EASY REFERENCE GUIDE

PN A86683AA (October 2009) Miami Education Center

COULTER® LH 700 SERIES

HEMATOLOGY ANALYZER

with optional

LH SLIDEMAKER AND LH SLIDESTAINER

EASY REFERENCE GUIDE


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Safety Notice Read all product manuals and consult with Beckman Coulter-trained personnel before attempting to operate instrument. Do not attempt to perform any procedure before carefully reading all instructions. Always follow product labeling and manufacturer’s recommendations. If in doubt as to how to proceed in any situation, contact your Beckman Coulter representative. Beckman Coulter, Inc. urges its customers to comply with all national health and safety standards such as the use of barrier protection. This may include, but is not limited to, protective eyewear, gloves, and suitable laboratory attire when operating or maintaining this or any other automated laboratory analyzer.

Alerts for Warning and Caution

Throughout this manual, you will see the appearance of these alerts for Warning and Caution conditions:

WARNING

WARNING indicates a potentially hazardous situation, which, if not avoided, could result in death or serious injury. May be used to indicate the possibility of erroneous data that could result in an incorrect diagnosis.

CAUTION

CAUTION indicates a potentially hazardous situation, which, if not avoided, may result in minor or moderate injury. It may also be used to alert against unsafe practices. May be used to indicate the possibility of erroneous data that could result in an incorrect diagnosis.

Safety Precautions WARNING

Risk of operator injury if: � All doors covers and panels are not closed and secured in place prior to and during instrument

operation. � The integrity of safety interlocks and sensors is compromised. � Instrument alarms and error messages are not acknowledged and acted upon. � You contact moving parts. � You mishandle broken parts. � Doors, covers and panels are not opened, closed, removed and/or replaced with care. � Improper tools are used for troubleshooting.

To avoid injury: � Keep doors, covers and panels closed and secured in place while the instrument is in use. � Take full advantage of the safety features of the instrument. � Acknowledge and act upon instrument alarms and error messages. � Keep away from moving parts. � Report any broken parts to your Beckman Coulter Representative. � Open/remove and close/replace doors, covers and panels with care. � Use the proper tools when troubleshooting.

CAUTION

System integrity could be compromised and operational failures could occur if: � This equipment is used in a manner other than specified. Operate the instrument as instructed in

the product manuals. � You introduce software that is not authorized by Beckman Coulter into your computer. Only

operate your system’s software with software authorized by Beckman Coulter. � You install software that is not an original copyrighted version. Only use software that is an

original copyrighted version to prevent virus contamination.

CAUTION

If you purchased this product from anyone other than Beckman Coulter or an authorized Beckman Coulter distributor, and, it is not presently under a Beckman Coulter service maintenance agreement, Beckman Coulter cannot guarantee that the product is fitted with the most current mandatory engineering revisions or that you will receive the most current information bulletins concerning the product. If you purchased this product from a third party and would like further information concerning this topic, call your Beckman Coulter Representative.

This document is not intended to replace the information in your instrument Instructions for

Use manual. Information in the Instructions for Use manual supersedes information in any

other manual.


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TRADEMARKS

5C, COULTER, COULTER COUNTER, FACULTY, ISOTON, LIN-C, LYSE S, and S-CAL are

trademarks of Beckman Coulter, Inc.

All other trademarks, service marks, products or services are trademarks or registered trademarks

of their respective holders.

Copyright © Beckman Coulter, Inc. 2007

All Rights Reserved


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REVISION STATUS

Revision AA Initial Issue (October 2009)

Combine the LH 750 PNA48907and LH 780 PNA36293 Easy Reference Guides.

Software 2D1/2B1 or higher

© 2009 Beckman Coulter, Inc.


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TABLE OF CONTENTS

Hematopoiesis: The Original Family Tree Normal Histograms – LH 780 and LH 750 Normal Data Plots (2D and 3D)

Workstation

Log Off/Log On LH Workstation The Command Center The Eight LH Application Buttons Special Help Icons LH SlideStainer Icons LH Workstation (computer) Shutdown

Daily Functions

LH 700 Series Analyzer CRT and Numeric Keypad Shutdown Cycle Summary Startup Cycle Summary Running Latron Summary Running COULTER

® 5C® and Retic-C™ Cell Controls Summary

LH SlideMaker SM Keypad Display Map Suggested Daily Setup for LH SlideMaker

LH SlideStainer Suggested Daily Setup for LH SlideMaker with LH SlideStainer

Calibration

Reproducibility and Carryover

Host/LIS Down

Setting Up the LH Workstation Creating the ToDo List

IQAP/eIQAP Information

General Information IQAP Download Procedure How to Register and Enroll in eIQAP


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How to Access eIQAP Upload File Messages

Parts Diagrams

LH 700 Series Diluter Front Panels Diluter Front Cover (top view)

LH SlideMaker Sample Access and Reservoir Module Dispense Module Dispense Probe Mechanism and Rinse Block Slide Transport Module

Common Troubleshooting Fixes

Common Orderable Parts LH 700 Series

Instrument Power Off/On Low Vacuum Low Message Low Vacuum Drifted Message Fluid Leaks Level Sense Message Backwash Tank Not Full Message Probe Messages Rocker Bed Messages Aspiration Messages No Diff Results No Retic Results

LH SlideMaker Dispense Probe Messages Low Vacuum 1 or 2 Messages Shuttle or Truck Vacuum Messages

LH SlideStainer Bath Not Draining Messages Bath Not Filling Messages

As Needed Cleaning Procedures

LH 700 Series LH SlideMaker LH SlideStainer

Optimizing Stain Protocols


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Hematopoiesis: The Original Family Tree

Courtesy Johns Hopkins University Press,

The Johns Hopkins Atlas of Human Functional Anatomy, Leon Schlossberg, illustrator


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Normal Histograms – LH 780 RBC Histogram Platelet Histogram WBC Histogram

Range Indicators Upper and lower limits defined by the lab or

can be turned off.

Normally has two curves. Green curve is the smooth or raw data. Black curve is

the fitted data.

Black line indicates the 35 fL lower threshold. Area checked for

presence of interference.


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Normal Histograms – LH 750

RBC Histogram

Platelet Histogram WBC Histogram

Normally has two curves. Green curve is the smooth or raw data. Black curve is the fitted data.

Black line indicates the 35 fL lower threshold. Area checked for presence of interference.


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Normal Data Plots 2D Data Plot (WBC Differential) 2D Data Plot (WBC Differential) (Lymphs removed)

Monocytes

Lymphocytes

Non-white cells

Neutrophils

Eosinophils

Basophils

Basophils


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3D Data Plots (WBC Differential) 3D Data Plot (WBC Differential) (default view) 3D Data Plot (WBC Differential) (rotated 90 degrees)

Point of origin (0, 0, 0) xyz coordinate

y-axis

Volume

z-axis Conductivity

x-axis

Light Scatter



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2D and 3D Data Plots (Reticulocytes) 2D Data Plot (Reticulocytes) 3D Data Plot (Reticulocytes)

Mature RBCs

Platelets/ Non-red cells

Reticulocytes

WBCs



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Log Off/Log On LH Workstation

To Log Off the LH Workstation

Step Action

1 Select

from the Command Center. The Log Off the System window

appears.

The default selection is Log Off Current User.

2 Select

to confirm that you want to log off the current user name.

To Log On to the LH Workstation

Step Action

Type your user name or verify the current name displayed.

1 ■ User names are defined by those with Lab Admin (Level 3) access.

Type your password.

2 Remember that passwords are case-sensitive.

If you forget your password, the Lab Admin access can reset it for you.

3 Select

at the bottom of the logon screen. The Workstation checks your

password and starts the LH Workstation.


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The Command Center

The Command Center is the green bar located across the bottom of the desktop area.

When you want to change the area you are viewing, just select a new button from the Command Center. The program automatically closes what was open and then opens the new selection.

There are three exceptions:

If the Run Configuration screen is open, you exit by selecting

either

(OK, save and close) or the red to cancel any changes

you made.

If the Help screen is open, you exit by selecting the small x in the upper

right corner of the Help screen.

If the History Logs are open, exit by selecting the small x in the upper

right corner of the History Logs screen.

The user currently

logged on.

Body Fluid Option

Eight LH

Application buttons

Status Indicators for Analyzer or SlideMaker and

PC

Barcode entry Scanned or Typed

Predilute Optionwith

dilution factor entry

Process Type

Usually has Auto

Analysis selected.

Default Type

Selects the sample test mode if no barcode or

Random Access

Enable/Disable

Random Access

Traffic-light iconInstrument Status


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The Eight LH Application Buttons

Run Configuration

Takes you to the screen to view/change Print, LIS and LH SlideMaker run choices. Also access to AutoStop choices.

Patient Results

Takes you to the screen to view patient results from either the Results/Graphics view or the Database/ToDo view.

Quality Assurance

Takes you to the screens associated with various QA data: Daily Checks, Quality Control, Extended QC, XB, XM, Reproducibility, Carryover, Calibration.

Help

Provides access to an extensive multimedia help system built into the software.

History Logs Viewer

Provides access to the various electronic logs for review, to add comments, print or archive.

System Status

View current details about the system , such as software revision.

System Setup

Provides access to the various setup functions to customize your workflow: General Settings, Quality Assurance, Patient etc.

Log Off (Exit) the System

Choose between Log off current (default) or Shutdown the Workstation.


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Special HELP Icons

These icons are found within 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 was changed in the current revision of the software and HELP.

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.


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COULTER®

LH SlideStainer Icons

From System Set Up, go to . Select tab.

Save Protocol

Saves newly created or modified staining protocol.

Delete Protocol

Deletes selected protocol. You cannot delete a protocol while it is in use.

Download SlideStainer Protocol (LabAdmin only)

Downloads a saved or modifiedstain protocol to the SlideStainer. The icon will only be available when the SlideStainer is in Standby mode.

New Protocol

Provides blank Staining Protocol data fields to prepare to set up a new protocol or to modify an existing protocol.

Open SlideStainer Arm Dialog Box (LabAdmin only)

Arm is aligned in response to error situations.

SlideStainer Status

Provides information about Serial # and cycle count among others. Buttons to open and close gripper and reinitialize arm.

SlideStainer Input Queue Status (LabAdmin only)

Displays SlideStainer input/output queue configuration for configuring the “parking lot” / empty basket areas.

Drain SlideStainer Reagent Lines

In preparation for shutting down SlideStainer or preventing spillage during maintenance procedures.

Monitor Sensor Status

To troubleshoot error messages. Also monitor values for temperatures, pressure sensors, and battery VDC information.

Download SlideStainer Software (LabAdmin only)

To upgrade software or respond to error situations


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LH Workstation (computer) Shutdown

The LH Workstation (computer) should go through this procedure at least once per week.

You may have to shut down and restart the Workstation in response to certain error

situations or if the computer doesn’t respond. Be sure to read any Help message for error

recovery carefully for this indication.

In the event of a communication problem between the Workstation and the LH 780

Analyzer, you may need to reset the Analyzer using the RESET button on the upper right

corner of the Analyzer cover as well as selecting to shut down and restart the Workstation.

Step Action

1 Select from the Command Center.

2 Select Shutdown the Workstation, then Select OK

The Workstation will save any unsaved data, close all

applications and display a window notifying you it is shutting

down the workstation. This may take a minute.

Next, a window is displayed that states you may now power

off the workstation.

3

Turn off the computer workstation power using the power

button on the front of the computer tower.

Always verify that the power actually went off. Sometimes,

you need to hold in the button until all power is off.

4 After one minute, turn the power back on.

5 Log on to the workstation when the Log On window

reappears.


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Analyzer CRT and Numeric Keypad

LH 780

LCD Display

Numeric Keypad

Always return to this screen to run the instrument.

Use this button to access the Main Menu


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Analyzer CRT and Numeric Keypad

LH 750

LCD Display

Numeric Keypad

Always return to this screen to run the

instrument.

Use this button to

access the Main Menu


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Shutdown Cycle Summary

Shutdown (Cleaning Cycle)

Step Action

1 Verify pneumatics are On. (Look at the Power Supply display. If off, press the key on the Analyzer Numeric Keypad.)

2a If you have an LH SlideMaker, select Shutdown from the SlideMaker keypad menu. Wait until cycle completes before proceeding to next step.

2b

When the Analyzer CRT and the LCD display both indicate “Ready”, press the key on the Analyzer Numeric Keypad. This pulls the enzymatic cleaning agent into the Diluter.

� Shutdown must be performed once every 24 hours

that the instrument is in use.

� The cleaning agent must remain in the Diluter for a

minimum of 30 minutes.

Note � Cleaning agent removal cycles are performed automatically at the end of the programmed cleaning time. (30 minutes to 24 hours)

� No Background Checks or Electronic Tests are

performed at this time.

3 Proceed to Startup Summary


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Startup Cycle Summary

Startup (Daily Checks)

Step Action

1 Press the key on the Analyzer Numeric Keypad.

� After the cleaning agent removal cycles, this is the

only key you can use.

The instrument automatically performs the following checks: � Background counts (CBC, DIFF, RETIC) � Ramp & Precision electronic checks � Various voltage, pressure and temperature checks

Note If you have an LH SlideMaker, it will startup automatically if it was in Shutdown. If not, run Startup from the SM keypad.

2

Review 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 background or main system checks show FAIL then repeat the required test from the Analyzer CRT screen:

� Select Main Menu, Analyzer Functions, Startup Tests.

� As required select Background or Ramp or Precision.


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Running Latron Summary

COULTER® LATRON™ Primer and Control

� Run this check once every 24 hours.

� LATRON Primer and Control - always run as a pair.

� Always run the Patron Primer first to clean the pathway through the flowcell.

� Follow with the Latron Control to check and verify the VCS gains, CVs and laser alignment.

Step Action

1

At the Workstation:

� Choose Auto Analysis in the Process Type field on the Command Center.

� Open the LATRON folder on the QC screen.

2 At the Analyzer Numeric Keypad, press (Diff & Retic) or (Diff). Then press .

3

Press the key on the Analyzer Numeric Keypad to run the Primer.

� Present the Primer vial to the manual aspiration tip and remove when you hear the beep.

� If the results are ≤ 500, continue with the Control.

� If the results are > 500, repeat the Primer.

4

Press to continue (do not press the key again)

� Very gently, invert the Control vial 5-8 times, being careful to avoid bubbles or foam.

� Present the Control vial to the manual aspiration tip and remove when you hear the beep.

� If any results are out of range, repeat the Primer, then repeat the Control.

5 Press the key twice to return to READY at the

Analyzer Numeric Keypad.


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Running 5C and Retic-C Cell Controls Summary

COULTER® 5C® Cell Control and RETIC-C™ Cell Control

The frequency of running these controls depends upon regulatory and laboratory policy for quality assurance programs.

Step Action

1 Allow vials to equilibrate at room temperature for 10-15 minutes.

Mix each vial individually, by hand, using the 8 x 8 x 8 mixing method as described in the control package insert:

a) Begin with the vial upright between the palms of your hands and roll the full length, forward and backwards eight times. (first eight)

b) Next turn vial upside down between the palms of your hands and roll the full length, forward and backwards eight times. (second eight)

2

c) Next hold vial in one hand and invert eight times. (third eight)

Note

1

Perform the above steps twice for 5C Cell Control vials.

For RETIC-C Cell Control vials perform the above steps once.

3 Visually check each vial after mixing to ensure cells are suspended. If cells are not suspended, then repeat one more set.

4 Place the control vials, barcode face up, into a regular cassette and place the cassette into the loading bay.

5 When analysis is complete, remove control vials from the cassette and store them in the refrigerator.

Note

2

The entire process, steps 1-5 must be completed within 30 minutes to preserve control stability.

6 Review control results according to your laboratory’s policy.


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COULTER® LH SlideMaker Keypad Display Map

MAIN MENU

ROUTINE FUNCTIONS*

SERVICE FUNCTIONS

SPECIAL FUNCTIONS

RUN MODE

SPECIAL CYCLES

MAIN MENU – ROUTINE FUNCTIONS*

PRINTER LABELFUNCTIONS

Save Printer Label IntensityIncrement Intensity

Decrement Intensity

SYSTEM SETTINGSDisplays Software/Hardware Rev

ROUTINE FLUIDICS*

PNEUMATIC/POWER SUPPLIES

Displays Pressures, Vacuums, Voltages

ADVANCE BASKET

Manually advance basket one position

UNLOCK CASSETTE Manually unlocks slide cassette

LOCK CASSETTE Manually locks slide cassette

Prime Reagent Reservoirs

Drain

Run STARTUP*

Backwash Tubing

Drain Vac Accumulator

Run SHUTDOWN*

MAIN MENU – SERVICE FUNCTIONS Service Use Only – needs Service Password

MAIN MENU – SPECIAL FUNCTIONS

DEFINE SMEAR SPEED

DRYER TEMP

DISPLAY TEST

DRYER FUNCTION TEST

REBOOT PRINTER

SENSOR STATUS

KEYPAD TEST

MAIN MENU – RUN MODE

VIEW ERROR

MAIN MENU – SPECIAL CYCLES PRINT LABEL

Prints test “Dummy” label after replacing Labels or Ribbon

BLEACH SYSTEM

• The choices in green are your daily functions.


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LH SlideMaker Suggested Daily Setup

These checks/setups may be done during the Shutdown cycle, or at any time appropriate for your lab.

LH SlideMaker - only

� Verify that the Slide Cassette is full.

� Check Label and Ribbon supply, replace if necessary. (refer to Help)

� Place up to twelve empty slide baskets in the front and rear tracks. (if no LH SlideStainer)

� Inspect dispense probe and rinse cup and clean if any traces of blood are seen. (refer to Help)

Once the system is operating automatically, do not grab slides prematurely, this can cause jams and resets. Let the SlideMaker do its job.


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LH SlideMaker With LH SlideStainer Suggested Daily Setup

These checks/setups may be done during the Shutdown cycle, or at any time appropriate for your lab. Depending on workload, SlideStainer baths may need attention more than once a day.

LH SlideMaker

� Verify that the Slide Cassette is full.

� Check Label and Ribbon supply, replace if necessary.

�

Verify there are no more than six slide baskets in the SlideMaker and locate them in the rear track. There should be no baskets in the front track.

� Inspect dispense probe and rinse cup and clean if any

traces of blood are seen. (refer to Help)

LH SlideStainer

� Place the SlideStainer in Standby state.

� Place six slide baskets in the Empty Basket area.

� Verify bath reagent container levels. Replace/Refill as necessary.

� Drain all baths.

� Check Waste Containers. Empty if necessary.

(Waste 1 = Aqueous, Waste 2 = Methanol)

� Check the “stain” baths for excess sediment and clean with methanol if necessary.

� Fill all baths (according to your lab’s protocol)

�

Place the SlideStainer in Auto state. Once the major workload finishes, empty baskets are stained if left in Auto. You may change between Standby and Auto state.

Once the system is operating automatically, do not grab slides prematurely, this can cause jams and resets. Let the SlideMaker and SlideStainer do their jobs.

Note: Your Applications Specialist may help you to revise

these setups to better suit your lab’s protocols and needs.


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Calibration Summary

Note: to be used by Beckman Coulter trained personnel only

1 Shutdown for 30 minutes to ensure apertures are clean.

2

Check Reagent Levels:

� Diluent (1/2 to 2/3 full)

� CBC Lytic Reagent (¼ full – 1L or 5L)

3 Run Startup and verify Daily Checks results.

4 Analyze COULTER®5C® Cell Controls.

5 Disable Automatic Print on the Run Configuration screen.

6 Prepare one S-CAL® Calibrator tube according to the package insert directions.

7

Run S-CAL Calibrator:

� Set up Workstation screen as described in HELP and delete previous results from the Calibration table.

� Set up S-CAL Calibrator reference values required for your reagent system.

� Remember to choose the correct S-CAL Calibrator lot # from the box on the left of the Calibration screen.

� At the Analyzer CRT screen, turn Blood Detector ON, set the # of aspirations/tube to 11, verify Default Type is C.

8

After calibration runs are complete, review the calibration data and transmit new cal factors if necessary:

� Prepare Analyzer to RECEIVE new values:

� From the Analyzer CRT screen, select the following: Main Menu, Analyzer Functions, Calibration, Receive Average Cal Factors.

� Select Adjust Calibration button on the specific toolbar from the Workstation calibration run screen.

9 When finished, be sure to return the Workstation and Analyzer CRT screen to your lab’s default settings.

10 If any calibration factor changes were made, rerun 5C Cell controls to verify the calibration.


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Reproducibility and Carryover Summary

Note: to be used by Beckman Coulter trained personnel only

Disable Automatic Print on the Run Configuration screen.

1 Run Reproducibility (Default Type set to C or CD) with the selected normal whole blood specimen. Refer to HELP for sample requirements.

a Set up the Workstation screen as described in HELP and delete previous results from the table.

b Set the # aspirations/tube to 1 and cycle a separate sample collected in a salt of EDTA.

c Using 2-6 tubes from the same donor (5 mL EDTA), pool, mix and aliquot the sample into 2-6 clean EDTA tubes.

d Set the # of aspirations/tube to 5 on the Analyzer CRT screen.

e Place 2 of the tubes in the same cassette in the right loading bay.

f After the samples are analyzed, review the results.

Note 1 The procedure begins with an N=10. If all parameters pass, you may stop processing. Otherwise, you may need more data. See Note 2 below.

g Verify that the %CV (Coefficient of Variation) does not exceed the established limits.

Note 2

Reproducibility Limits for CBC

Parameter %CV

WBC ≤ 1.7

RBC ≤ 0.8

Hgb ≤ 0.8

MCV ≤ 0.8

Plt ≤ 3.3

MPV ≤ 2.2

If any limit is exceeded, repeat with two more tubes of the prepared sample blood, until at least 11 but not more than 31 whole blood samples have been analyzed. You may stop the analysis at any point between 11 and 31 runs if the reproducibility established limits have been met. You may not pick and choose which runs are included. Runs included for the assessment need to be consecutive runs.

If any instrument results exceed these limits, repeat with another donor sample. If you still have a failure, an instrument problem may exist. Call your Beckman Coulter Representative. (Note: the above limits are also available in HELP)


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2 Run Carryover study (Default Type set to C) with the selected blood samples and 3 separate diluent tubes.

a Set up Workstation screen as described in HELP and delete previous results from the table.

b Set # aspirations/tube to 1 and turn the Blood Detector OFF at

the Analyzer CRT screen.

c Run the samples and diluent tubes.

d Check results.


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What To Do If Your Host/LIS Goes Down

Setting up the LH Workstation

1

At the LH Workstation, from the Command Center, go to:

(System Set Up)

(General Settings)

2 Select Cass/Pos or Sample ID as applicable to your situation:

a Select Cass/Pos as the only Positive Identifier if you do not have bar code labels.

b 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.


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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.

Tubes without bar code labels Tubes with 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.

2

Place the cursor in the Sample ID

field.

3

If you said No to Autosequencing of Cass/Pos#, type the first Cass/Pos# into the Cass/Pos field.

3

Use the hand-held bar code scanner to scan the Sample ID in the Sample ID field. Verify ID scanned.

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 any number that is unique to the patient, e.g., accession #, Medical Records #, Social Security #.

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 Host/LIS is down, you may want to disable it. Remember to change your settings when the Host/LIS comes back on-line.

� Select (SlideMaker) options, if necessary.

� Select (OK) to save.

NOTE

It is also possible to print two sample reports during a down time, one Chartable and one Laboratory. Go to System Setup>>General>>Reporting Options>>Printout Options. Save. Then go to Run Configuration and set print to All Samples. Save.


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IQAP/eIQAP General Information

Important Points to Remember When Setting Up eIQAP

Make note of the following:

Instrument IQAP Participant # Serial Number System ID

Number to Call for Help: 800-526-7694 (USA and Canada)


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IQAP Download Procedure

Step Action

1 Select on the Command Center.

2 Select to display the CONTROLS window.

3 Select . An IQAP Disk Storage window appears.

4 Ensure the removable media (formatted CD, flash drive, or floppy disk) is inserted in the appropriate drive or connected to the computer.

5 Select the lot numbers you want copied to the removable media.

6 Select to begin copying the control information.

7 The "Save as" window appears.

8 Specify the location of your removable media and select "Save".

9 After the download process is complete, retrieve the removable media from the appropriate drive or USB location

10 Use the files downloaded to submit the data through the eIQAP website. Files may also be submitted via e-mail or as a last resort mailed to IQAP.


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How to Register and Enroll in eIQAP You must be registered on the Beckman Coulter website to gain access to the Electronic Quality Assurance Programs. In order to access the Hematology eIQAP program, you need to set up your account (i.e. obtain your instrument’s IQAP number).

How to Access eIQAP

Step Action

1 Go to www.beckmancoulter.com

2 Hover over the words “Customer Support” on the menu bar.

3 From the drop down box choose “QA & Service Programs”

4 Select the link “Enroll in IQAP” near the middle of the page

5 On the Quality Assurance Programs window there are links to register and enroll.

Note If you already have a login ID and password, proceed to "Enroll in eIQAP". Skip the registration.

Step Action

1 Go to www.beckmancoulter.com .

2 Hover over the words “Customer Support” on the menu bar.

3 From the drop-down box choose “QA & Service Programs”.

4 Select the link “Go to eIQAP” near the middle of the page.

5 On the Quality Assurance Programs window, select the link “Go to Hematology eIQAP”..

6 On the next window enter your Beckman Coulter username and password.

7 On the following page look for Hematology Resources and select “eIQAP Help”. This begins a breeze presentation designed to walk you through the functionality provided in the Hematology eIQAP application.


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Upload File Messages

Message Interpretation

Upload successful The files have been successfully uploaded.

ID on diskette doesn’t match instrument’s ID

The IQAP ID on the diskette, obtained from the instrument setup, does not match the IQAP ID displayed for the selected instrument.

1. Ensure the data on the diskette is from the selected instrument.

2. Ensure the IQAP ID is entered correctly in the instrument setup.

Not matching serial number on diskette

The diskette serial number, obtained from the instrument setup, does not match the serial number displayed for the selected instrument.

1. Ensure the data is from the selected instrument. 2. Ensure the serial number is entered correctly in the

instrument setup. 3. Ensure there are no spaces or symbols within the serial

number.

Drive not ready or unreadable diskette

The drive cannot access or read the diskette. 1. Ensure the diskette is inserted into drive A. 2. Repeat download and upload process with a new diskette. 3. Ensure diskette drive is functional.

Windows temporary folder missing

The Windows operating system temporary folder is missing.

Invalid data collection software

Coulter STKS, MAXM, or HMX data was collected with an unacceptable software version.

1. Reconfigure diskette at the Resources page. 2. Repeat download and upload process with newly

configured diskette.

Not an IQAP diskette or incomplete download

Diskette is not configured as an IQAP diskette. 1. Ensure diskette is from IQAP download. 2. Ensure instrument provided a successful download

message. 3. Repeat download and upload process with a new diskette.


25 of 59

Message Interpretation

Incomplete control data file

The diskette does not contain a complete control file. 1. Ensure instrument provided a successful download

message. 2. Repeat download and upload process with a new diskette.

Failed to upload IQAP files

An interruption occurred during the transfer of data through the internet.

1. Repeat upload process at a later time using the same diskette.

Insufficient privilege to access file system

Data on the diskette cannot be accessed as the applet certificate that is required to upload the IQAP files from a diskette was not accepted.

1. Logout of eIQAP and out of the Beckman Coulter website. Reenter eIQAP and accept the certificate at the Upload Files page by selecting either Yes or Always.

2. If unable to accept the certificate, enter data manually.

Drive write protected

Data on the diskette cannot be accessed as the diskette is write protected.

1. Ensure the window in the lower left corner of the diskette is closed.

Couldn’t create files in the floppy, please check it’s writable and has xxxx bytes or more free in the floppy

The diskette does not have sufficient space to create the IQAPUPLOAD file.

1. Repeat download and upload process with a new diskette.

Corrupted IQAP file on MAXM, STKS, or HmX diskette

The file has been corrupted.

1. Repeat download and upload process with a new diskette.

PN

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32 of 59

Common Troubleshooting Fixes

Common Orderable Parts

Fluidics

Low Vacuum Low Message

Low Vacuum Drifted Message

Fluid Leaks

Level Sense Message

Backwash Tank Not Full Errors

Mechanical

Probe Errors

Aspiration Errors

Rockerbed Errors

Results

No Diff Results

No Retic Results

LH SlideMaker

Dispense Probe Errors

Vacuum Errors

LH SlideStainer

Bath Not Draining Errors

Bath Not Filling Errors


33 of 59

Common Orderable Parts

Part Number Part Description Quantity P.O.6807145 Needle Pierce Cartridge Assembly 1 Ea Y

3203015 Tubing, silicone, round, 0.062 id., black stripe 10 Ft. N

3213135 Tubing, silicone, I-beam, 0.062 id., black stripe 10 Ft. N

3213163 Tubing, silicone, round, 0.062 id x .047W, brown 10 Ft. N

3203016 Tubing, silicone, round, 0.079 id, blue stripe 10 Ft. N

3213137 Tubing, silicone, I-beam, 0.079 id, blue stripe 10 Ft. N

3213176 Tubing, silicone, round, 0.079 id x .039W, yellow 10 Ft. N

6706295 Pick-up tubing, universal, 10-20L, Clenz 1 Ea Y

6706296 Pick-up tubing, for 5L Lyse S-III 1 Ea Y

6706297 Pick-up tubing, 0.5-1.9L for 1L Lyse S-III and Pak 1 Ea Y

6805636 Waste line tubing 1 Ea Y

178453 Diluent pick-up tube with float sensor, 20L 1 Ea Y

5120248 Tubing, Waste Level Sensor Assembly 1 Ea Y

1022450 Air Filer (under Power Supply) 1 Ea Y

1022868 Air Filter (rear cover Analyzer) 1 Ea Y

2517119 Screw cap for large pick-up tubes 1 Ea N

2523574 Screw cap for small pick-up tubes 1 Ea N

6232263 “Y” connector, .050 id x .130 od 2 Ea N

6232259 Fitting, hose barb, “Y”, .093 id x .093 od 2 Ea N

3202036 Tubing, polyurethane, .082 id x .144 od 14 Ft N

6855763 Pull apart std. pinch valve, single 2 Ea N

5415206 Pinch valve removal tool 1 Ea Y

6605505 Glass slides, 1 gross=2 packages SM 5-10 Gr Y

2016733 Slide Labels, 1500/roll SM 1 Roll Y

2016732 SM Printer Ribbon SM 1 Roll Y

6806470 Slide Cassette SM 1 Ea Y

6806894 Slide Basket SM 1 Ea Y

2603047 Air filter SM 1 Ea Y

1024888 Bath, Liquid ST 1 Ea Y

6807094 Reagent Rack, round opening (1 gal.) ST 1 Ea Y

6807093 Reagent Rack, 2 rectangular openings (2L)

ST 1 Ea Y

1024889 Tray, bath, plastic (holds five baths) ST 1 Ea Y

1024915 Tray, divider ST 1 Ea Y

6706296 Tubing, pickup assembly, reagent ST 1 Ea Y

6806976 Filter, Liquid, Waste / Reagent, w/tubing ST 1 Ea Y

5120249 Pickup tubing, Di water, level sense assy.

ST 1 Ea Y

A35473 Accuvette (200/bx) 1 Bx Y


34 of 59

Instrument Power Off/On

Power Off

Step Action

1 Press on the Analyzer Control keypad.

2 Turn off the main breaker switch on the left side of the Power Supply.

WARNING Risk of personal injury. A shock hazard exists if the power cord is connected. Unplug the primary power cord before performing these procedures.

3 Disconnect the primary power cord at the source (not at the power supply).

Power On

Step Action

1 Reconnect primary power cord.

2 Turn on the main breaker switch on the left side of the Power Supply

3 Press on the Analyzer Control keypad.

LH SlideMaker

There is a power button on the front of the LH SlideMaker to turn power off and on. If required by a procedure, there is a Main Power switch on the rear of the SlideMaker.

LH SlideStainer

There is a power button on the left side of the LH SlideStainer to turn power on and off. If required by a procedure, there is a Main Power switch on the rear of the SlideStainer.


35 of 59

Low Vacuum Low Message Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.

Low

Vacuum

Low?

Start Yes

1. Empty/Clean jar**.

2. Ensure float is down.

3. Reattach jar without

cross-threading.

Verify Low Vacuum

reading is 6.000 ± .010

using F92.

Adjust if necessary.

Is Vac Trap

Full?

Call BCI

Yes

YesNo

No

Done

Yes

Look at your

Vacuum Isolator

Chambers (VICs)

Yes

No

No

Are one or both

of the VICs full

of liquid?

No

Yes

Yes

Are one or both

VICs cracked?

Call BCI

Did VICs

drain?

Drain the VICs.

Press F05, then

16,

then Enter

Press Stop

three times.

Verify Low Vacuum

reading is 6.000 ± .010

using F92.


Are all lines

connected to top

of VICs?

Are there

holes in any of

the lines?

Reconnect

Replace the

tubing.

No

Verify Low Vacuum

reading is 6.000 ± .010

using F92.


Yes Yes

**Power Off instrument

before performing

procedure.

Follow HELP.

Able to

adjust?

Verify.

Run a

specimen

Verify Low Vacuum

reading is 6.000 ± .010

using F92.


YesAble to

adjust?

Flush Low Vac Line

(see attached

procedure)

Able to

adjust?

Done

No

No

No

Resolved?

Yes

No


36 of 59

Check the Vacuum Overflow Tank (VC22). If

over half full stop and call BCI. If empty or less

than half full, proceed to next step.

Remove the right side panel.

Open the lower door.

Remove the clear plastic shield.

Locate the low vacuum tubing connected to the

area labeled COUNT VACUUM on the right rear

panel. Disconnect the smaller tubing from the

front of the fitting.

Dip the disconnected end of the tubing into a

container with no more than 20 mL of DI water

until it is all aspirated.

Leave the tubing disconnected for ten (10)

minutes while it draws air to dry the vacuum line.

Reconnect the tubing, replace the plastic shield,

close the door and replace the right side panel.

Locate the Vacuum Overflow Tank (VC22) and

disconnect the float sensor connector, P-13/J-

13.

Press STOP on the keypad and allow VC22 to

drain.

Once VC22 is empty, reconnect P-13/J-13. (If

alarm is triggered, “VACUUM OVERFLOW

TANK STILL FULL” press ALARM RESET on

the keypad and disregard error message.)

Flush Low Vacuum Line

Return to

Low Vac Low

flowchart


37 of 59

Low Vacuum Drifted Message Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.

Low

Vacuum Drifted?

No

Yes

Won’t adjust? Sluggish adjustment?

Drifting?

Yes

Yes

Yes

All Pass &Message gone?

Perform Startup

Start

Verify Low Vacuumreading is 6.000 ± .010

using F92.Adjust if necessary.

Perform F92.Verify low vac

reading 6.000 ± .010

Call BCIDone

Check all count tubings. Verify tubing not cut through pinch

valves. Verify RBC sweepflow tubings secure.

Call BCI

Found issue with

tubing?

Yes

Able to adjust?

Flush Low Vac Line

(see attached procedure)

Repair and replace tubing.

Able to adjust?Verify Low Vacuum

reading is 6.000 ± .010 using F92.


No

No No

Perform Reset

Message reoccurs?

Yes

No

Yes

Run Specimen

No


38 of 59

Check the Vacuum Overflow Tank (VC22). If

over half full stop and call BCI. If empty or less

than half full, proceed to next step.

Remove the right side panel.

Open the lower door.

Remove the clear plastic shield.

Locate the low vacuum tubing connected to the

area labeled COUNT VACUUM on the right rear

panel. Disconnect the smaller tubing from the

front of the fitting.

Dip the disconnected end of the tubing into a

container with no more than 20 mL of DI water

until it is all aspirated.

Leave the tubing disconnected for ten (10)

minutes while it draws air to dry the vacuum line.

Reconnect the tubing, replace the plastic shield,

close the door and replace the right side panel.

Locate the Vacuum Overflow Tank (VC22) and

disconnect the float sensor connector, P-13/J-

13.

Press STOP on the keypad and allow VC22 to

drain.

Once VC22 is empty, reconnect P-13/J-13. (If

alarm is triggered, “VACUUM OVERFLOW

TANK STILL FULL” press ALARM RESET on

the keypad and disregard error message.)

Flush Low Vacuum Line

Return to

Low Vac Drifted

flowchart


39 of 59

Fluid Leaks Review information in HELP and in the TS3-Replacing Components Module. Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.

I-beam

tubing leak?

Round, red stripe

silicon in Random

Access Module?

Other

tubing leak?

**Find correct

replacement

I-beam tubing with

same color stripe.

Cut new piece

of tubing to

same length as

original.

Disconnect leaky

tubing* and pull

out of pinch valve.

Thread new piece

of tubing carefully

through tracks in

pinch valve.

Yes

Yes

No

**Find correct

replacement

tubing with same color

stripe or part number.

Carefully

reconnect the

two ends.

Start

Done

Cut new piece

of tubing to

same length as

original.

Disconnect leaky

tubing*.

Carefully connect

new tubingDone

No

**Find correct

replacement

tubing with same

color stripe.

Cut new piece

of tubing to

same length as

original.

Yes

No

Disconnect leaky

tubing*.

Carefully connect

new tubingDone

Carefully thread

new tubing

through blue

spacer sleeve

(Fig. 1)


before performing

procedure.

Follow HELP.

Fig. 1

*Be sure to mark

the exact places

where tubing is

removed.


40 of 59

Level Sense Message Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.

Start

Done

Call BCI

Is

container

empty?

Yes

Change container

and update

reagent log.

No

Verify all tubing

connections from

reagent to

instrument.

If diluent, check float

sensor in container

and sensor

connection (Fig. 1).

Any tubing

disconnected/

pinched?

No

Yes

Reconnect/

straighten

tubing

Reagent/

Level out

message

Press Startup

and observe.

Level out

message?No Yes

**Look at reservoirs

in back of unit.

(Open door on right

side, back)

Check the I-beam

tubing from

reservoir thru

pinch valve. (Fig. 2)

YesReplace I-

beam tubing**

No

Any bubbles

or leaks?

NoYes

Reconnect or

replace leaky

tubing

Problem

continues?

Yes

NoDone


before performing

procedure.

Follow HELP.

Perform F16 to

Prime Diluent.

Did system

prime diluent?

Replace

Diluent pickup

tube.

No

Verify.

Run a

specimen.

Yes

Level out

message?

Yes

Press Startup

and observe.

No

Retic

Clear

Retic

Stain

Pack

Prsrv

Pack

Lyse Cleaner

Fig. 2

CBC

Lyse

Fig. 2

Sensor Connector

Fig. 1

Is I-beam

tubing

crimped?


41 of 59

Backwash Tank Not Full Message Whenever performing any troubleshooting procedures, always wear all required personal protective

equipment.


42 of 59

Ensure all materials are on hand including a 20 mL

syringe with a piece of yellow-striped tubing (5-inch)

attached to the tip and some distilled water.

Remove the left diluter panel by lifting up and out on

the panel.

Locate VL-107A, the white pinch valve located

above the Backwash Tank. The top fitting above VL-

107A is a feed-through fitting marked FF-5.

Disconnect the I-beam tubing from FF-5.

Fill the syringe with distilled water and attach the

syringe tubing to FF-5.

Vacuum draws the water out of the syringe. If

needed, you may push on the plunger if the water

does not pull on its own.

The water fills the Vacuum Overflow Tank (VC 22)

Use enough water so that VC 22 is half full.

Disconnect the syringe tubing and reconnect the I-

beam tubing to FF-5.

Press the Reset button on the Analyzer. VC 22

drains.

Perform Diluter Function F-30 to activate the

Backwash Tank Check to confirm resolution.

Run a blood sample to verify Instrument

performance.

Flush Backwash Tank Vacuum Supply Line

Return to

Backwash Tank

Not Full flowchart


43 of 59

Probe Messages Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.


44 of 59

Rocker Bed Messages Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.

Start

Done

Call BCI

Any physical

obstruction?

Message:

Bed not Forward

Bed not Backward

Bed not Level

F95 – release

all solenoids

Manually rock

the bed to see

if there is any

physical

obstruction.

Remove it/

Fix it

Yes

Clean

sensors with

canned air.

No

Locate the

sensors at the

right side of the

rocker bed (see

Fig. 1 & 2)

Message

gone?

Yes

No

Start

Message:

Bed Not

Advanced

Place a sample

cassette with tubes

on the bed and

slide under tube

detector.

Any

obstruction?Call BCIYes

Perform the As-needed

Rocker Belt Procedure

Wipe belt with moist

gauze (DI water) top and

bottom. Pull belt from right

to left to clean all.

Dry thoroughly, top and

bottom.

No

Use F96 to

verify belt

movement.

Movement

OK?Done

Call BCI

Yes

No

Fig. 1 – Top view

Fig. 2 – Side view

Flag in

sensorBed Not

Backward

Bed Not

Level

Bed Not

Forward


45 of 59

Aspiration Messages

Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.

Start

Done

Call BCI

Aspiration

message and

alarm received

Check the Help

information for the

particular aspiration

message (N, B, P, C)

received

Is the message

occurring on more

than one sample?

Check the sample tube

for correct blood

volume (at least 1mL of

properly collected

blood)

No

Clean and

exercise shear

valve 85/126

Yes

Perform the as needed Clean Shear

Valves Procedure

1. Locate shear valve 85/126.

2. While watching the ceramic in the left

side, activate Sol 126. Observe the space

created between the ceramic and the

metal.

3. Clean this space with DI water and dry

with lint-free tissues.

4. Press Stop three times to exit the

solenoid function.

5. Rerun sample(s).

Check the

sample tube

for clots.

Check if Hgb

result <4 g/dL.

Proper amount

of blood?

Redraw

sample

No

YesClots found?

No

Hgb < 4 g/dL?

Rerun in

Manual

aspiration

mode

Yes

No

Rerun in

Automatic

aspiration

mode

Yes

Still aspiration

messages?Yes

NoNo

Still aspiration

messages?

YesPerform Flush

Needle/

Aspirate Pump

Pathway

(see attached

procedure)

Replace

needle

Yes

Still aspiration

messages?Yes Done


46 of 59

Read all the instructions before beginning the procedure.

Needed:

• COULTER® cleaning agent

• 10-mL syringe (PN 2523145)

• 5” (12.7 cm) yellow-striped tubing (PN 3213176)

• Cassette

• Three (3) previously analyzed samples

1. Ensure that all safety precautions are followed while servicing BCI equipment. This

includes (but is not limited to) using proper lab protocol, identifying potential blood borne

pathogen exposure, using personal protective equipment, and properly disposing of parts.

2. Power off the instrument and power supply.

3. Push the aspiration tip for the manual mode all the way back in.

4. Fill syringe with 4 mL COULTER® cleaning agent (more than 4 mL will result in spillage).

5. Connect 5” (12.7 cm) yellow-striped tubing to syringe.

6. Remove tubing at fitting FF160 (refer to FIGURE 1). FF160 is located approximately five

inches (12.7 cm) to the left of the blood sampling valve and five inches (12.7 cm) from the

front of the diluter.

7. Connect yellow-striped tubing from syringe to fitting FF160.

8. Inject the COULTER® cleaning agent into FF160. The fluid will fill needle bellows (rubber

sleeve surrounding the needle) – pressure should be minimal.

9. Exercise the syringe (push plunger back and forth) to ensure that pathway is clean.

10. Remove yellow-striped tubing from FF160 and reattach brown-striped tubing from VL-9B

to fitting FF160.

11. Power up system.

12. Perform Start-up.

13. Run bloods to verify problem resolution.

Flush Aspiration Pathway Between the Needle and Aspirate Pump

Return to Aspiration

Messages Flowchart

Fig. 1


47 of 59

No Diff Results



48 of 59

No Retic Results Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.


49 of 59

Dispense Probe Messages On LH SlideMaker

Use the diagrams for Dispense Probe Mechanism and Slide Transport Mechanism on pages 34 and 35 of this guide to help locate the sensors. Whenever performing any troubleshooting procedures, always wear all required personal protective equipment.


50 of 59

Low Vacuum 1 or 2 Messages on LH SlideMaker



51 of 59

Shuttle or Truck Vacuum Messages on LH SlideMaker



52 of 59

Bath Not Draining Messages On LH SlideStainer



53 of 59

Bath Not Filling Messages On LH SlideStainer



54 of 59


LH 700 Series

Air Filters

Aspiration Probe Wipe

BSV Drip Plate

BSV Outside

Clogged Flow Cell

Needle Assembly

Rocker Bed Belt

Stripper Plate

Shear Valves

LH SlideMaker

Air filter

Blood Spills and Broken Slides

Catch Tray

Dispense Probe and Rinse Cup

Priming/Backwashing

Flushing the SlideMaker

LH SlideStainer

Basket Holders

Basket Tray (s)

Bath Liquid Level Sensor

Bath Tray

Bath Tray Overflow Sensor

Drain and Fill Tubes

Dryer and Adjacent Plate

Holding Position Tray

Plastic Baths

Tubing Tray

Flushing Stain Baths and Tubing


55 of 59

LH 700 Series


See Cleaning Overview in HELP to determine if certain cleaning procedures are necessary. In HELP, go to the Index tab. Type in the word Cleaning. Select overview-diluter.

Other cleaning procedures found here include the following:

COMPONENT PROCEDURE TITLE

air filter Cleaning and Replacing Air Filters

aspiration probe wipe Cleaning Manual Aspiration Probe Wipe

BSV drip plate Cleaning Blood Sampling Valve Drip Plate

BSV outside Cleaning Blood Sampling Valve (Outside Sections)

clogged flow cell Clearing Clogged Flow Cell

needle assembly Clean Needle Assembly Area

rocker bed Rocker Bed

stripper plate Stripper Plate

Shear Valve (from search tab) Cleaning the Shear Valve


56 of 59

LH SlideMaker


To access the following cleaning procedures in HELP:

select Contents>LH SlideMaker>Cleaning Procedures


Air filter Cleaning and Replacing Air Filter (SlideMaker)

Blood Spills and Broken Slides Cleaning Blood Spills and Broken Slides

Catch Tray Cleaning a Catch Tray

Dispense Probe and Rinse Cup Cleaning the Dispense Probe and Rinse Cup

Priming/Backwashing Priming/Backwashing SlideMaker

Flushing the SlideMaker Flushing the SlideMaker (Bleach System)


57 of 59

LH SlideStainer As Needed Cleaning Procedures

To access the following cleaning procedures in HELP:

select Contents>LH SlideStainer>Cleaning Procedures.


Basket Holders Basket Holders

Basket Tray Basket Tray

Bath Liquid Level Sensor Bath Liquid Level Sensor

Bath Tray Bath Tray

Bath Tray Overflow Sensor Bath Tray Overflow Sensor

Drain and Fill Tubes Drain and Fill Tubes

Dryer and Adjacent Plate Dryer and Adjacent Plate

Holding Position Tray Holding Position Tray

Plastic Baths Plastic Baths

Tubing Tray Tubing Tray

Flushing Stain Baths and Tubing Flushing Stain Baths and Tubing

PN

A86683A

A (

Octo

ber

2009)

Mia

mi E

ducation C

ente

r 58

of

59

Op

tim

izin

g S

tain

Pro

toc

ols

Ove

rall

Sta

in Q

uali

ty

Beckm

an C

oulter

has d

evelo

ped

BC

I T

ruC

olo

r™ r

eage

nts

and

defa

ult p

roto

cols

fo

r use o

n t

he L

H S

lide

Sta

ine

r. T

he B

eckm

an C

oulter

defa

ult

pro

tocols

are

to

a p

redefin

ed c

olo

ration s

pecific

atio

n (

See ta

ble

s o

n n

ext p

age).

Sin

ce c

olo

ration is s

ubje

ctive,

optim

iza

tion o

f th

e p

roto

cols

and/o

r re

age

nts

ma

y b

e n

ecessary

to m

eet

your

ind

ivid

ua

l co

lora

tion c

rite

ria. E

nsure

that

both

the

meth

anol an

d w

ate

r yo

u u

se m

eet th

e s

pe

cific

atio

ns

deta

iled in y

our

sys

tem

’s o

nlin

e H

elp

.

Typ

ica

l ti

me

ad

jus

tme

nts

in

cre

me

nts

•

Sta

in –

30 s

econds

• B

uff

er

– 1

min

ute

Co

lora

tio

n (

red

/ b

lue)

is p

rim

ari

ly c

on

tro

lle

d b

y t

he

pH

of

the

bu

ffe

r.

Sta

in i

nte

ns

ity a

nd

dif

fere

nti

al

sta

inin

g c

ha

rac

teri

sti

cs c

an

are

co

ntr

olle

d b

y:

• th

e len

gth

of

sta

inin

g t

imes (

long

er

= m

ore

inte

nse)

• decre

asin

g th

e b

uff

er

str

ength

•

concentr

ation

of

sta

in in b

uff

er

(more

sta

in =

dark

er,

more

in

tense)

Sym

pto

m

Cau

se

Ac

tio

n

Buff

er

too a

cid

ic

a)

Adju

st

with a

lkalin

e s

olu

tion

until yo

u a

ch

ieve t

he d

esir

ed c

olo

r.

b)

Try

a b

uff

er

with

a h

igher

pH

.

Exce

ssiv

e w

ash

ing

a)

Short

en r

inse t

ime (

Bath

5)

until you

achie

ve th

e d

esire

d c

olo

r.

b)

Adju

st fr

equency o

f B

ath

5 d

rain

an

d f

ill u

ntil yo

u a

ch

ieve t

he d

esire

d c

olo

r.

All

rea

gents

too o

ld

Dra

in a

nd

fill

all

bath

s.

Sta

in t

ime too s

hort

P

rolo

ng s

tain

ing tim

e u

ntil you a

ch

ieve t

he d

esire

d c

olo

r.

To

o r

ed

Buff

er

tim

e too lon

g

Short

en b

uff

er

tim

e u

ntil yo

u a

chie

ve th

e d

esired c

olo

r.

Buff

er

too a

lkalin

e

a)

Adju

st

with a

cid

so

lutio

n u

ntil you

achie

ve th

e d

esire

d c

olo

r.

b)

Try

a b

uff

er

with a

lo

wer

pH

.

Insuff

icie

nt

wash

ing

Leng

the

n r

inse

tim

e u

ntil yo

u a

chie

ve th

e d

esired c

olo

r.

Sta

in t

ime too long

Decre

ase s

tain

tim

e u

ntil you a

ch

ieve t

he d

esire

d c

olo

r.

To

o b

lue

Buff

er

tim

e too s

hort

Leng

the

n b

uff

er

tim

e u

ntil you a

ch

ieve t

he d

esire

d c

olo

r.

Sta

in t

ime too long

Short

en s

tain

tim

e u

ntil you

achie

ve

the

desir

ed c

olo

r.

To

o d

ark

Buff

er

tim

e too lon

g

Short

en b

uff

er

tim

e u

ntil yo

u a

chie

ve th

e d

esired c

olo

r.

Rin

se t

ime too long

Decre

ase r

inse

tim

e u

ntil you a

ch

ieve t

he d

esire

d c

olo

r.

Poor

qua

lity r

inse w

ate

r a)

Chan

ge w

ate

r.

b)

Verify

wate

r m

eets

specific

ations o

f T

ype II

NC

CL

S w

ate

r

Sta

in t

ime too s

hort

In

cre

ase s

tain

tim

e u

ntil yo

u a

chie

ve th

e d

esired c

olo

r

To

o lig

ht

Buff

er

tim

e too s

hort

In

cre

ase b

uff

er

tim

e u

ntil you a

ch

ieve t

he d

esire

d c

olo

r

PN

A86683A

A (

Octo

ber

2009)

Mia

mi E

ducation C

ente

r 59

of

59

Buff

er

too c

oncentr

ate

d

Dilu

te b

uff

er

solu

tio

n w

ith d

eio

niz

ed

or

dis

tille

d w

ate

r u

ntil yo

u a

chie

ve

the

desired

colo

r

Sym

pto

m

Cau

se

Ac

tio

n

RB

C c

en

ters

a

pp

ea

r p

un

ch

ed

o

ut

(co

okie

cu

tter

eff

ect

in

cen

ter

of

cells)

Wate

r has c

onta

min

ate

d

the m

eth

ano

l a)

Dra

in/F

ill t

he m

eth

an

ol b

ath

. b)

Dip

em

pty

basket in

meth

ano

l pri

or

to r

e-e

ntr

y into

syste

m.

Sta

in t

oo o

ld

Dra

in/f

ill S

tain

and S

tain

/Bu

ffer

bath

s.

Backg

rou

nd

sta

inin

g (

ov

era

ll

slid

e s

ho

ws

co

lor)

Exce

ss s

tain

left

on

slid

e

Add q

uic

k r

inse s

tep to p

roto

col (p

rim

arily

for

WG

and M

GG

). S

ee

he

lp t

op

ic Q

uic

k R

inse

Meth

od.

Sta

in o

r S

tain

/buff

er

too

old

Dra

in/F

ill a

ll bath

s.

Rin

se (

Bath

5)

dirty

a)

Dra

in/F

ill B

ath

5.

b)

Incre

ase f

reque

ncy o

f B

ath

5 d

rain

/fill

. c)

Add Q

uic

k R

inse s

tep t

o p

roto

col (p

rim

arily

for

WG

or

MG

G).

See h

elp

top

ic Q

uic

k R

inse

Meth

od.

Pre

cip

itate

(c

lum

ps o

r fi

ne

part

icle

s o

n

wh

ole

slid

e)

Sta

in b

ath

s d

irty

(S

tain

and S

tain

/Buff

er)

P

erf

orm

the B

ath

Flu

shin

g P

rocedure

.

Ov

era

ll p

ale

blu

e

sta

inin

g

Blo

od s

mear

more

than

2 h

ours

old

prior

to f

ixin

g

Make n

ew

blo

od s

mear.

Th

e t

ab

les b

elo

w r

ep

res

en

t th

e s

tain

ch

ara

cte

risti

cs a

gain

st

wh

ich

th

e B

eck

man

Co

ult

er

Tru

Co

lor™

reag

en

ts a

nd

BC

I d

efa

ult

pro

toco

ls w

ere

ev

alu

ate

d.

R

efe

ren

ce

: Win

trobe’s

Clin

ical H

em

ato

logy, 9

th e

ditio

n, V

ol. 1

, page 2

3.

NC

CLS

Docum

ent H

20 A

, V

olu

me 1

2, N

um

ber

1. R

efe

rence L

eukocyte

Diffe

rential C

ount (P

roport

ional) a

nd E

valu

ation o

f In

str

um

enta

l m

eth

ods.

Colo

r A

tlas a

nd Instr

uction M

anual of P

eriphera

l C

ell

Morp

holo

gy, O

’Connor,

Barb

ara

H., 1

984, page 1

6 –

18.C

opie

d f

rom

: P

N 4

276448D

(N

ovem

ber

2003)

Co

lori

ng

of

Ro

man

ow

sk

y S

tain

s

Str

uctu

re

Cell

Co

lor

Nucle

i A

ll P

urp

le

Ery

thro

cyte

P

ink

Lym

phocyte

B

lue

Monocyte

G

ray b

lue

Neutr

op

hil

Pin

k

Meta

myelo

cyte

P

ink

Mye

locyte

P

ink

Cyto

pla

sm

Pro

mye

locyte

B

lue

Gra

nule

s

Basop

hil

Purp

le,

bla

ck

E

osin

op

hil

Ora

nge

N

eutr

op

hil

Purp

le

P

late

let

Purp

le

Hu

man

Blo

od

Ce

ll In

clu

sio

ns

Inclu

sio

n

Desc

rip

tio

n

Cell

Co

lor

Auer

rods

Spic

ule

-lik

e f

orm

ation

M

ye

lobla

st

Pin

k

Pap

pen

he

imer

bod

ies

Sm

all

spots

R

ed b

lood c

ells

P

urp

le

Mala

ria p

ara

sites

Inclu

sio

ns

Red b

lood c

ells

P

urp

le/b

lue

Ho

we

ll-Jolly

bo

die

s

Med

ium

siz

ed s

pot

Red b

lood c

ells

P

urp

le

Cabot

rin

gs

Thre

adlik

e f

ilam

ents

R

ed b

lood c

ells

P

urp

le

Ba

so

phili

c s

tipp

ling

Sm

all

gra

nule

s

Red b

lood c

ells

P

urp

le

Toxic

Gra

nula

tion

C

oars

e G

ranu

les

WB

C

Dark

Purp

le

Dohle

bodie

s

Spots

W

BC

S

ky-

Blu

e

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