sample vials into a sampler rack in positions 6, 7, 8, 9 and 10, and
place the rack into sampler rack position 1 or 2. Rack notch must be facing right
side of instrument. If rack is already in sampler, place tubes into rack and note
rack number and tube position number.

8. Close Sampler Cover. Click the [Sampler] icon.

9. Click on the rack and tube position number in Sampler Sample No. dialog box to
initiate where the instrument will begin processing.

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XS-Series Implementation Manual Section 6: CAP Survey
Document Number: MKT-30-1009 November 2008 Page 5 of 8

CLOSED MODE: FH-9 Survey Material (CBC+DIFF) – (continued)

10. Click [OK]. Press [Start] button.

Note: A control entry error will display in the Error List dialog box due to the fact
that this particular “control” product is not set up in a QC file. Click [Accept]
and [Close]. This will not affect your results!

11. A dialog box will display when sample measurement has completed. Click [OK].

12. When sample analysis has been completed, click on Explorer, deselect [Last 20],
highlight the CAP samples and print the results to the GP.

Record the results on the form provided by CAP.

13. When all data has been recorded, follow steps 1 thru 4 to access the Sampler
Stop Conditions screen. Click the box next to “Unregistered QC Sample” to place
the check mark back in the box.

14. Click [OK]. Click [Menu] to return to Main Menu.

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XS-Series Implementation Manual Section 6: CAP Survey
Document Number: MKT-30-1009 November 2008 Page 6 of 8

MANUAL OPEN MODE: FH-9 Survey Material (CBC+DIFF)

1. With “Ready LED” green, click the [Manual] Icon or the [F2] function key on the
keyboard.

2. Enter the CAP barcoded number into the “Sample No.” field, using:
• Handheld barcode reader to scan the label.
Note: Confirm Caps Lock on the keyboard is OFF.
OR
• Keyboard to manually type the number, always type the prefix QC- before the
barcoded number (example: QC-11110805).

Note: “QC” must always be uppercase and followed with a dash.

3. Click [OK], or press [ENTER] on the keyboard.

Note: A control entry error will display in the Error List dialog box due to the fact
that this particular “control” product is not set up in a QC file. Click [Accept]
and [Close]. This will not affect your results!

4. Open the Sampler Cover.

5. Select the appropriate sample tube adapter, and place it in the sample position
area of the XS-1000i with the red mark facing up. Turn the adapter clockwise
until there is a click (turn about 45°) to secure it.

One Nelson C. White Parkway, Mundelein, IL 60060 Phone 847-996-4500 · 1-800-3SYSMEX (1-800-379-7639) www.sysmex.com

XS-Series Implementation Manual Section 6: CAP Survey
Document Number: MKT-30-1009 November 2008 Page 7 of 8

MANUAL OPEN MODE: FH-9 Survey Material (CBC+DIFF) –

(continued)

6. Place the well-mixed (10 times by inversion) CAP vial into the XS sample
adapter.

7. Press the white [Start] button on the right near the sample position inside the
sampler cover.

8. A dialog box will display when sample measurement has completed.

9. When sample analysis has been completed, click on Explorer, deselect [Last 20],
highlight the CAP samples and print the results to the GP.

Record the results on the form provided by CAP.

One Nelson C. White Parkway, Mundelein, IL 60060 Phone 847-996-4500 · 1-800-3SYSMEX (1-800-379-7639) www.sysmex.com

XS-Series Implementation Manual Section 6: CAP Survey
Document Number: MKT-30-1009 November 2008 Page 8 of 8

Validation Protocols

Reportable Range Study (Linearity)

Note: On the XS-System, the manual aspiration mode sensor has to be turned off prior to
start of running Reportable Range Studies and Carryover Study. If this sensor is not
turned off, the system will not recognize a sample with low values being aspirated.

Verification of Reportable Range
Reportable Range is measured by testing levels of an analyte that are known relative to
each other. Understanding the linear relationship of the instrument method and the
analyte concentration defines the usable assay range.

Perform reportable range studies of the directly measured parameters: WBC, RBC, HGB,
HCT, and PLT to verify the reportable range of the instrument stated in the specifications in
the XS-Series User’s Guide. Other reportable parameters are calculated from combinations
of the directly measured parameters listed in XS-Series Reportable Range Limits table.

XS-Series Reportable Range Limits

Parameter Range Units

WBC 0 - 400 x 103/µL

RBC 0 - 8.00 x 106/µL

HGB 0 - 25 g/dL

HCT 0 - 60.0 %
PLT 0 - 5000* x 103/µL

*Performed using Plasma Replacement Procedure Method during Manual Methods

Manual Method:
Analyze at least four (4), preferably five (5), concentration levels of each parameter
throughout the stated linearity range. Plot the actual measured values against the
projected recovery values and compare graphically how the plotted curve conforms to a
straight line. Linear methods have a slope of 1.0, plus or minus the within run precision
of the method and an intercept of zero.

A. Procedure for Reportable Range Studies – Verification

Note: Commercial linearity products, such as RANGE CHECK-X™, may be used
for this purpose; however, commercial products may not extend to the
highest and lowest linearity limits, and matrix effect can impact recovery on
some parameters. Refer to the product insert for any limitations.

1. Obtain or prepare samples that challenge the instrument’s linearity range.

a. Specimens found in the laboratory may have a high enough count for a
specific analyte, or may be used to create the needed “stock” sample for
dilution. Refer to Section B Preparation of “Stock” Samples for Dilution.

b. If patient specimens are used, be sure to select samples with sufficient volume
to prepare two (2) replicates at 4-5 dilution concentrations (approx. 5 ml).

c. If patient specimens are to be used to prepare a concentrated sample,
choose a specimen with sample volume > 5 ml.

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 1 of 26

A. Procedure for Reportable Range – Verification (continued)

Prepare dilutions according to the Reportable Range Dilutions table.
d. Use calibrated pipettes.
e. Avoid using serial dilutions for high end linearity as any dilution error is
magnified.
f. CELLPACK™ diluent can be used for all studies.

Reportable Range Dilutions

Tube # Vol. of Blood Vol. of Cell Pack Dil Factor Dil Conc (%)

1 1.0 ml 0 ml 1 100
.8 80
2 0.8 ml 0.2 ml .6 60
.5 50
3 0.6 ml 0.4 ml .4 40

4 0.5 ml 0.5 ml .2 20

5 0.4 ml 0.6 ml .05 5

6 0.2 ml 0.8 ml

7 0.05 ml 0.95 ml

2. Use the CBC mode to analyze samples.

3. Process each dilution in replicate.

4. Average the results from the two (2) replicates for each parameter and
concentration.

5. Calculate the expected recovery for each parameter and concentration.

Expected Recovery = (Average undiluted value) x (dilution concentration)

Example: (15.0 HGB) x (60%) = 9.0 HGB

6. Plot the expected recovery on the x-axis, the average measured result on the
y-axis. Use a separate graph for each parameter.

7. Draw the “best fit” straight line through the points and calculate the slope and
intercept of the line. Computer programs such as Microsoft® Excel can quickly
and easily plot and calculate the equation of the resulting line.

8. If the selected specimen dilutions do not extend to the lower end of linearity,
choose a sample with lower starting concentration and repeat steps 2-7 again.

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XS-Series Implementation Manual Section 7: Validation Protocols

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Reportable Range (Verification of Linearity Study) – (continued)

B. Preparation of “Stock” Samples for Dilution

1. Preparation of RBC, Hemoglobin and Hematocrit Specimens

a. Choose a patient blood sample with hemoglobin of 12-15 g/dL, with at
least 5 ml of specimen.

b. Centrifuge the specimen for five (5) minutes at high speed (maximum
packing). Remove 2/3 of the plasma.

c. Suspend the “packed cells” in the remaining plasma by inverting end to
end until there is no cell button at the bottom of the tube.

d. Analyze the concentrated sample to verify the RBC, HGB, HCT is either
at or above instrument stated linearity for each parameter.

e. Follow the Procedure for Reportable Range Studies.
f. If the selected specimen dilutions do not extend to the lower end of

linearity, choose a sample with lower hemoglobin of 8 g/dL or lower and
repeat the linearity procedure.

2. Preparation of WBC Specimens

a. Obtain three (3) or more tubes (5 ml EDTA) from the same donor with an
elevated WBC count. (50-70 x 103/µL, if possible).

b. Centrifuge all but one tube slowly (1500g) for 15 minutes.
c. Remove the buffy coat from each of the centrifuged tubes and add it to

the uncentrifuged tube.
d. Mix the tube thoroughly by inverting it end to end at least 10 times.
e. Analyze the concentrated sample to verify the WBC count is either at or

above instrument stated linearity.
f. Follow the Procedure for Linearity Studies.
g. If the selected specimen dilutions do not extend to the lower end of

linearity, choose a sample with lower WBC count of 2.0–3.0 x 103/µL or
lower and repeat the linearity procedure.

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XS-Series Implementation Manual Section 7: Validation Protocols

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Reportable Range (Verification of Linearity Study) – (continued)

3. Preparation of Platelet (PLT) Specimens

a. Obtain three (3) or more tubes (5 ml EDTA) from the same donor with an
elevated PLT count (700 x 103/µL or higher). Platelet concentrates may
be a source of platelets for linearity studies.

b. Centrifuge all but one tube slowly (1500g) for 5 minutes to yield a Platelet
Rich Plasma (PRP).

c. Remove the PRP from each tube and add it to the uncentrifuged tube.
Mix thoroughly by inverting end to end at least 10 times.

d. Analyze the concentrated sample to verify the platelet count is either at or
above instrument stated linearity.

e. Follow the Procedure for Reportable Range Studies.

f. If the selected specimen dilutions do not extend to the lower end of
linearity, choose a sample with lower PLT count (40–50 x 103/µL) and
repeat the linearity procedure.

C. Serial Dilution Method for Low End Linearity

Select a sample that is low in value but still within range of commercial linearity
material. Dilute the samples via serial dilutions. Dilutions should be made with
CELLPACK as the diluent. Since the commercial material does not reach down
to 0, another linearity study needs to be done to prove that the analyzer is linear
to 0. Continue to make and run dilutions until 0 is achieved.

Use clean technique, when transferring sample material from one tube to the
next, by using a clean pipette tip for each transfer, and wiping tip of pipette after
having aspirated the proper amount for the next serially diluted tube. Make sure
that no fluid is removed from the tip to ensure accurate dilution.

1. Label tubes 2, 4, 8, 16, 32, 64, 128, 256, etc. This number indicates the
dilution factor.

2. Place 1.0 ml of CELLPACK into each tube using a volumetric pipette.

3. Put 1.0 ml of whole blood into the tube labeled 2 (1:2 dilution).
Mix thoroughly by inversion of up to 15 times.

4. Transfer 1.0 ml of the 1:2 dilution mixture into the tube labeled 4
(1:4 dilution is made). Mix thoroughly.

5. Continue transferring dilution mixtures for all tubes.

6. If you are performing the study on more than two (2) analyzers, more than
1.0ml of sample may be needed.

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 4 of 26

C. Serial Dilution Method for Low End Linearity (continued)

7. Perform an Autorinse on the analyzer. Make sure that the background count
is as low as possible, preferably 0.

8. Turn off Blood Sensor

a. On the IPU main screen select “Settings” on the drop-down. There are
two (2) options, XS and IPU. Select ‘XS’.

b. On the left side of the Menu are options:

c. Double-click on the ‘Aspiration Sensor’ settings. This will display check
boxes on the right for the aspiration sensor.

¾ The Aspiration Sensor box is normally checked.

¾ Click the box to deselect.

¾ If an auto sampler is attached, you may also deselect for that option
as well if you wish to override the aspiration sensor when using the
sampler.

d. Click OK to return to the main screen

9. Analyze the Samples

a. Run the analyzer in LAST 20. If you run dilutions to the point of 0, the only
place that the results are available is in LAST 20, and then PRINT
SCREEN results. Remember that you need to print results intermittently,
because once results are deleted in the Last 20 Screen, they cannot be
retrieved.

b. The analyzer has a minimum count for the result to be stored in sample
explorer, therefore; when there is a low count (i.e.: linearity studies and
fluid samples), the only place to print results is from LAST 20.

c. The other option is to go into the Service Mode and print results achieved
from that menu.

d. Run in manual mode. Run the sample straight two (2) times and each
dilution two (2) times. Enter the sample number with the dilution (first
number)-aspiration number (i.e.: 1-1-1; 1-1-2; 1-2-1, 1-2-2, 1-3-1,1-3-2,
etc.). A MINIMUM OF 2 DATA POINTS (DILUTIONS) is required.
Average the data points obtained for each dilution and plot the average
result.

e. Print Screen results from the LAST 20 screen (go to file and print at top of
computer page) or from the Service Screen. If you are doing multiple
parameters at the same time, print the screen for each test (i.e.: WBCs,
PLT, etc.). You can keep diluting samples to the point where a 0 result is
reached.

f. Remember to turn blood sensor back on.

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 5 of 26

C. Serial Dilution Method for Low End Linearity (continued)

10. Enter results into appropriate linearity spreadsheet.

11. Print a report to review with customer.

Commercial products are used by the HSAS to perform Reportable Range Studies.
Alternate methods of utilizing patient samples are also acceptable to perform these
studies.

References
1. CLSI/NCCLS Evaluation of the Linearity of Quantitative Analytical Methods, Proposed

Guideline, EP6-P, Vol. 6 NO. 18
2. Sysmex Applications Bulletin R-92-116, April 15, 1992. (Printed in SJI Vol 9, No. 1)
3. Hunter, Linda, Assessing Linearity The Easy Way, MLO, June 1991, pg 33-41.
4. Plaut, David, Analytical Accuracy, MT Today, January 1994, pg 14-16.
5. Paulson, Richard; Wachtel, Mitchell, MD, Using Linearity Assessment in the Laboratory,

Laboratory Medicine, Vol 26, Number 8, August 1995, pg 526-532.

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 6 of 26

WBC Reportable Range (Linearity) Verification Studies

During the implementation of XS-Series instruments, several studies are performed in
order to introduce a new instrument into the laboratory. One of these studies is the
Reportable Range Verification Study. The XS-Series system reports one of the two (2)
WBC parameters: WBC-D when a differential is ordered and WBC-C when just a CBC is
ordered. The WBC-D and WBC-C are counted differently in the same channel, so the
question arose whether it was necessary to perform reportable range verification studies
on both parameters. This document provides comparisons of the two (2) parameters
using commercial linearity material and whole blood to show no clinically significant
difference between the two (2) WBC results; therefore, the Reportable Range
Verification Study is performed on the WBC-D parameter only.

Studies were performed to determine the feasibility of eliminating one of the WBC
linearity verification studies. Two (2) comparisons in the table below show correlation
results between the WBC-C and WBC-D parameters. One comparison used commercial
linearity material from the implementation of 33 XS-Series instruments and another
comparison used whole blood samples.

Source Number r-value Regression WBC Range
33 1.00 Equation 1.13 – 323.13
Commercial 558 1.00
Linearity Material y=1.0124x+0.0355
Whole Blood
y=1.011x-0.1161 0.03 – 271.72

Comparison between the WBC-C and WBC-D exhibited excellent correlation and
confirms that a single linearity verification study is sufficient for implementation.
Therefore, for the implementation of all XS-Series instruments, only the WBC-D linearity
verification study will be performed.

Should you have any questions regarding information, please discuss it with your HSAS
(Health System Application Specialist).

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XS-Series Implementation Manual Section 7: Validation Protocols

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Carryover Study

Carryover studies are performed to determine whether samples with high cell counts
influence subsequent samples with low cell counts. Use the study protocol below to
confirm the manufacturer claim of 1% or less carryover for WBC, RBC, HGB, HCT, and
PLT (XS User’s Guide.)

A. Samples for Carryover

1. Select samples from your laboratory population that have high values for
each parameter, or prepare samples by concentration or dilution that
approximate the high and low counts encountered in your laboratory. Refer to
Linearity Studies for suggestions on concentration methods.

Examples of High and Low Values to Use for Carryover

Parameter Example of High values Example of Low values

WBC =/> 50 x 103/ µL =/< 2.0 x 103/ µL

RBC =/> 6.00 x 106/ µL =/< 2.00 x 106/ µL

HGB =/> 18.0 g/dL =/< 6 g/dL

HCT =/> 55.0 % =/< 20 %

PLT =/> 750 x 103/ µL =/< 50 x 103/ µL

B. Procedure

1. Label samples with high values as “H”, low values with “L”.

2. Analyze the “H” sample three times consecutively. Label the last replicate “H3”.

3. Analyze the “L” sample three times consecutively. Label the first replicate
“L1”, the last “L3”.

4. Calculate the % Carryover using the formula:

L1 – L3 x 100 = % Carryover
H3 –L3

Example of RBC Carryover Calculation

First Analysis Carryover %

H1 6.41

H2 6.46

H3 6.47 2.11 - 2.09 x 100 = .45%
L1 2.11 6.47 - 2.09

L2 2.10

L3 2.09

Reference
1. Protocol for Evaluation of Automated Blood Cell Counters, International Committee for

Standardization in Hematology Publications 1986, 197-212.

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XS-Series Implementation Manual Section 7: Validation Protocols

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Correlation (HSAS assists)

Correlation is performed to determine equivalency of a new analyzer to the current
analyzer or reference method. The Sysmex XS-Series instrument correlation should
include comparison of the complete blood count and differential parameters, to the
current analyzer and or reference method. Samples analyzed for correlation may also
be used for the Clinical Sensitivity study and Reference Range verification.

A. Samples for Correlation

1. Sample Collection and Storage

a. K2EDTA or K3EDTA blood tubes filled to at least ¾ capacity.

b. Samples should be kept at room temperature during the analysis.

c. Complete blood count (CBC) samples should be analyzed within 4 hours
of collection and on both analyzers within 2 hours of each other.

d. Do not place samples on a mechanical blood mixer.

2. Sample Selection

The number of samples analyzed for correlation depends on your laboratory’s
policies and procedures for new analyzer implementation. The CLSI/NCCLS
Method Comparison and Bias Estimation Using Patient Samples, (EP9-A),
suggests at least 40 specimens. Analyzing more samples challenges the
capabilities of the new analyzer and improves the confidence of the statistics
generated from the study.

Below is a sample criterion for specimen selection for correlation and Clinical
Sensitivity studies.

a. For CBC correlation and sensitivity studies, choose 200 samples, 100
normal and 100 abnormal samples. CLSI/NCCLS H20-A suggests 200
samples to perform clinical sensitivity studies.

b. Analyze no more than 25% of the total study size on one given day.

c. “Normal” samples are those with CBC and differential parameters that are
within the reference range of your facility and judged ‘Negative’. Sources
of “normal” samples include ambulatory surgical patients, blood donors,
and employee physicals.

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XS-Series Implementation Manual Section 7: Validation Protocols

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2. Sample Selection (continued)

Below is a sample criterion for specimen selection for correlation and Clinical
Sensitivity studies.

d. “Abnormal” samples should represent blood abnormalities such as:

• Leukemias • Macrocytic Anemia

• Lymphopenia • Microcytic Anemia

• Granulocytopenia • Hemolytic Anemia

• Eosinophilia • Polycythemia

• Basophilia • Thrombocytopenia

• Lymphocytosis • Thrombocytosis

• Monocytosis • Large, Giant Platelets

• Granulocytosis • Fragmented RBCs

• Hemoglobinopathy

• Samples that challenge the extremes of the analyzer’s linearity

B. Blood Films

1. Prepare three (3) wedge-type blood films as soon as possible for each sample.

2. Label the slides with the specimen number and “A”, “B”, or “C”. “A” and “B”
slides are for each technologist performing the manual differential. The “C”
slide is for the referee should it be necessary.

C. Instrument Analysis

1. Analyze CBC blood samples within 4 hours of collection and on both the
current “reference” analyzer and the XS-Series analyzer within 2 hours of
each other.

2. Analyze the sample on the XS-Series analyzer using the discrete test order.

- CBC + Diff on the XS system to enable correlation of all parameters.

3. Organize the instrument printouts in separate folders labeled for each
analyzer. Keep printouts in like order.

Note: If the same samples are used for Clinical Sensitivity studies,
print the XS-Series Q-Flag screen and staple to the matched
instrument printouts, or record the Q-Flag numbers next to
each of the flags generated on the Graphic Printout.

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XS-Series Implementation Manual Section 7: Validation Protocols

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D. Manual Differentials using CLSI/NCCLS H20-A Standard

1. Two (2) qualified reviewers must perform the differential count and blood cell
morphology. A third reviewer is designated as a referee.

2. Each reviewer performs a 200-cell differential on each blood film using the
battlement pattern for film examination.

3. Record results on template. Utilize the electronic spreadsheet provided by
your HSAS, which will automatically average the counts to the proper
percentage.

Note: If not using the HSAS provided spreadsheet, all differential data
must be averaged and expressed to the first decimal point and
add up to 100% prior to submission.

4. Compare the two (2) 200-cell counts. If there are significant differences
between the two counts, the referee will perform a 200-cell differential.

5. The average of the two (2) 200-cell differentials is used for the differential
correlation. If the referee’s count is between the other two (2) counts, retain
the average; otherwise average the two (2) closest counts.

E. Statistical Analysis

Linear regression describes the relationship between the two methods results
and can reveal possible bias in either method. The correlation coefficient (r)
indicates whether the plotted data statistically deviates from the ideal line,
(r=1.0), and values greater than 0.95 are generally deemed satisfactory. Other
methods of comparison such as Rumke’s binomial envelope may be used when
statistical sampling errors are anticipated, manual basophil counts versus
instrument counts, for example.

1. Plot the reference method, either the current analyzer or manual method on
the x-axis and the new analyzer (XS-Series) on the y-axis.

2. Correlation of an automated differential to manual differentials of less than
400-cells results in lower “r” values, giving a false impression of poor
correlation. Poor correlation is likely due to statistical sampling errors rather
than inadequacies of the automated method.

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XS-Series Implementation Manual Section 7: Validation Protocols

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E. Statistical Analysis (continued)

3. For cell types with a low frequency of occurrence such as basophils,
eosinophils and monocytes, agreement between the two methods by
graphing results on 95% confidence curves or confidence limits tables
described by Rumke may provide better correlation than the linear regression
due to statistical sampling error.

TRUE PERCENTAGE

EXPERIMENTAL PERCENTAGE COUNT

Confidence curves (95%) for leukocyte (white blood cell) differential counts, counting
100, 200 and 500 cells. Adapted from Goldner FM Mann WN: Statistical error of
differential white count. Guys Hos Rep 88:54, 1938, with permission.

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XS-Series Implementation Manual Section 7: Validation Protocols

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Confidence Limits (95%) for Various Percentages of Blood Cells of a Given Type
as Determined by Differential Counts

n: the number of cells counted; a, the observed percentage of cells of the given type. The
limits for n=100, 200, 500 and 1,000 are exact; for n=10,000 they have been determined
with Freeman and Turkey’s approximation, as described in the Geigy tables.

From Rumke CL: The statistically expected variability in differential leukocyte counting. p. 39.
In Koepke JA (ed): Differential Leukocyte Counting. College of American Pathologists,
Skokie, IL 1978, with permission.

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XS-Series Implementation Manual Section 7: Validation Protocols

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References
1. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline.

CLSI/NCCLS Document EP9-A, December 1995, p. 3-4.

2. Reference Leukocyte Differential Count (Proportional) and Evaluation of Instrumental
Methods. CLSI/NCCLS Document H20-A, March 1992.

3. Koepke, John. Practical Laboratory Hematology. Churchill Livingstone Inc., 1991. p. 81-83,
132-135, 159-160.

4. Brown, Barbara: Hematology Principles and Procedures, Lippencott, Williams & Wilkins,
Sixth Edition, 1993. p. 28-32

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 16 of 26

Data Analysis Worksheet

Reason for Study: Evaluation Installation
Study Title: Zip:
HSAS:
Account Name: Ext: Zip:
Account Address:
Address data is to be
City: mailed to:
State:
Phone Number: City:
Account Contact: State:
Title: Phone Number:

Method 1: Serial # Downloaded Data
Method 2: Serial #
Method 3: Serial # Y or N
Method 4: Serial #
Method 5: Serial #
Method 6: Serial #
Method 7: Serial #
Method 8: Serial #

Hematology Correlation No. Samples
Hematology Reference Range No. Samples
No. Samples
Body Fluid Correlation No. Samples
Urinalysis Correlation No. Samples

Urinalysis VRI

Special Instructions/
Comments:

Prior to submission please average Differentials to first decimal point and verify they = 100%
All data will be reviewed by an HSAS prior to being sent to the customer for final review.

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HOSPITAL

ADDRESS
Instrument/SN
Correlation Study- Manual Differentials

Sample ID: 001 Sample ID: 002 Diff 2 % Average
Cell Type Diff 1 % Diff 2 % Average Cell Type Diff 1 %

PMN (Seg) PMN (Seg)
Band Band
Lymph Lymph
Mono Mono
Eos Eos
Baso Baso
Atyp Lym Atyp Lym
Meta Meta
Myelo Myelo
Promyelo Promyelo
Blast Blast
NRBC NRBC

Sample ID: 003 Diff 2 % Average Sample ID: 004 Diff 2 % Average
Cell Type Diff 1 % Cell Type Diff 1 %

PMN (Seg) PMN (Seg)
Band Band
Lymph Lymph
Mono Mono
Eos Eos
Baso Baso
Atyp Lym Atyp Lym
Meta Meta
Myelo Myelo
Promyelo Promyelo
Blast Blast
NRBC NRBC

Sample ID: 005 Diff 2 % Average Sample ID: 006 Diff 2 % Average
Cell Type Diff 1 % Cell Type Diff 1 %

PMN (Seg) PMN (Seg)
Band Band
Lymph Lymph
Mono Mono
Eos Eos
Baso Baso
Atyp Lym Atyp Lym
Meta Meta
Myelo Myelo
Promyelo Promyelo
Blast Blast
NRBC NRBC

Page 1



Sensitivity (Optional)

Clinical tests are used to identify the presence of disease (sensitivity), as well as the
absence of disease (specificity). This study is performed to determine the sensitivity
and specificity of the XS-Series analyzer using the laboratory’s current numerical
criteria for abnormal samples and the presence or absence of flags generated by the
XS-Series when analyzing normal and abnormal samples. Samples from correlation
studies may be used for this study. Mathematical calculations to determine specificity,
sensitivity, efficiency, false positive rate and false negative rate are applied.

A. Procedure

1. Determine the laboratory’s review criteria to be used for this study. The
review criteria is defined as any numerical criteria that would trigger further
intervention, such as repeating the sample or performing a confirmatory test
such as a manual differential, etc. to verify a result. Please include limits for
Bands, Metas, Myelos, Pros, Blasts, ATLs and NRBCs (i.e. Bands >10% are
judged positive). A Manual Review Criteria Worksheet follows for
documentation.

2. Set the XS-Series Interpretive Program (IP) messages based on the review
criteria prior to analyzing specimens.

3. Criteria for selection and analysis of samples for Sensitivity studies are the
same as the Correlation Studies. Refer to Correlation for data collection
steps. Perform sensitivity calculations on approximately equal numbers of
negative and positive samples. (Example: 100 normal, 100 abnormal)

Note: In addition to retaining printouts from each sample analysis, print the
Q-Flag screen or record the Q-Flag numbers next to each of the flags
generated on the Graphic Printout if adjustment of the IP Suspect flags
is anticipated.

4. Classify each sample result as to “normal” and “abnormal” according to the
laboratory’s criteria using the manual differential and numerical data. Write
this judgment on the XS-Series printout.

5. Review each printout and judge whether an IP message correctly identified
the sample according to Categorizing Negative and Positive Samples
table below. For example, if the laboratory criteria judged the sample
abnormal, and the XS-Series produced a message or flag, the sample is
categorized as a True Positive.

Categorizing Negative and Positive Samples

Category Judgment by Laboratory XS-Series IP messages/Flags

Review Criteria

True Negative (TN) Normal No Flags

True Positive (TP) Abnormal Flag present
False Negative (FN) Abnormal No Flags

False Positive (FP) Normal Flag present

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XS-Series Implementation Manual Section 7: Validation Protocols

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A. Procedure (continued)

6. Tally the total of TN, TP, FN, and FP judgments and calculate the Efficiency,
Sensitivity, Specificity, False Positive and False Negative rate of the XS-Series
using the following formulas.

Efficiency = TP+TN x 100

TN+TP+FN+FP

Sensitivity = TP x 100
TP+FN

Specificity = TN x 100
TN+FP

False Negative Rate = FN x 100
FN+TP

False Positive Rate = FP x 100
FP+TN

7. Definitions of Terms

a. Efficiency (Agreement) The percentage of samples correctly classified by
the XS-Series

b. Sensitivity The ability of the XS-Series to detect abnormalities

c. Specificity The ability of the XS-Series to identify the “normal” samples

d. False Negative Rate The percentage of samples incorrectly classified as
“negative” by the XS-Series

e. False Positive Rate The percentage of samples incorrectly classified as
“positive” by the XS-Series

Note: Sensitivity Studies will not be completed if less than 100 samples have
been processed, and if less than 400 cell manual differentials have
been performed.

References
1. Reference Leukocyte Differential Count (Proportional) and Evaluation of Instrumental

Methods, CLSI/NCCLS Document H20-A March, 1992.
2. Assessment of the Clinical Accuracy of Laboratory Tests Using Receiver Operating

Characteristic (ROC) Plots; Approved Guideline CLSI/NCCLS Document GP10-A, December
1995.

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 18 of 26

Abnormal Manual Review Criteria Worksheet
Manual
Review Using your manual differential and morphology criteria for blood film
Criteria review, define what findings are considered abnormal or “positive”.
For example, a laboratory may define a sample as abnormal for their
population when greater than 10% Atypical Lymphs are present,
where other laboratories may consider > 5% to be abnormal.
Use the table below to quantify the criteria for abnormal samples in
your laboratory.

Differential/Morphology Sample is abnormal when:

Atypical Lymphs
Band/Stab neutrophils
Metamyelocytes
Myelocytes
Promyelocytes
Blasts
NRBC
Abnormal PLT morphology
Abnormal RBC Morphology
Other:
Other:

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This Page Left Blank Intentionally.

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Reference Range (for Customer Reference Only)

Reference ranges describe analyte levels associated with persons who are considered
“healthy”. Multiple factors can influence a reference range, population selection
(geography, and demographics), specimen collection and handling, to name a few.
Assuming the study that resulted in the current reference range was performed
correctly, that the population hasn’t changed and the new instrument is determined to
be comparable (correlation), a study to verify the current reference ranges with the new
analyzer is appropriate. The outcome of this study either verifies the current reference
range or determines whether a new reference range needs to be established.

A. Sample Selection

1. Use your current reference ranges for the CBC and differential parameters.
Establish criteria for “healthy” individuals to select samples. For example:

a. Normal donors may be defined as ones who have no clinical evidence of
a medical disorder known to affect the CBC and differential.

b. Have had no recent episode of bleeding or infection.
c. Have CBC and differential counts within the current reference ranges.
d. Healthy individuals selected should reflect the laboratory’s patient

population, including both male and female of an established age range.
Separate reference ranges based on age may be necessary for pediatric
or geriatric populations.

2. The sample size for this study is dependent upon your laboratory’s policy and
procedure for new analyzer implementation. Select at least 25 to 50 samples.
A larger sampling; however, improves the confidence of the statistics
generated from the study.

B. Analysis Procedure

1. Analyze no more than 25% of the total sample size on one given day.

2. Analyze CBC blood samples on the XS-Series within 4 hours.

3. Analyze the samples on the XS analyzer using the CBC+DIFF discrete order.

C. Statistical Analysis

1. Construct a frequency distribution graph for each of the following parameters,
WBC, RBC, HGB, HCT, MCV, MCH, MCHC, RDW-SD, RDW-CV, PLT, MPV,
the differential percentages and absolute counts.

2. If the distribution is Gaussian, calculate the mean, mode, median, standard
deviation (S.D.) and coefficient of variation (CV%). With Gaussian
distributions, the mean, mode and median should be almost the same

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XS-Series Implementation Manual Section 7: Validation Protocols

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C. Statistical Analysis (continued)

Note: Generally the coefficient of variation is greater than 15-20 % in
non-Gaussian distributions. If the distribution is non-Gaussian, logarithmic
transformation and more sophisticated statistical analysis of the data
would be necessary before calculating the normal range.

3. Calculate the reference range for each parameter using +/- 2 S.D. including
95% of the population.

4. Compare the calculated reference range to the current ranges.

References
1. How to Define and Determine Reference Intervals in the Clinical Laboratory; Approved

Guideline, Second Edition, CLSI/NCCLS C28-A2 Vol. 20, No. 13.

2. Brown, Barbara: Hematology Principles and Procedures, Lippencott, Williams & Wilkins,
Sixth Edition, 1993. p. 32-33.

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 22 of 26

Stability Study (for Customer Reference Only)

Stability studies are performed to determine the readiness of a sample for CBC and
differential analysis. Short term stability is performed with fresh samples drawn and
analyzed at intervals within one (1) hour. Long term stability is conducted under storage
conditions and over a period of time, defined by the laboratory as acceptable for
specimen analysis. Typical long term studies include analysis of room temperature
(18-26 ° C) and refrigerated (4° C) samples at intervals from zero to 48, 56 or 72 hours.

A. Samples for Stability Studies

1. The number of intervals, storage temperatures and the analysis mode chosen
for both short and long term studies determine the amount of blood that must
be drawn from each volunteer.

a. Manual analysis aspirates 20 µL of sample, the auto-mode aspirates 20 µL,
and requires at least 1.0 ml in the tube.

b. Calculate the minimum volume of each sample required for the selected
number of analysis intervals.

c. Double the amount of blood needed for the selected number of intervals
in the long term study if both room temperature and refrigerated samples
will be analyzed.

2. Draw two (2) or more 5 ml EDTA collection tubes from 10 volunteers. Batch
these samples in groups of 3-5 for manageable timing.

3. For long term studies, pool all but one of the collection tubes for each
individual donor.

a. Reserve the single collection tube for the room temperature analysis
intervals (short term and long term, if the volume is sufficient).

b. Label an empty red top tube (no anticoagulant) for each refrigerated
interval.

c. Aliquot the remaining pooled sample into the labeled tubes.

d. Store refrigerated sample aliquots together throughout the study.

e. Record the refrigerator and room temperature.

B. Analysis Intervals

1. Short Term

a. Analyze samples immediately (zero time) then at 5, 15, 30 and 60 minute
intervals.

b. Leave the samples at room temperature throughout the study.

2. Long Term

a. Analyze samples both room temperature and refrigerated samples at zero,
4, 8, 12, 16, 24, 36, 48, 56, 72 hours.

b. Allow refrigerated samples to come to room temperature 15 minutes before
analysis.

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XS-Series Implementation Manual Section 7: Validation Protocols

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Stability Study (continued)

C. Data Analysis
1. Graph results of the following parameters versus time. WBC, RBC, HGB,
HCT, PLT, NEUT%, LYMPH%, MONO%, EO%, BASO%.
2. Graph the short term results versus time in minutes.
3. Graph the long term results versus time in hours for the refrigerated samples
and a separate graph for the room temperature samples.
4. Review the results for when significant loss of stability occurs.

References
1. Hematology Research Center at Loma Linda University School of Medicine.
2. Gould, N. et al., Performance Evaluation of the Sysmex XE-2100 Automated Hematology

Analyzer. Sysmex Journal International: 9: 120-128, 1999.
3. Walters, J., Garrity, P. Performance Evaluation of the Sysmex XE-2100 Hematology

Analyzer, Laboratory Hematology 6:83-92, 2000.

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XS-Series Implementation Manual Section 7: Validation Protocols

Document Number: MKT-30-1009 November 2008 Page 24 of 26

Specimen Mixing Study – XS-1000i with Sampler only
(for Customer Reference Only)

The current CAP Checklist (April 2005) contains the question “Are all blood specimens
collected in anticoagulant for hematology testing mixed thoroughly immediately before
analysis?” A suggested procedure, for demonstrating the adequacy of the Sysmex
XS-Series auto sampler mixing, is to compare the initial RBC values of samples that
were mixed by the auto sampler, to those from the same samples that settled and were
mixed again using the auto-sampler.

Procedure

1. Obtain 10-15 normal and abnormal EDTA anti-coagulated blood samples.

2. Analyze the blood samples on the XS-Series auto sampler three (3) times within
one (1) hour of collection. The auto sampler inverts each sample the (10) times
per analysis.

3. Average the three (3) RBC counts for each of the samples to establish the
baseline mean values.

4. Allow the samples to sit in the auto sampler racks at room temperature for 4 hours.

5. Re-analyze the bloods once on the XS-Series auto sampler. Label the results as
Test 1.

6. Invert each of the samples ten (10) times end-to-end by hand, replace in the
racks and immediately analyze the samples again on the auto sampler. Label the
results Test 2.

7. Compare the baseline RBC values to the RBC values obtained from Test 1 and
Test 2. If results from Test 1 and Test 2 are within 1.5% of the baseline result,
ten (10) inversions are adequate to mix a sample after settling for four hours. If
the variation is greater than 1.5%, allow samples to settle a shorter period of time
and repeat the mixing study.

8. Document the longest period of time samples may settle and still be adequately
mixed by ten (10) inversions.

Reference
1. Gauthier, Steve. Specimen Mixing on the NE-8000, Sysmex Specifics, March 1990. Baxter

Healthcare Corporation.

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This Page Left Blank Intentionally.

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XS-Series Implementation Manual Section 7: Validation Protocols

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Document Number 61-1005
06/2007

Sample Mixing Studies

Sysmex Automated Hematology Analyzers

Introduction

Sysmex automated hematology analyzers are equipped with automated sample-mixing
devices. Each sample is mixed gently, end-to-end, a total of ten times. Studies
performed by Sysmex America, Inc. indicate that the automated mixing time is sufficient
to homogenously disperse the cells in a settled specimen. Excessive mixing of samples,
such as with the use of rotators or rockers, may have adverse effect on the ultimate
results. Therefore, do not pre-mix samples on rotators or rockers before analysis on
Sysmex Hematology analyzers. The procedure used and the resulting data to support
this claim are attached.

Mixing Study Procedure

At fifteen to twenty different sites, 10-15 fresh whole blood samples for CBC analysis
were collected. The samples were analyzed on one of several Sysmex automated
hematology analyzers, using the automated mixing device and the RBC counts were
recorded. The samples were put aside to settle for approximately four hours at room
temperature. The four-hour-old samples were then loaded onto the Sysmex analyzer to
be processed once again. After the allotted time, RBC counts were recorded, the
samples were inverted ten times end-to-end by hand and placed on the Sysmex
analyzer to be analyzed one more time. RBC counts were recorded once again after re-
mixing. The average of all RBC results must fall within the manufacturer’s RBC
Precision specifications CV (1.5%) to be acceptable.

Results

The study included mixing study data from Sysmex automated hematology analyzers
including the Sysmex SF-3000™, Sysmex SE-Series, Sysmex K-4500™, Sysmex
XE-Series, Sysmex HST-Series, Sysmex SE-Alpha™, Sysmex Alpha (R)™, Sysmex
Alpha II™, Sysmex XS-Series, Sysmex XT-Series, Sysmex HST-N Series and Sysmex
XE- Alpha N™ models in 2006/2007. See Table #1 and #2 for results. Each laboratory
site in the study represents 10-15 samples, for a minimum of 200 or a maximum of 300
samples tested. The average of each study is shown in the table with the grand mean
shown as “Mean of All”. RBC counts at all sites were within 1.5% of the manufacturer’s
RBC precision specifications.

Sysmex K-1000 Sysmex SE-Alpha Sysmex XT-2000i
Sysmex KX-21 Sysmex Alpha (R) Sysmex XT-1800i
Sysmex K-21N Sysmex Alpha II Sysmex XE-Alpha
Sysmex K-4500 Sysmex SF-3000 Sysmex XE-Alpha N
Sysmex SE-9000 Sysmex XE-2100 Sysmex HST-Series
Sysmex SE-9500 Sysmex XE-2100L Sysmex HST-N Series
Sysmex SE-9500 (R) Sysmex XS-1000i AL

Page 1 of 2

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1-800-3SYSMEX (800-379-7639)

Document Number 61-1005
06/2007

Conclusion

The specimen mixing on Sysmex automated hematology analyzers is sufficient to
ensure reproducibility of CBC results.

This bulletin was originally released as Roche Bulletin 235 00 0702 on 09/2004.

Page 2 of 2

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1-800-3SYSMEX (800-379-7639)

XS-1000i without Sampler Training Outline

I. XS-1000i System Configuration

• Information Processing Unit (IPU)

• Main Unit (MU)

• Laser Printer

II. XS-1000i Basic Parts

A. Main Unit External Parts
1. Power Switch
2. Open/Close Button
3. Start Switch
4. Ready LED
5. Sample Tube Adaptors
6. Handheld Barcode Reader
7. Locking Screw

B. Main Unit Internal Parts Aspirates sample for testing

1 Piercer Needle Rinses manual aspiration tip (inside and out)
2 Rinse Cup
3 WBC/DIFF Reaction Prepares diluted samples for analyzing WBC
and DIFF. Samples are then sent to optical
Chamber detector block.

4 RBC/PLT Detector Impedance counting enhanced with
hydrodynamic focusing
5 Trap Chamber
6 Air Pump Check daily for fluid accumulation

Replaced every 30000 cycles

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XS-Series Implementation Manual Section 8: Training Tools & Checklist

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XS-1000i without Sampler Training Outline

II. XS-1000i Basic Parts (continued)

C. Graphic Display

1. Review Information

• POSITIVE/NEGATIVE
¾ Positive (backlit red) requires review

Diff – WBC differential count abnormality

Morp – Abnormal Cell Morphology (WBC or RBC)

Count – Abnormal Blood Cell Count

¾ Negative (backlit green) no abnormalities

• ERROR (backlit red) E

¾ Func. : Indicates analysis error not caused by sample
(not including ID read errors)

¾ Result : Indicates error has been caused by the sample, such as
”Sample Not Asp”, Low Blood Volume”, “Low Count Error”.

2. Flags
@ Result falls outside of instrument linearity limits
* Data may be unreliable due to abnormality in sample
+ /- Data outside reference interval

Note: Above indicators are in order of priority.

3. Action Display

• Double-click the “Action” button (in red) on Browser screen to display
the details of the action to be taken.

¾ Count DIFF-CH = To improve the accuracy of the white count,
add DIFF to the discrete testing, and perform the analysis again.

D. Linearity vs. Display Range

Parameter Linearity Display
WBC 0 - 400 x 103/µL 0.00-999.99 x 103/µL
RBC 0 - 8.0 x 106/µL 0.00-99.99 x 106/µL
Hct
Hgb 0 - 60 % 0.0-100.0 %
PLT
0 - 25 g/dL 0-30.0 g/dL
0 - 5000 x 103/µL 0-9999 x 103/µL

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XS-Series Implementation Manual Section 8: Training Tools & Checklist

Document Number: MKT-30-1009 November 2008 Page 2 of 34

XS-1000i without Sampler Training Outline

E. Function Keys
• F1 – Help (also used to silence alarm)
• F2 – Manual
• F4 – Main Menu
• F5 – QC Files
• F6 – Worklist
• F7 – Explorer
• F8 – Browser

III. Reagents

A. Message Displayed “Replace Container -------“
EPK CELLPACK™
FFD STROMATOLYSER-4DL™
FFS STROMATOLYSER-4DS™
SLS SULFOLYSER™

Note: Replace reagents only when Message is displayed. Open dating for all
reagents is 60 days.

B. Replacing a Reagent
1. When XS alarms, press [F1] to silence the alarm. The Help dialog box will
display the reagent that needs to be replaced.
2. Click [Execute] to open Reagent Replacement screen.
3. Verify which reagent to change.
4. Scan the barcode (EAN-128) on the reagent box with the hand-held
barcode reader. The updated information is displayed. If the barcode
cannot be read, the information can be entered manually.
5. Replace the reagent container using clean technique to avoid
contamination.
6. Click [Execute] on the reagent replacement screen to register the
reagent replacement onto the reagent log and start aspiration of the
reagent.

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XS-Series Implementation Manual Section 8: Training Tools & Checklist

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XS-1000i without Sampler Training Outline

IV.Start Up XS-Series

Power Up Sequence:
1 – IPU, 2 – Main Unit, 3 – Printer (if turned off)

IPU: Power on IPU, when the log-on box for the XS Series program
appears, type “XS” for the User Name, and then press [Enter].
There is no password

Main Unit: 1. Do not turn on the Main unit until after the log-on screen on the
IPU displays. Switch is located on the upper right hand side of
the Main Unit.

2. Once the instrument is turned on, it will perform several self
checks (microprocessor, mechanical, temperature and
background).

3. If the background check is successful, the “Analyzer Icon” at the
bottom of the IPU screen and the “Ready LED” on the analyzer
will turn green, and the system is ready for analysis.

If the background exceeds limits, a Background Error message and the Blank
Check dialog box appear. Click [Accept] on the error dialog box to initiate
another Autorinse. Error disappears when background is lower than limits shown
in Acceptable Background Limits table.

Acceptable Background Limits

RBC <0.02 x 106/µL
Hgb
PLT <0.1 x g/dL
WBC-C <10 x 103/µL
WBC-D <0.10 x 103/µL
<0.10 x 103/µL

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V. Quality Control

• QC material should be warmed to room temperature for 15-20 minutes.

• Mix controls by gentle end-to-end inversion until re-suspended.

• Do Not place control material on mechanical mixer or rocker.

• Commercial control files must be established prior to analysis.

Note: Open vial expiration is 14 days for e-CHECK XS (1.5 ml vial) and 7 days
for e-CHECK/e-CHECK(XS) (4.5 ml vial).

Processing QC

A. Manual Mode (4.5 ml or 1.5 ml vials)

1. From the Main Menu, with Ready LED green, click the [Manual] icon or
press [F2].

2. Enter the QC lot number into the “Sample No.” field by one of the
following methods:

¾ using the handheld barcode reader or

¾ manually type in the number using the keyboard or

¾ click the [QC] button in the Manual Sample No. dialog box and
highlight the control level and lot number to be analyzed.

Note: If manually typing the lot number, “QC” must always be upper
case and followed with a dash.

3. Click [OK] or press [Enter] on the keyboard. The QC dialog box opens.

Note: If the lot number is not established, the QC dialog box will fail to
display.

4. Verify appropriate lot and level is selected.

5. Select the appropriate sample tube adaptor and place it in the sample
position area of the XS-1000i with the red mark facing up. Turn the
adaptor clockwise until there is a click (about 45 degrees) to secure it.

6. Place the QC sample into the sample adaptor and press [START] on the
right above the sample position cover.

7. The QC results will display in the dialog box when sample measurement
has completed. Out of range results will display with a red background.
Press [Accept] for results to plot, or [Cancel] to abort.

8. Repeat steps 1-7 for the remaining levels of QC to be analyzed.

One Nelson C. White Parkway, Mundelein, IL 60060 Phone 847-996-4500 · 1-800-3SYSMEX (1-800-379-7639) www.sysmex.com

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V. Quality Control (continued)

Reviewing QC

A. Viewing Radar Charts

1. On IPU, click [QC Files] icon or press [F5].

2. Click the appropriate file number (1-20 or X-barM).

3. Once the file is selected, the Radar Chart will display on the right side of
the screen.

4. The control data is displayed in blue.

5. Date and time of analysis are displayed to the left in the “Analysis Date”
list.

6. Parameters that exceed acceptable limits will have a red “X” displayed on
the Radar chart. The parameter name will be backlit in red and an error
message will be displayed to the left of the file number.

B. Viewing L-J Charts

1. On IPU, click [QC Files] icon or press [F5].

2. Double-click on the appropriate file (1-20 or X-barM). L-J chart will open
when selected.

3. Parameters that exceed acceptable limits will have a red “X” displayed as
the plotted point. The parameter name as well as the result value will be
backlit in red.

4. Scroll down to view remaining parameters using the scroll bar on the right
side of the screen or the down arrow on the keyboard.

One Nelson C. White Parkway, Mundelein, IL 60060 Phone 847-996-4500 · 1-800-3SYSMEX (1-800-379-7639) www.sysmex.com

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VI.Sample Processing

Sample Aspiration Volumes –

♦ Manual Mode = 20 µL minimum volume = 500 µL or 90 µL
♦ Capillary Mode = 67 µL for micro collection

minimum volume = 140 µL

A. Manual Mode

1. When “Ready LED” is green click, [Manual] icon or press [F2].

2. Enter sample ID # using the handheld barcode reader or manually type in
the information using the keyboard.

3. Select Discrete Testing Mode ONLY if Host is not providing that
information.

4. Click [OK] or press [Enter] on the keyboard.

5. Select the appropriate sample tube adaptor and place it in the sample

position area of the XS-1000i with the red mark facing up. Turn the
adaptor clockwise until there is a click (about 45 degrees) to secure it.

6. Place well-mixed sample into the appropriate size adaptor.

Note: If using non-pierceable microtainers, the cap must be removed
prior to processing.

7. Press [START] switch.

B. Capillary Mode

1. Prepare 1:7 dilution (20µL whole blood + 120 µL CELLPACK™).

2. With “Ready LED” green, click the [Manual] icon or press [F2] on the
keyboard.

3. Enter the sample ID number in the “Sample Number” field using the
keyboard or handheld barcode reader.

4. Select [YES] at the Capillary prompt for the mode of analysis.

5. Select discrete test order if this information is not being provided by the host.

6. Click [OK] or press [Enter] on the keyboard.

7. Select the appropriate sample tube adaptor and place it in the sample
position area of the XS-1000i with the red mark facing up. Turn the
adaptor clockwise until there is a click (about 45 degrees) to secure it.

8. Place the well mixed 1:7 dilution into the sample adaptor and press the
[START] switch.

9. The XS will automatically multiply the results by 7.

10. Once testing is completed, reset the capillary mode radio button back to [NO].

One Nelson C. White Parkway, Mundelein, IL 60060 Phone 847-996-4500 · 1-800-3SYSMEX (1-800-379-7639) www.sysmex.com

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VII. Maintenance

A. Daily Shutdown – Performed Daily to clean detector and dilution lines

1. Click [Shutdown] icon on the Menu screen.

2. Click [Execute] to begin the process. The procedure will take
approximately 2 minutes to complete.

3. Once completed, the Shutdown dialog box will be replaced by the
Power off dialog box.

4. Click [Restart] to initiate an Autorinse and continue sample analysis
or perform a complete power off if further testing is not going to be
done. (This is recommended at least once a week.)

5. If performing a complete “Power Off”, power off the MU, then on the
IPU click [File], [Exit].

6. Click [YES] to exit the XS program.

7. Click [Start] at the bottom of the Windows desktop.

8. Click [Shutdown] in the [Start] Menu.

9. A window appears with the option to choose shutdown. Press [Enter].
This allows the system to write unsaved data to the hard drive and
power itself off.

B. Monthly Rinse – Every 1200 Cycles or Monthly

1. Click [Controller] icon on the Main Menu.

2. Click [Maintenance] icon.

3. Click [Monthly Rinse] icon a dialog box will appear.

4. Place a tube containing a minimum of 3 mls of CELLCLEAN™
(5% Sodium Hypochlorite Solution) into the sample position with the
appropriate sample tube adaptor.

5. Press [START] to initiate cleaning.

6. Once completed, the Monthly Rinse dialog box is replaced by the
Power off dialog box.

7. Click [Restart] to initiate an Autorinse and continue sample analysis
or perform a complete power off if further testing is not going to be
done.

C. Every 30,000 Cycles

1. Change Piercer

2. Replace Air Pump

One Nelson C. White Parkway, Mundelein, IL 60060 Phone 847-996-4500 · 1-800-3SYSMEX (1-800-379-7639) www.sysmex.com

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VII. Maintenance (continued)

D. Setting Cycle Counters
1. Click [Controller] icon on the Main Menu.
2. Click [Maintenance] icon on the Controller Menu.
3. Click [Counter] icon on the Maintenance Menu.
4. Click [Reset] on the Piercer or Air Pump box, and then click [OK] to
reset the counters to zero.
5. Click [Menu] on the Toolbar or [F4] to return to the Main Menu Screen.

E. As Needed Maintenance
1. Clog Removal
2. Clean RBC Aperture
3. Remove Air Bubbles
4. Rinse Flow Cell

VIII. Principles and Technology

RBC Hydrodynamic focusing + impedance
Hgb Sodium Lauryl Sulfate

Hct Cumulative Pulse Height measurement

MCV Calculated from Hct and RBC

PLT-I Hydrodynamic focusing + impedance
Diff Channel
WBC-C Side Scatter (internal complexity) &
WBC-D Fluorescence (RNA/DNA)

Forward Scatter (cell size)

Sideways fluorescent light

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