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LH500, Beckman/Coulter; Operate

5C Control

Change Report Type

Controls Manual Aspirate

Expected Results

Instrument Flags

Latron Primer/Control

Limitations

Materials

New Latron Control

New Lot 5C

Patients Auto Aspirate

Patients Manual Aspirate

Quality Control

Reagent Update

Reagents

Result Derivation

Samples Pre-Dilute

Specimen

Transmit Results

I. Principle:

The Beckman/Coulter LH500 is an in vitro analytic instrument that provides complete blood cell (CBC) counts and white blood cell (WBC) differential counts on whole blood samples.

System Methodologies:

  1. WBC: Impedance counting. Pulses measured in the WBC aperture representing cells as 35fl or greater are classified as white blood cells.
  2. RBC: Impedance counting. Pulses measured in the RBC aperture representing cells as 36fl or greater are classified as red blood cells.
  3. Platelet Count: Impedance counting. Pulses measured in the RBC aperture representing cells from 2-20 fl are classified as platelets.
  4. HGB: The transmittance of light (525nm wavelength) through the lysed WBC solution in the hemoglobin cuvette is compared to the transmittance of the same light through a reagent blank. The system converts this ratio to a HGB value in g/dl using a calibration factor.
  5. MCV: The average volume of individual erythrocytes derived from the RBC histogram.
  6. RDW: The size distribution spread of the erythrocyte population derived from the RBC histogram.
  7. MPV: The average volume of individual platelets derived from the PLT histogram.
  8. HCT, MCH and MCHC: Calculated parameters.
    HCT: The relative volume of packed erythrocytes to whole blood. HCT = RBC x MCV ÷10.
    MCH: The weight of HGB in the average erythrocyte. MCH = HGB ÷ RBC x 10
    MCHC: The average weight of HGB in a measured dilution. MCHC = HGB ÷ HCT x 100
  9. WBC Differential: The instrument makes 3 measurements (volume, conductivity and scatter) as each cell passes through the flow cell. The low frequency impedance measurement defines cell volume. The high frequency conductivity measurement indicates the internal conductivity. The light scatter measurement indicates the structure and shape. The AccuGate algorithm is applied to determine different cell populations.

II. Materials:

Racks

III. Reagents:

  1. Coulter LH Series Diluent (Beckman/Coulter #8547194)
    1. Store at 15 - 30°C. Stable unopened until the expiration date printed on the label.
    2. Stable for 60 days after opening.
    3. Mixing instructions:
      1. Roll the cube 20 times.
      2. Let it set 24 hours to de-bubble.
  2. Coulter LH Series PAK Reagent Kit (Beckman/Coulter #8547195)
    contains Erythrolyse II and StabiLyse
    1. Store at 15 - 30°C. Stable unopened until the expiration date printed on the label.
    2. Stable for 60 days after opening.
  3. Coulter Lyse S III diff Lytic Reagent (Beckman/Coulter #8546983)
    1. Store at 15 - 30°C. Stable unopened until the expiration date printed on the label.
    2. Stable for 60 days after opening.
  4. Coulter Clenz Cleaning Agent (Beckman/Coulter #8546931)
    1. Store at 15 - 30°C. Stable unopened until the expiration date printed on the label.
    2. Stable for 90 days after opening.
  5. Latron Primer (Beckman/Coulter #7546915)
    1. Store at 15 - 30°C. Stable unopened until the expiration date printed on the label.
    2. Stable for 30 days after opening.
  6. Latron Control (Beckman/Coulter #7546914)
    1. Store at 15 - 30°C. Stable unopened until the expiration date printed on the label.
    2. Stable for 30 days after opening.
  7. Coulter 5C Cell Control (Beckman/Coulter #7547116)
    1. Store at 2 - 8°C. Stable unopened until the expiration date printed on the label.
    2. Stable for 13 days after opening.
    3. Allow vials to equilibrate at room temperature for 15 minutes.
    4. Mix each vial using the 8x8x8 method.
      1. Hold the vials with the stoppers up and roll in hands 8 times.
      2. Hold the vials with the stoppers down and roll in hands 8 times.
      3. Gently invert the vials end to end 8 times.
    5. Repeat steps D.a-c.
    6. Visually check each vial after mixing to ensure cells are suspended. If cells are not suspended, then gently invert the controls 8 more times.

IV. Specimen:

CBC/CBC Diff: Samples should be mixed for 5 minutes before running. Refer to "Specimen Integrity, Hematology" in the Quality Assurance Manual.
Automatic Aspiration Mode

Manual Aspiration Mode

Prediluted Mode

V. Reagent Update:

  1. Record the new reagent information:
    1. Click on the System Setup icon. (Yellow diamond, located in the lower left corner of the screen.)
    2. Select QA.
    3. Select the Reagent Tab.
    4. Click on Reagent Setup.
    5. Use the handheld barcode reader to scan the 2 barcodes on the new reagent container. (The barcodes may be scanned in any order.)
    6. Verify that all the fields of the “Reagent Information Preview” box have populated correctly.
    7. Click the Save button (green check mark) to complete the importing of the new reagent data.
  2. Turn OFF the Compressor (if it is ON).
    1. Select the Instrument Computer.
    2. Select Diagnostics, then Operator Options and then Fluidics Tests.
    3. Highlight “Compressor ON/OFF”, and press Enter.
    4. Press the Spacebar to select OFF.
    5. Press Enter.
  3. Open the new reagent container.
  4. Unscrew the pickup tube assembly and slide the collar off the old reagent container.
  5. Lift the assembly straight up and out. Be careful not to touch part of the assembly or let it touch any lab surfaces.
  6. Transfer the pickup tube assembly to the new reagent container and secure it.
  7. Prime the reagent line:
    1. Select the instrument computer.
    2. Select Diluter Functions and then Prime Reagents.
    3. Select the reagent that was replaced and press Enter.
      Note: While new reagent is priming, an alarm may sound. Return to the workstation computer and select Workstation OK on the warning screen to silence the alarm.
  8. When priming is complete, select F9 to exit.
  9. Return to the Workstation computer.

VI. Running Latron Primer and Control:

  1. Select the Instrument Process type on the Command Center as CONTROL.
  2. Select the Run Type as Ltx Prime. The instrument will automatically change the Aspiration Mode to Manual.
  3. Click on the START button.
  4. Mix the Primer bottle by gentle inversion. Be careful not to introduce any bubbles into the bottle.
  5. Aspirate the Latron Primer in the manual aspiration mode.
  6. Review the Latron Primer results on the QA screen.
    1. Select “QA” on the Command Center to display the Quality Assurance application.
    2. Select “QC” to display the control windows.
    3. Select the Latron Control by double-clicking on Latron on the left side of the screen.
    4. Single-click on the lot of Latron and the control results table, statistics and graphs appear on the window. Use the scroll bars on the window to view other parameter results and graphs.
  7. The results for the Primer must be ≤ 500. The control trial will be highlighted in red if it is unacceptable. If the Latron Primer results are unacceptable, repeat the Primer once. If repeat is still unacceptable the flowcell may need to be purged, or the Primer may have excessive bubbles.
  8. Click on the STOP button.
  9. Select the Run Type as Latex.
  10. Click on the START button.
  11. Mix the Latron Control bottle by gentle inversion. Be careful not to introduce any bubbles into the bottle.
  12. Aspirate the Latron Control in the manual aspiration mode.
  13. Review the Latron Control results on the QC screen. The control trial will be highlighted in red if results are unacceptable. If the Latron Control results are unacceptable, repeat the control once. If the repeat is unacceptable, Purge the Flowcell and then repeat the Control. Refer to “PURGE the Flowcell” in HELP.
  14. Click on the STOP button.
  15. Change the Instrument Process Type to AUTOANALYSIS.
  16. Change the Run Type to “CD”.
  17. Change the Aspiration Mode to AUTO.

VII. Running 5C Cell Controls (Automatic Aspiration Mode):

  1. Allow the controls to reach room temperature and mix according to handling instructions in Reagents section. The controls should reach room temperature in approximately 15 minutes.
  2. Place control vials in a rack with the barcode labels facing up.
  3. Place the rack in the autosampler. If the instrument is in the Ready Mode, it will automatically start cycling. If the START button is green, click on it to start the system.
  4. Review the control results in the QA file.
    1. Select “QA” on the Command Center to display the Quality Assurance application.
    2. Select “QC” to display the control files.
    3. Double-click on 5C Series folder on the left side of the screen and single-click to select the specific control for the results to be reviewed. The control results table, statistics and graphs appear on the window. Use the scroll bars on the window to view other parameter results and graphs.
  5. Unacceptable Control Trials are highlighted in red. Individual constituents are flagged with “L” or “H” to indicate the constituent as unacceptable low or high respectively.
  6. If controls are unacceptable, repeat them once. If controls are still unacceptable, either the control vial may have low volume and a new vial should be opened or maintenance is needed. Check to see if there are any flags associated with the control results. This will help to determine what maintenance may be appropriate to perform first. Refer to section XX.8.

VIII. Running Patient Samples (Automatic Aspiration Mode):

  1. Load samples into the rack with the barcode label facing up.
  2. Place the rack securely into the loading bay on the right side of the instrument. If the instrument is in the Ready mode it will automatically start cycling. If the Start button is green, press the START button to start the instrument.
  3. Review patient results. The instrument has been programmed with “Review by Exception”.
    1. Samples that pass through these filters will be “Auto-validated” and sent to CLICS Instrument Review Release.
    2. Samples that did not pass through the review criteria will be in the “Review List” folder.
  4. To review patient results, select the “Patient Result” icon on the Command Center.
  5. Click on “Patient File Folder” icon located at the top of the screen. The patient results are placed in folders based on status and Run Type.
  6. Select the “Review List” folder. The patient results that did not pass through the review criteria will be in this folder. Results are listed with the most recent at the top.
  7. Highlight the patient results to be reviewed and then select the “Results and Graphics” icon on the right side of the screen. Patient results will be displayed. The CBC Data including histograms, and the Diff Data including cytograms, are available by selecting the appropriate tabs on the patient results screen.
  8. Refer to “Peripheral Blood Smear (PBS) Review, Technologist/Pathologist” for appropriate review steps.
  9. If the barcode label was not read, the sample ID will be displayed as “------“. To enter the Sample ID, select the “Edit” button on the right of the screen. Enter the correct Sample ID and press the Tab key. The edited field will be flagged with an “E”. Click on the “Save Edit” icon on the right side of the screen.
  10. Editing of patient results should be done on the CLICS Instrument Review Release screen.
  11. To print results, select the “Printer” icon on the top left side of the screen.
  12. To release results to CLICS, select the “Validate” button on the top of the screen.
  13. Close out the patient result screen to return to the “Review List” folder.

IX. Running Patient Samples (Manual Aspiration Mode):

  1. Press the STOP button.
  2. Change the Aspiration Mode to Manual.
  3. Press the START button. When the instrument is ready for manual aspiration the cursor will be in the barcode field on the Workstation Command Center.
  4. If the sample has a barcode label:
    Scan the sample barcode using the handheld barcode reader.
  5. If you are entering the Sample ID manually:
    1. Type in the Sample ID to be used.
    2. Press the Tab key.
  6. Remove the stopper from the specimen tube.
  7. Immerse the aspirator tip in the tube. Press the manual aspiration activator.
  8. When you hear a beep, remove the tube from the aspirator tip. The probe cleaner automatically cleans the aspiration probe.
  9. After the instrument cycles the sample, review the sample results on the Workstation. Refer to steps listed in the previous section for sample results review.
  10. Press the STOP button.
  11. Change the Aspiration Mode to Auto.

X. Running Control Samples (Manual Aspiration Mode):

  1. Select the Instrument Process type on the Command Center as CONTROL.
  2. Select RUN type as CD.
  3. Change the Aspiration Mode to Manual.
  4. Insure the cursor is in the barcode field and type in the Control Lot # (not the barcode number) and press the Tab key.
  5. Process the Control in the Manual Mode. Refer to Running Patient Samples/Manual Aspiration Mode.

XI. Running Samples/Prediluted Mode:

  1. Press the STOP button.
  2. Change the Aspiration Mode to Manual.
  3. Select the Prediluted Mode.
  4. A window will be displayed asking for the dilution factor. If a citrate tube is being sampled due to platelet clumps the dilution factor to enter is 1.1. If a 1:2 dilution is being run because a sample has results above the linear range, the dilution factor to enter is 2.0. The instrument will automatically correct the results for the dilution factor entered.
  5. Press the Start button.
  6. If the sample has a barcode label:
    Insure the cursor is in the barcode field at the Workstation Command Center; scan the sample barcode using the handheld barcode reader.
  7. When entering the Sample ID manually, with the cursor in the barcode field at the Workstation Command Center, type in the Sample ID to be used. Press the Tab key.
  8. Remove the stopper from the specimen tube.
  9. Immerse the aspirator tip in the tube. Press the manual aspiration activator.
  10. When the audible beep sounds, remove the tube from the aspirator tip. The probe cleaner automatically cleans the aspiration probe.
  11. After the instrument cycles the sample, review the sample results on the Workstation. Refer to steps listed above for sample results review.
  12. Press the STOP button.
  13. Return the Aspiration Mode to Auto.

XII. Importing New Control Lot Number for Beckman/Coulter 5C Control:

  1. Select the Setup Icon on the Command Center.
  2. Select the QA icon.
  3. Select the Control Tab.
  4. Select the “Set up New Control Folder” icon.
  5. Select BCI- 5C-All Levels.
  6. Select the “Set Up New Lot” icon.
  7. Follow the directions on the pop-up window. Check Auto Stop and Auto Transmit in step 2.
  8. Select OK.
  9. Select the Close icon.
  10. Select OK to close the System Setup/QA window.
  11. When switching to a new lot number of the same control, a 3 day overlap is performed to determine the new target means to be used. The laboratory staff review the data using the following protocol.
    1. A 3 day overlap must be performed when switching to a new lot of control.
      1. The new lot number of controls will be imported according to the above instructions.
      2. A 3 day overlap is performed, running all 3 levels of the new control lot every 8 hours. The new lot number of control and the current lot number of control are run together. The data for each control will be stored in their respective files per control code.
      3. Following the 3 day overlap, the results obtained from the new lot number control are printed and reviewed. Refer to step 12 for printing overlap data.
      4. The printed control data will include calculated mean, SD and CV values for all constituents. A Mean Plot chart will also show where each constituent mean fits within the 2SD limits. If the mean values fall within the expected 2SD ranges, as noted on the Mean Plot chart, then the control target values for each parameter will be edited to the mean value obtained from the control overlap.
      5. If any of the control values fall outside of the expected 2SD limits, the site supervisor must be notified before proceeding.
    2. To apply Lab Limits to the control file and calculate the mean from the overlap data:
      1. Select the specific control for which you want to adjust limits. The control results table, statistics and graphs appear in the window.
      2. Select “Mean=>Lab Target” to replace the assigned values on this window with the values that currently appear as the mean values. The Workstation also replaces the expected ranges. The Workstation now displays the Lab Target and Lab Limit values.
  12. Printing Controls Prior To Deletion:
    Following a successful control overlap, print a hard copy of the old lot# of control prior to deleting from the control dictionary.
    1. Select the QA icon on the Command Center.
    2. Select the QC icon.
    3. Highlight the control lot number to be printed and press the Printer Icon.
    4. Select “Selected Lot” in the List Runs section. Select “Full Graph” in the Levey-Jennings Graph section.
  13. Deleting A Control From The Control Dictionary:
    1. To delete the old control lot numbers, select the QA icon on the Command Center.
    2. Select the QC icon.
    3. Highlight the control lot number to be deleted and click on the Delete icon on the right side of the screen.
    4. Select “All data including set-up tables” and then select “OK”.
    5. A warning message will display: “Are you sure that you have made the right choice?” Click OK if this is the file to be deleted.

XIII. Importing New Control Lot Number for Latron Control

Note: Latron Primer does not require importing new ranges for new lot number. Acceptable range is always <500.

  1. Select the Setup Icon on the Command Center.
  2. Select the QA icon.
  3. Select the Control Tab.
  4. Select the “Set up New Control Folder” icon.
  5. Select BCI- Latron.
  6. Select the “Set Up New Lot” icon.
  7. Enter the information listed on the package insert, starting with the lot number. Use the tab key to advance to the next field.
  8. Select OK.
  9. Select the Close icon.
  10. Select OK to close the System Setup/QA window.
  11. After scanning the assay sheets, ensure that the correct features are still selected or enabled.
    1. Double-click on the yellow diamond icon.
    2. The System Setup window is displayed; double-click on the QA-Quality Assurance icon.
    3. The System Setup (Quality Assurance) window is displayed; click on the Control tab.
    4. All controls are displayed: At the middle of the screen, review “Auto Xmit” and “Auto Stop” columns.
      LH500: There should be an “x” (enabled) in the Normal, Abnormal I and Abnormal II “Auto Stop” and “Auto Xmit” boxes and “x” in both the CControl and MControl “Auto Xmit” boxes. All other boxes should be blank/grey.
    5. If “x”s are not in the above configuration and need to be edited, click on the left side of the line to be edited (should backlight in black) and then click on the “pencil” icon.
    6. The Control Folder Setup window is displayed.
    7. At the top center of the window is an Automatic Data Handler box containing “Auto Transmit” and “Auto Stop” boxes.
    8. Click a check mark into the desired box (or de-select).
    9. Click on the green check mark to close out and Save the selection.
      Note: This must be done every time a new lot number of QC data is imported via the 2D BCR
      .
  12. Printing Controls Prior To Deletion:
    Print a hard copy of the old lot# of control prior to deleting from the control dictionary.
    1. Select the QA icon on the Command Center.
    2. Select the QC icon.
    3. Highlight the control lot number to be printed and press the Printer Icon.
    4. Select “Selected Lot” in the List Runs section. Select “Thumbnail” in the Levey-Jennings Graph section.
  13. Deleting A Control From The Control Dictionary:
    1. To delete the old control lot numbers, select the QA icon on the Command Center.
    2. Select the QC icon.
    3. Highlight the control lot number to be deleted and click on the Delete icon.
    4. Select “All data including set-up table” and then select “OK”.
    5. A warning message will display: “Are you sure that you have made the right choice?” Click OK if this is the file to be deleted.

XIV. Changing Report Type:

In the event that CLICS or the Hospital/Clinic computer system is down, the report type can be changed so the Suspect and Definitive Messages do not print.

  1. Select the Setup icon on the Command Center.
  2. Select the General Settings icon.
  3. Select Reporting Options.
  4. Select Chartable Report as the Printout Option.
  5. Deselect Laboratory Report.
  6. Click on OK.

XV. Transmit Results to LIS:

  1. Select the Patient Results icon.
  2. Select the appropriate patient results folder (Database/To Do List).
  3. Select the patient results to be transmitted.
  4. Click on the “Host Transmission” (Transmit Results) icon on the top left corner of the patient results screen.

XVI. Limitations:

  1. White Blood Cells (WBC): Certain unusual RBC abnormalities that resist lysing, nucleated RBC (NRBC), fragmented WBC, agglutinated WBC, any unlysed particles greater than 35fl, very large or aggregated platelets, specimens containing fibrin, cell fragments, or other debris such as in pediatric and oncology specimens.
  2. Red Blood Cells (RBC): Very high WBC count, high concentration of very large platelets, agglutinated RBC, RBC smaller than 36fl, specimens containing fibrin, cell fragments, or other debris such as in pediatric and oncology specimens.
  3. Hemoglobin (HGB): Very high WBC count, severe lipemia, heparin, certain unusual RBC abnormalities that resist lysing, or anything that increases the turbidity of the sample such as elevated levels of triglycerides.
  4. Mean Corpuscular Volume (MCV): Very high WBC count, high concentration of very large platelets, agglutinated RBC, RBC fragments that fall below the 36fl threshold, or rigid RBC.
  5. RBC Distribution Width (RDW): Very high WBC count, high concentration of very large or clumped platelets, agglutinated RBC, RBC below the 36fl threshold, two distinct populations of RBC, or rigid RBC.
  6. Platelet (PLT): Very small red blood cells near the upper threshold, cell fragments, Giant or clumped platelets, platelet fragments, or cellular debris near the lower platelet threshold, electronic noise.
  7. Mean Platelet Volume (MPV): Known factors that interfere with the platelet count and shape of the histogram or known effects of EDTA.
  8. Hematocrit (HCT): Known factors that interfere with the parameters used for computation: RBC and MCV.
  9. Mean Corpuscular Hemoglobin (MCH): Known factors that interfere with the parameters used for computation: HGB and RBC.
  10. Mean Corpuscular Hemoglobin Concentration (MCHC): Known factors that interfere with the parameters used for computation: HGB, RBC and MCV
  11. Diff Parameters: Known factors that affect the WBC count as listed above or high triglycerides that affect lysing. Hypogranular or agranular granulocytes lyse resistant red cells, very small or multi-population lymphocytes, precipitated elevated proteins.
  12. Samples with cold agglutinins may falsely lower the red blood cell count. Refer to “Cold Agglutinin Artifact in Hematology Analysis (CBC), Prevention of”.
  13. Very high glucose levels (≥ 800 mg/dl) may falsely increase MCV. This results in falsely high hematocrit, and falsely low MCHC.ref 6

XVII. Instrument Flags:

  1. “V” indicates one of the three counts voted out.
    Action:
    The count was calculated using 2 counts. Report result. If this becomes a chronic problem: Clean the aperture using “Multiple Aperature Zap” procedure under Diagnostics/Operator/Fluidics. Refer to “LH500 Help” for procedure.
  2. ”-----“ Indicates Total Voteout (LH500)
    Action:
    Check the sample for clots. If clots are seen, collect a new sample. If no clots are seen, perform the aperture cleaning procedure and then rerun the sample.
  3. “:::::” The instrument detected a clog in the flow cell, PC1, PC2 or FC will be displayed on the y-axis of the dataplot. This could also be due to resistant RBC. If the latter is the case, the flag will be limited to one patient and a manual differential will be required.
    Action:
    If the flag is limited to one patient, check the patient sample for clots. If clots are found, a new sample must be collected. If flag occurs on multiple samples, there is a clog in the flowcell. In either case, clean the flowcell.
    1. “Purge Flowcell” procedure. Refer to “LH500 Help” for this procedure.
  4. “+++++” Result exceeds the instrument’s operating range. Refer to “section XVIII. Results Derivation”.
  5. “.....” Incomplete computation. This could occur on calculated parameters because of a voteout (-----) or over range results(+++++) for the parameters used in the calculation. Refer to “section XVIII. Results Derivation”.
  6. “P” indicates partial aspiration. This occurs in the “Auto” mode when the blood detectors sense a possible clot, or a HGB of approximately ≤ 5 gm/dl.
    Note: If chronic “P” flags occur, contact an Instrument Specialist. The manual mode does not detect partial aspiration. Do not run samples in manual mode in an attempt to clear up the flag. Doing so may mask a problem.
    1. Check the sample for clots.
      1. If no clots are seen, rerun the sample. Refer to “Specimen Integrity, Hematology” protocol.
      2. If clots are seen, collect a new sample.
    2. If HGB is in the 5-6 gm/dl range, the blood detectors may be triggering a “P” flag because of the low HGB.
      1. Release instrument results to CLICS.
      2. Delta Check GO () – release the results.
      3. No Delta Check or Delta STOP (): check the sample for clots.
        1. If no clots are seen, release the results. Refer to “Specimen Integrity, Hematology” protocol.
        2. If clots are seen, collect a new sample.
  7. “R” indicates review. This flag is not used as a criterion for review in this procedure. It may display in conjunction with other flags that are defined.

XVIII. Results Derivation:

  1. Analytical Range: (For trouble shooting purposes only.)
    1. WBC (103/µl): 0.00 - 200.00
    2. RBC (106/µl): 0.00 - 7.00
    3. HGB (g/dl): 0.0 - 25.0
    4. PLT (103/µl): 0 - 2000
  2. Reportable Ranges:
    Note:
    Results above the upper reportable limit must be diluted with LH Diluent and rerun in the Prediluted Mode. The dilution factor must be entered so the instrument can correct the results for the dilution. If the WBC must be diluted, refer to “Hemoglobin Correction for High WBC” procedure” for corrections needed for HGB, HCT, MCH and MCHC.
    1. WBC (103/µl): 0.70 - 223.00 (reported as thou/µl)
    2. RBC (106/µl): 0.30 - 7.30 (reported as mil/µl)
    3. HGB (g/dl): 0.9 – 24.4
    4. PLT (103/µl): 9 - 3151 (reported as thou/µl)
  3. If a glucose ≥ 800 mg/dl occurs on a patient, enter a Canned Comment “MCV may be falsely elevated due to very high glucose, resulting falsely elevated hematocrit, and falsely low MCHC.”ref 5
  4. In the event that a critical value is encountered on a test that is not ordered, the following steps are performed: (Example: A critical WBC is obtained on a sample that has only a HGB/HCT ordered.)
    1. Verify the critical value according to “Critical Value Chart & Protocol”.
    2. Call the nursing unit or doctor’s office and inform them that a critical value (high or low) has been obtained on the patient sample on a test that was not ordered. Report the result following the “Critical Value Chart & Protocol”.
    3. Request that an order be placed for the test with the critical value so that the critical value result can be officially entered in CLICS. Use “Add-On Order” procedure as needed to ensure the new order has the same accession number.
    4. Retransmit results from LH500 to CLICS. Refer to section XV. Transmit Results to LIS.
    5. Refer to “Result Review/Release, Interfaced Instrument” to release results.
    6. Refer to “Phoned/Faxed Result Documentation, CLICS” to document critical value notification.

XIX. Expected Result(s) and/or Critical Values:

WBC: ref 3

sex

age range

expected range

critical low

critical high

M & F

0-1 yr

6.0-17.5 x thou/µl

1.0

50.0

M & F

1-2 yr

6.0-17.0 x thou/µl

1.0

50.0

M & F

2-4 yr

5.5-15.5 x thou/µl

1.0

50.0

M & F

4-6 yr

5.0-14.5 x thou/µl

1.0

50.0

M & F

6-8 yr

4.5-13.5 x thou/µl

1.0

50.0

M & F

8-10 yr

4.5-13.5 x thou/µl

1.0

50.0

M & F

10-16 yr

4.5-13.0 x thou/µl

1.0

50.0

M & F

≥ 16 yr

4.5-11.0 x thou/µl

1.0

50.0

RBC: ref 3

sex

age range

expected range

critical low

critical high

M & F

0-15 days

3.9-5.9 x mil/µl

   

M & F

15 days-1mo

3.3-5.3 x mil/µl

   

M & F

1-4 month

3.5-5.1 x mil/µl

   

M & F

4-6 month

3.9-5.5 x mil/µl

   

M & F

6-9 month

4.0-5.3 x mil/µl

   

M & F

9 month-1yr

4.1-5.3 x mil/µl

   

M & F

1-3 yr

3.8-4.8 x mil/µl

   

M & F

3-6 yr

3.7-4.9 x mil/µl

   

M & F

6-9 yr

3.8-4.9 x mil/µl

   

M & F

9-12 yr

3.9-5.1 x mil/µl

   

M

12-15 yr

4.1-5.2 x mil/µl

   

F

12-15 yr

3.8-5.0 x mil/µl

   

M

15-18 yr

4.2-5.6 x mil/µl

   

F

15-18 yr

3.9-5.1 x mil/µl

   

M

18-45 yr

4.3-5.7 x mil/µl

   

F

18-45 yr

3.8-5.1 x mil/µl

   

M

45-65 yr

4.2-5.6 x mil/µl

   

F

45-65 yr

3.8-5.3 x mil/µl

   

M

≥ 65 yr

3.8-5.8 x mil/µl

   

F

≥ 65 yr

3.8-5.2 x mil/µl

   

HGB: ref 3

sex

age range

expected range

critical low

critical high

M & F

0-15 days

13.4-19.8 g/dl

12.5

22.0

M & F

15 days-1month

10.7-17.1 g/dl

8.0

17.0

M & F

1-2 month

9.4-13.0 g/dl

8.0

17.0

M & F

2-4 month

10.3-14.1 g/dl

8.0

17.0

M & F

4-6 month

11.1-14.1 g/dl

8.0

17.0

M & F

6month-5 yr

11.0-14.0 g/dl

8.0

17.0

M & F

5-9 yr

11.5-14.5 g/dl

8.0

17.0

M & F

9-12 yr

12.0-15.0 g/dl

8.0

17.0

M

12-15 yr

12.0-16.0 g/dl

7.0

20.0

F

12-15 yr

11.5-15.0 g/dl

7.0

20.0

M

15-18 yr

11.7-16.6

7.0

20.0

F

15-18 yr

11.7-15.3

7.0

20.0

M

18-45 yr

13.2-17.3

7.0

20.0

F

18-45 yr

11.7-15.5

7.0

20.0

M

45-65 yr

13.1-17.2

7.0

20.0

F

45-65 yr

11.7-16.0

7.0

20.0

M

≥ 65 yr

12.6-17.4

7.0

20.0

F

≥ 65 yr

11.7-16.1

7.0

20.0

HCT: ref 3

sex

age range

expected range

critical low

critical high

M & F

0-15 days

41-65 %

   

M & F

15 days-1month

33-55%

   

M & F

1-2 month

28-42%

   

M & F

2-4 month

32-44%

   

M & F

4-6 month

31-41%

   

M & F

6-9 month

32-40%

   

M & F

9 month-1 year

33-41%

   

M & F

1-3 year

32-40%

   

M & F

3-6 year

32-42%

   

M & F

6-9 year

33-41%

   

M & F

9-12 year

34-43%

   

M

12-15 year

35-45%

   

F

12-15 year

34-44%

   

M

15-18 year

37-48%

   

F

15-18 year

34-44%

   

M

18-45 year

39-49%

   

F

18-45 year

35-45%

   

M

45-65 year

39-50%

   

F

45-65 year

35-47%

   

M

≥ 65 year

37-51%

   

F

≥ 65 year

35-47%

   

Absolute Reticulocyte Count: ref 3

sex

age range

expected range

critical low

critical high

M & F

0-14 days

239-404 x thou/µl

   

M & F

14 days-12 yrs

46-154 x thou/µl

   

M & F

≥ 12 yrs

59-146 x thou/µl

   

The rest:

Analyte

Expected Range

Critical Low

Critical High

MCV: ref 3

80-100 fl

   

MCH: ref 3

26-34 pg

   

MCHC: ref 3

32-37 g/dl

   

RDW: ref 1

12.1 - 15.2%

   

Platelet Count: ref 4

150-400 x thou/µl

<30

>1,000

Platelet oncology:

 

<10

 

MPV ref 1

7.5 - 11.2 fl

   

Neutrophils: ref 1

42-74%

   

Lymphocytes: ref 1

18-44%

   

Monocytes: ref 1

5-13%

   

Eosinophils: ref 1

0-8%

   

Basophils:ref 1

0-1%

   

Absolute Neutrophils

 

<0.5

 

XX. Quality Control:

  1. Routine Controls:
    1. Beckman/Coulter 5C™ Hematology Controls are assayed every 8 hours of use alternating controls. Two levels of control are assayed following morning start up. One level is then assayed on other shifts. Two levels of controls are also run after replacing reagents. All commercial controls are assayed in the Auto Mode.
    2. In lieu of using commercial control material for the Open Vial Sample Mode, patient whole blood is used as control material. The same patient whole blood sample must be run in both the Auto Mode and the Open Vial Mode every 8 hours of use. An EDTA whole blood patient sample, with results in the normal range, is obtained from available samples.
      1. Run an EDTA sample in the Auto Mode using barcode ID MCONTROL; results will automatically print.
      2. Run the same sample immediately in the Open Vial Sample mode using barcode ID MCONTROL. (It must be the next sample run.) The results will automatically print.
      3. Compare the results of both trials. The results from the Auto and Open Vial modes must match within stated limits. Attach the Auto and Open Vial reports together and sign them indicating results were reviewed and were acceptable.
      4. If results do not match within the stated limits, rerun the sample in both modes. If results are still not within the stated limits contact the Site Supervisor.
      5. Forward the reports to the Instrument Specialist Office at the end of the day.
    3. City Control samples are run once per day. Results are entered into the “City Control Data Acquisition Logs”. Refer to the “Hematology City Control: Preparation, Application and Management” protocol.
    4. The Latron Primer and Control are run once a day, typically following morning Startup.
  2. Criteria for accepting or rejecting a patient run is based on the “Quality Control; General” protocol in the Quality Assurance Manual.
  3. Calibrator: Beckman/Coulter S-CAL Calibrator.
  4. Calibration Frequency: Calibration is performed every 6 months. Calibration may also be performed after major maintenance and/or replacement of critical parts that may influence test performance.
  5. Calibration Verification: Calibration verification is performed every 6 months and whenever there is major preventative maintenance or replacement of critical parts that may influence test performance.
  6. Control Ranges: Manufacturer target mean values are initially imported to the LH500 when a new lot number of control is being overlapped. These mean values are then evaluated and edited, if necessary, following a 3 day control overlap.
    Lab action limits for hematology controls are based on locally established system performance from no fewer than 60 consecutive trials over at least one and a half months after initial baseline studies and set at ± 3 SD for all reported analytes unless specifically stated otherwise in our written operational procedure.
    Action limits are set in the LH500 by entering the “set range” into the Lab limits fields for each analyte on each control screen.
    Target means for each analyte on fresh lots of control material will be set based upon a three day overlap with active (current) control prior to putting the fresh lots into service. Action limits will be fixed against these calculated means. If the initial, calculated mean from the overlap varies more than 5% from its package insert mean, contact the office of the Technical Director for instructions before releasing the control parameter for routine Q.C.
  7. Definitive language for handling “control out” situations: When a control result is outside the posted limits, first, check the Incidence Log for evidence of imprecision in that constituent.
    1. If the Incidence Log indicates that there is evidence of imprecision, stop testing and report to a Supervisor.
    2. If there is no evidence of imprecision, repeat the control once. If the control result is acceptable, proceed with patient testing. Make an Incidence Log entry including the control results and the posted limits.
    3. If the control result is not acceptable and the control result errs in the other direction, stop testing. This indicates imprecision. There are a few maintenance steps to perform to correct the problem. Check to see if there are any flags associated with the control results. This may help to determine what maintenance may be appropriate to perform first. Proceed with the following:
      1. WBC, RBC, PLT, MCV, etc.:
        1. Perform aperture zap. (See LH500 Help menu for procedure.)
        2. Clean outside of Blood Sampling Valve (BSV).
      2. Latron Control or differential:
        1. Purge the flowcell. (See LH500 Help menu for procedure.)
        2. Flowcell unclog 1, 2 or 3
          1.  After maintenance is done, retest controls. If control results are acceptable, proceed with testing.
          2.   If controls are not acceptable, call an Instrument Specialist.
          3.   In either event, make an entry in the Incidence Log.
    4. If the control result errs in the same direction, report results to Supervisor. This indicates a drift or shift in either the control material or the analyzer. Review the Incidence Log for evidence that the control constituent has been running off its target in the same direction. At that point the supervisor assumes that the error is real and determines whether or not the error has a clinical impact.
    5. If the error does not have a clinical impact, proceed with testing and report to the office of the Technical Director. The data will be reviewed to determine if the mean should be reset.
    6. If the error does have clinical impact stop testing for that constituent. There are a few maintenance steps to perform to correct the problem. Proceed with the following:
      1. WBC, RBC, PLT, MCV, etc.:
        1. Perform aperture zap
        2. Clean outside of BSV.
      2. Latron Control or differential:
        1. Purge the flowcell
        2. Flowcell unclog 1, 2 or 3
    7. After maintenance is done, retest controls. If control results are acceptable, proceed with testing.
    8. If controls are not acceptable, call an Instrument Specialist.
    9. In either event, make an entry in the Incidence Log.

Analyte Mode Match Limits

      Analyte

      Limits*

      WBC

      0.4

      RBC

      0.11

      HGB

      0.4

      MCV

      1.5

      RDW

      0.5

      MPV

      0.5

      Platelet Count

      23

      *Limited by the forced significant digit.

XXI. Reference:

  1. LH500 Operate Instructions. Beckman/Coulter.
  2. Q.C. for Hematology Analyzers with Two Sample Modes. The Joint Commission. 5/23/2005.
  3. Wu, Alan H.B., Tietz Clinical Guide to Laboratory Tests. 4th Ed.
  4. http://www.nlm.nih.gov/medlineplus/ency/article/003647.htm
  5. Steve Raymond, Dubuque Pathology Associates.
  1. Morse, E.E., G. Kalache, W. Germino, R. Stoddard; Annals of Clincal and Laboratory Science, Vol 11, Issue 2, pp 184-187 “Increased Electronic Mean Corpuscular Volume Induced By Marked Hyperglycemia”.

 

  1. December 2005 L. McGovern
  2. August 2006 L. McGovern (Revised: XIX.8. added)
  3. March 2007 S. Hosch, L. McGovern (Revised: III.7.E-F; VII.6.; XII.7, 11.A.b., 13.E.; XIII.12.E.; XIX.5.Note.)
  4. September 2007 L. McGovern (Revised: XIX.1.B.; XX.2.)
  5. October 2007 L. McGovern (Revised: XIX.1.B.d.)
  6. October 2007 L. McGovern (Revised:XIX.5)
  7. August 2009 L. McGovern (Revised: IV.; XVII.)
  8. November 2009 L. McGovern (Revised: XIX.1.A.,B., 6.)
  9. July 2010 L. McGovern (Revised: XVII.; XVIII.1-2.; XIX; align ref rngs w/ ref materials)
  10. December 2010 L. McGovern, P. Ellerbeck (Revised: XIX.plt ct; XXI.4.)

 

Comprehensive Review:

Instrument Specialist:

Technical Director:

 

Interim Review:

February 2010 K. Gourley (Revised: XII.11.A.c.; XV.2.,4.; XIX.8.C.a.1-2.,b.1)
November 2010 S. Wallace (Revised: V.1.)
December 2011 L. Begle/D. Stence/L. McGovern (Revised: XI.12-13.; XIII.11.added; XVI.13; XVIII.2., 3.C-F.; XXI.5-6.)
April 2012 L. McGovern (Revised: XVIII.2. reportable ranges; format)


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