COULTER LH 500 Shutdown and Startup Summary

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Shutdown and Startup Summary Shutdown (Cleaning Cycle) Select Diluter Functions Clean Cycle from the Instrument Computer display to bring the enzymatic cleaning agent into the Diluter. Shutdown must be performed once every 24 hours that the instrument is in use. The cleaning agent remains in the Diluter for 30 minutes. The Startup cycle begins automatically. Startup (Daily Checks) The instrument automatically performs the following checks: Background counts (CBC, DIFF) Various voltage, pressure and temperature checks Verify the Daily Checks results: Look at the Workstation Daily Checks screen (click on the Command Center, then click from the common toolbar) or look at the automatic printout of results. If any checks show FAIL then: If background, select Background from Process Type and select Start If any other checks fail, Perform a System Test Select Diagnostics Operator Options System Test from the Instrument Computer. Professional Development 1 of 18

Special HELP Icons These icons are found in the HELP system on the Workstation. Help button on Main Command Center or a screen s tool bar Help Mode (Context Sensitive Help) button In a HELP topic, shows that the topic has been changed in the current revision of the software and HELP. In a HELP topic, shows that the topic is new in the current revision of software and HELP. In a HELP topic, shows that you can find a helpful tip for a procedure by clicking on the Tip icon. Shows that you can find a list of topics related to the current HELP topic on the screen. Indicates that you can find a sub-procedure with a graphic by clicking on the Picture icon. Indicates that you can go to a video clip of maintenance or other procedure by clicking on the Projector icon. Professional Development 2 of 18

Running Controls Summary COULTER LATRON Primer and Control This control is run once per 24 hours. LATRON Primer and Control are always run as a pair. Ensure that the Primer and Control are at room temperature. The Primer is run first to clean the pathway through the flow cell. The Control is run to check and verify the VCS gains, CVs and laser alignment At the Workstation: Select Stop Choose CONTROL as the Process Type on the Command Center. Choose Ltx Primer as the Run Type Select Start Open the LATRON folder on the QC screen. Immerse the manual aspiration tip in the Primer vial, press and release the aspiration bar and remove vial when you hear the beep. If the results are #500, continue with the Control. If the results are >500, repeat the Primer. At the Workstation: Select Stop Choose Latex as the Run Type Select Start Very gently, invert the Control vial 5-8 times, being careful to avoid bubbles or foam. Immerse the manual aspiration tip in the Control vial, press and release the aspiration bar and remove vial when you hear the beep. If any results are out of range, repeat the Primer. Then repeat the Control. Professional Development 3 of 18

Running Controls Summary COULTER 5C Cell Control The frequency of running these controls is dependent upon regulatory and laboratory policy for quality assurance programs. Control Preparation: Allow vials to equilibrate at room temperature 10-15 minutes. Mix each vial using the 8 x 8 x 8 mixing method. 5C Cell Control vials receive two mixing sets. Visually check each vial after mixing to ensure cells are suspended. If cells are not suspended, then repeat one more set. 5C Cell Control Select Stop. At the Workstation Command Center select AUTO ANALYSIS as the Process Type, CD as the Run Type and Auto as the Asp Mode. Select Start. Place the control vials, barcode face up, into a regular cassette and place the cassette into the loading bay. When analysis is complete, remove control vials from the cassette and store them in the refrigerator. Review control results according to your laboratory s policy. Professional Development 4 of 18

Running Controls Summary COULTER RETIC-C Cell Control The frequency of running these controls is dependent upon regulatory and laboratory policy for quality assurance programs. Control Preparation: Allow vials to equilibrate at room temperature 10-15 minutes. Mix each vial using the 8 x 8 x 8 mixing method. RETIC-C Cell Control vials receive one mixing set. Visually check each vial after mixing to ensure cells are suspended. If cells are not suspended, then repeat one more set. Retic-C Cell Control Label two 12 x 75 ml glass vials for each control level. Pipette 50 μl of well-mixed control material into its labeled vial. Add four drops of Reagent A immediately and shake vial to mix blood and stain. Allow to incubate at room temperature for a minimum of 5 minutes and a maximum of 60 minutes. Prepare instrument to run the Retic-C control: Select Stop. Choose Process Type CONTROL and Run type R. Be prepared to type the 8-digit control lot number. Select Start. Gently remix a control/stain vial. Pipette 2 μl of the mixture into the bottom of a clean, labeled 12 x 75 ml glass vial for that control. Immediately dispense 2 ml of Reagent B into the vial, holding the vial at an angle while doing so. This ensures mixing of the contents without forming bubbles. DO NOT SHAKE. Wait 30 seconds and then analyze the retic preparation you just made. While waiting, type in the 8-digit lot number. Immerse aspirator tip in the prepared sample. Press and release activation bar (it will aspirate the entire contents). Remove the vial when you hear the beep. Review results. Professional Development 5 of 18

Calibration Summary Note: to be used by Beckman Coulter trained personnel only Calibration Procedure Prepare one S-CAL Calibrator tube according to package directions. Run S-CAL Calibrator Set up Workstation screen (remember color match) and delete previous results from the Calibration table. Set up S-CAL Calibrator reference values. Remember to choose correct S-CAL Calibrator lot# from box on left of Calibration screen. At Command Center, verify Blood Detector ON, Run Type is C, Process Type is Calibration and set # aspirations/tube to 11. After calibration runs are complete, transmit new cal factors if necessary: Select Adjust Calibration button from Workstation screen. When finished, be sure to return Workstation to your lab s default settings Rerun 5C Cell Control, if needed to verify any changes made. Professional Development 6 of 18

What To Do If Your Host/LIS Goes Down Setting up the Workstation 1. At the Workstation, from the Command Center, go to (System Set Up) (General Settings) 2. Select Cass/Pos as the only Positive Identifier if you do not have bar code labels. Select Sample ID as the only Positive Identifier if you are using bar code labels. 3. Click on (OK) to save. 4. Go to (Patient Results) (Database / ToDo List) 5. On the Common Toolbar, select (Add Sample Requests) NOTE: Before adding entries on the ToDo List, do you want to set up the Cass/Pos # to Autosequence? This makes preassigning more efficient. If YES, go to step 6. If NO, go to Processing Preassigned Samples. 6. On the Add Sample Requests window, select (AutoSequence) NOTE: Select HELP from the Setting Up AutoSequencing window to learn more about available options. 7. Set AutoSequence Control At Test Level. 8. Select to autosequence only Cass/Pos for the test modes you are running. 9. Enter the starting Cass/Pos number for each test mode. 10. Select (OK) to continue. Professional Development 7 of 18

What To Do If Your Host/LIS Goes Down Creating the ToDo List 1. On the Add Sample Request window, select Diff (for CBC/Diff), CBC (for CBC only), for example. No tube bar code labels 2. If you said Yes to Autosequencing of Cass/Pos #, you should see the first Cass/Pos # in the Cass/Pos field. 3. If you said No to Autosequencing of Cass/Pos#, type the first Cass/Pos# into the Cass/Pos field. Tube bar code labels 2. Place the cursor in the Sample ID field. 3. Use the hand-held bar code scanner to enter the Sample ID.in the Sample ID field. 4. Use the <Tab> key to move the cursor to the Patient ID field. Patient ID is the unique identifier for each sample. You must assign a Patient ID when you create a ToDo List. This can be an accession #, Medical Records #, Social Security #, -- any number that is unique to a particular patient. 5. [Optional] You may enter additional demographic information, including Patient name, Age, Gender, Physician and/or Location. 6. Select to save an entry and make additional entries on the ToDo List. 7. After making your final entry, select again. 8. Select (Close) to exit the Add Sample Request window. 9. Go to the ToDo List CBC/Diff (or other Test Mode) to verify that your entries are correct. NOTE: If the entries are not immediately visible, you may need to double click on the folder for the appropriate Test Mode to refresh the screen. 10. Remember to check the following! In (Run Configuration) Select options according to your lab protocol While the LIS/HIS is down, you may want to disable it. Remember to change your settings when the LIS/HIS comes back up. Select (SlideMaker) options, if necessary. Select (OK) to save. Professional Development 8 of 18

Front View Module Location Analyzer Module CBC Module* Electronic Power Supply BSV Module Diluter Autoloader Module Pump Module Diagram A * The appearance of some parts may be different on your instrument. Professional Development 9 of 18

Rear View Module Location Fans Reagent manifold and connectors Power ON/OFF rocker switch Plugs for computer Pneumatic Power Supply Diagram B Professional Development 10 of 18

Side View Module Location Main Diluter Module Mixing Module Flow Cell Module Diagram C Professional Development 11 of 18

Autoloader Module Unload stack full sensor Barcode reader Tube ram Load stack empty sensor Unload gate Load gate Position detect sensors Position detect sensors Unload elevator Load elevator Tube Forward Sensor Needle Assembly Tube available sensor Shown without the Needle Shield in place. Diagram D Professional Development 12 of 18

CBC Module * RBC Vacuum Isolator Chamber WBC Vacuum Isolator Chamber CBC Lytic reagent pumps (1.0 ml total) Waste Chamber RBC Aperture Bath WBC Aperture Bath Overflow Chamber Hemoglobin pre-amp Hemoglobin Lamp Diagram E * The appearance of some parts may be different on your instrument. Professional Development 13 of 18

BSV Module Manual Mode Aspiration Pump 125 μl Bubble-maker pinch valves (2) Sheath Tank Air Pump Automatic Mode Aspiration Pump 185 μl Blood Detector (rear) Blood Detector (front) Blood Sampling Valve (BSV) Probe Wipe Diagram F Professional Development 14 of 18

Pump Module Erythrolyse II Reagent Pumps (2) 300 μl (total) Diagram G Professional Development 15 of 18

Mixing Module StabiLyse Reagent Pump 133 μl Mixing Chamber Diagram H Professional Development 16 of 18

Flow Cell Module VCS Electronics Flow Cell Laser Diagram I Professional Development 17 of 18

Main Diluter Module Low Vacuum regulator Solenoids (example of one set) WBC Diluent dispense pump 6 ml 30 psi regulator RBC Diluent dispense pump 10 ml Backwash Pump 3 ml Vacuum Trap Sheath Pressure regulator* Sample Pressure regulator* *Do not adjust the Sheath or Sample Pressure unless directed to do so by your Beckman Coulter Representative. Diagram J Professional Development 18 of 18