Beckman Coulter Coulter LH 700 Series Service manual

COULTER LH 750 System
Reference
PN 4277248DD (January 2013)
Beckman Coulter, Inc.
250 S. Kraemer Blvd.
Brea, CA 92821 U.S.A.
*SYSTEM RUN*
04/30/96
12:30:18
STATUS:R EADY
ALERT:
MODE:C/D/R
BLOOD DETECT: ON
TEST MODE
ASPIRATIONS/TUBE01
MAIN MENU
VIII
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VI
V
IV
III
II
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POWERON
POWEROFF
STARTUP
SHUTDOWN
ALARM
RESET
READY
PRIME
APERT DRAIN RINSE CLEAR
APERT
CE
7
4
1F 2
5
8
0
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ID
STOP PREMIXSTART
CONT
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NEC MultiSync LCD 1700m+

WARNINGS AND PRECAUTIONS
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 IT IS NOT LIMITED TO,
PROTECTIVE EYEWEAR, GLOVES, AND SUITABLE LABORATORY ATTIRE WHEN OPERATING OR MAINTAINING THIS OR
ANY OTHER AUTOMATED LABORATORY ANALYZER.
HAZARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS
WARNINGS, CAUTIONS, and IMPORTANTS alert you as follows:
WARNING - Can cause injury.
CAUTION - Can cause damage to the instrument.
IMPORTANT - Can cause misleading results.
WARNING Risk of operator injury if:
r All doors, covers and panels are not closed and secured in place prior to and during instrument operation.
r The integrity of safety interlocks and sensors is compromised.
r Instrument alarms and error messages are not acknowledged and acted upon.
r You contact moving parts.
r You mishandle broken parts.
r Doors, covers and panels are not opened, closed, removed and/or replaced with care.
r Improper tools are used for troubleshooting.
To avoid injury:
r Keep doors, covers and panels closed and secured in place while the instrument is in use.
r Take full advantage of the safety features of the instrument. Do not defeat safety interlocks and sensors.
r Acknowledge and act upon instrument alarms and error messages.
r Keep away from moving parts.
r Report any broken parts to your Beckman Coulter Representative.
r Open/remove and close/replace doors, covers and panels with care.
r Use the proper tools when troubleshooting.
CAUTION System integrity might be compromised and operational failures might occur if:
r This equipment is used in a manner other than specified. Operate the instrument as instructed in the Product Manuals.
r You introduce software that is not authorized by Beckman Coulter into your computer. Only operate your system’s
computer with software authorized by Beckman Coulter.
r You install software that is not an original copyrighted version. Only use software that is an original copyrighted
version to prevent virus contamination.
IMPORTANT If you purchased this product from anyone other than Beckman Coulter or an authorized Beckman Coulter
distributor, and, if 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.

iii
PN 4277248DD
REVISION STATUS
Issue A, 10/01
Software version 1A. Converted from Help Version 1A.012431.
Issue B, Complete Revision, 5/02
Software version 2A. Converted from Help Version 2A.021501.
Issue C, 10/03
Software version 2B. Converted from Help Version 2B.031971.
Changes were made to:
r Change the company name from Coulter Corporation to Beckman Coulter Inc.
r Change LH 750 to LH 700 Series.
r Change all variations of bar-code to be consistent.
r Change 5C-ES to 5C Series.
Issue D, 04/07
Software version 2D. Converted from Help Version 2D.071161. The name of the manual
was changed from COULTER LH 700 Series System Reference to COULTER LH 750 System
Reference.
Issue DA, 08/09
Software version 2D1. Converted from Help Version 2D1.091732.
Issue DB, 11/10
Software Version 2D1.
Updates were made to the company corporate address.
Issue DC, 12/10
Software Version 2D1.
Changes were made to:
r BAR CODES AND THE LH 700 Series
r Checksum Algorithm
r NW-7
r Code 128/USS 128
r BAR-CODE LABEL SPECIFICATIONS
r Acceptable Bar Codes
r Codabar
r NW7 Decoding
Note: Changes that are part of the most recent revision are indicated in text by a bar in the
margin of the amended page.
This document applies to the latest software listed and higher versions. When a subsequent software version
changes the information in this document, a new issue will be released to the Beckman Coulter website. For
labeling updates, go to www.beckmancoulter.com and download the most recent manual or system help for
your instrument.

PN 4277248DD
iv
REVISION STATUS
Issue DD, 01/13
Software Version 2D3. Manual derived from Online Help version 2D3.
Changes were made to:
r SPECIAL REQUIREMENTS: HARDWARE
r Space and Accessibility
r Pneumatic/Hydraulic Tubing Connections
r Changing 9-pin and 25-pin connectors
r Added reference to the Hematology Tube List information
Note: Changes that are part of the most recent revision are indicated in text by a bar in the
margin of the amended page.
This document applies to the latest software listed and higher versions. When a subsequent software version
changes the information in this document, a new issue will be released to the Beckman Coulter website. For
labeling updates, go to www.beckmancoulter.com and download the most recent manual or system help for
your instrument.

PN 4277248DD v
CONTENTS
WARNINGS AND PRECAUTIONS, ii
REVISION STATUS, iii
CONTENTS, v
INTRODUCTION, xiii
HOW TO USE YOUR COULTER LH 750 SERIES SYSTEM HARD-COPY MANUALS, xiii
ABOUT THIS MANUAL, xiv
ONLINE HELP SYSTEM, xiv
CONVENTIONS, xiv
1 USE AND FUNCTION, 1-1
1.1 INTENDED USE, 1-1
1.2 INDICATIONS FOR USE, 1-1
Parameters, 1-2
1.3 QUALITY CONTROL (QC), 1-3
1.4 METHOD HISTORY, 1-3
Development, 1-3
Corrected WBC Counts, 1-4
Hemoglobinometry, 1-4
Differential Measurement, 1-4
Volume Analysis, 1-4
Conductivity Analysis, 1-5
Light Scatter Analysis, 1-5
Reticulocyte (Retic) Analysis, 1-5
NRBC Enumeration, 1-6
COULTER IntelliKinetics Application, 1-7
XB Analysis, 1-7
XM Analysis, 1-7
Extended QC, 1-7
1.5 SYSTEM COMPONENTS, 1-8
Power Supply, 1-8
Diluter, 1-8
Analyzer, 1-8
LH Workstation, 1-8
Handheld Scanner, 1-8
1.6 HARDWARE OPTIONS, 1-9
Graphic/Laser Printer, 1-9
LH SlideMaker, 1-9
LH SlideStainer, 1-9

PN 4277248DD
vi
CONTENTS
1.7 CONTROLS AND CALIBRATOR, 1-9
Controls, 1-9
Calibrator, 1-9
1.8 REAGENTS, 1-9
Diluent, 1-10
CBC Lytic Reagent, 1-10
LH Series PAK Reagent System, 1-10
The LH Series RETIC PAK Reagent Kit, 1-10
Cleaners, 1-11
1.9 MATERIAL SAFETY DATA SHEETS (MSDS), 1-11
2 INSTALLATION, 2-1
2.1 GENERAL, 2-1
2.2 SPECIAL REQUIREMENTS: HARDWARE, 2-1
Space and Accessibility, 2-1
Electrical Input, 2-2
Ambient Temperature and Humidity, 2-3
Air Conditioning, 2-3
Ventilation, 2-3
Drainage, 2-3
2.3 INTERUNIT CONNECTIONS, 2-4
Power and Signal Cables, 2-4
Changing 9-pin and 25-pin connectors, 2-4
Pneumatic/Hydraulic Tubing Connections, 2-5
3 OPERATION PRINCIPLES, 3-1
3.1 COULTER METHOD, 3-1
CBC Analysis, 3-1
Differential Analysis, 3-1
Effect of Reagents, 3-2
Reticulocyte Analysis, 3-2
3.2 AUTOMATIC ASPIRATION MODE, 3-2
Loading Specimens, 3-2
Transporting Cassettes, 3-2
Aspiration, 3-4
Delivery, 3-4
CBC, 3-4
Differential (Diff), 3-4
Reticulocyte (Retic), 3-5
CBC Sensing System, 3-6
CBC Analysis in the Baths, 3-6
Diff and Retic Multiparameter Sensing System, 3-6
Backwash and Rinse, 3-8
3.3 MANUAL ASPIRATION MODE, 3-8

PN 4277248DD vii
CONTENTS
3.4 COUNTING AND SIZING, 3-9
Red and White Blood Cell Counting, 3-9
Coincidence Correction, 3-10
Voting, 3-10
Pulse Editing, 3-10
Sweep Flow, 3-10
RBC Size Distribution, 3-11
Platelet Method, 3-11
3.5 MEASUREMENT OF HEMOGLOBIN CONCENTRATION, 3-12
3.6 DATAPLOT DEVELOPMENT, 3-13
Two-Dimensional (2D) DataPlots, 3-13
Three-Dimensional (3D) DataPlots, 3-13
3.7 PARAMETERS AND THEIR DERIVATION, 3-14
Summary of Parameter Derivations, 3-14
Measured Directly, 3-14
Derived From RBC or Plt Histogram, 3-14
Derived From WBC Histogram, 3-15
Calculated, 3-15
White Blood Cell (WBC) Count, 3-15
Red Blood Cell (RBC), 3-15
Hemoglobin (Hgb) Concentration, 3-15
Mean Corpuscular Volume (MCV), 3-16
Hematocrit (Hct), 3-16
Mean Corpuscular Hemoglobin (MCH), 3-16
Mean Corpuscular Hemoglobin Concentration (MCHC), 3-16
Red Cell Distribution Width (RDW), 3-16
Platelet (Plt) Count, 3-16
Mean Platelet Volume (MPV), 3-16
NRBC % (Nucleated Red Blood Cells), 3-16
NRBC # (Nucleated Red Blood Cells), 3-17
Differential (Diff), 3-17
Diff Percentages (DIFF%), 3-17
Diff Absolute Numbers (DIFF#), 3-17
Reticulocyte (Retic) Parameters, 3-18
Retic Percent (RET%), 3-18
Retic Absolute Number (RET#), 3-18
Immature reticulocyte fraction (IRF), 3-18
Mean reticulocyte volume (MRV), 3-18
3.8 XB ANALYSIS, 3-19
Adjusting Initial XB Target Values, 3-20
XM Analysis, 3-21
Extended QC, 3-21

PN 4277248DD
viii
CONTENTS
4 SPECIFICATIONS/CHARACTERISTICS, 4-1
4.1 PHYSICAL SPECIFICATIONS, 4-1
Dimensions, 4-1
Power, 4-1
Input, 4-1
Consumption, 4-1
4.2 PERFORMANCE SPECIFICATIONS--LH 700 Series, 4-1
Precision, 4-1
Within-Run Precision, 4-1
Accuracy, 4-2
Accuracy Qualification, 4-2
Accuracy, CBC, 4-2
Accuracy, WBC Differential, 4-3
Accuracy, Reticulocyte, 4-3
Linearity, 4-3
Background, 4-4
Carryover, 4-4
Operating and Reportable Ranges, 4-4
Mode-to-Mode Comparison, 4-5
4.3 PERFORMANCE CHARACTERISTICS, 4-5
Sample Stability, 4-6
WBC Differential Flagging Stability, 4-8
NRBC Paired Sample Imprecision, 4-9
MCV Accuracy Characteristics, 4-9
NRBC Accuracy Characteristics, 4-10
Platelet Accuracy Characteristics, 4-10
Reference Ranges, 4-10
Temperature, 4-11
Humidity, 4-11
Storing Specimens, 4-11
Venipuncture Specimens, 4-11
Sample Type, 4-12
Recommended Anticoagulant, 4-12
Sampling Modes, 4-12
Aspiration, 4-12
Test/Cycle, 4-12
Throughput, Automatic Mode, 4-12
Sample Volume Aspirated, 4-12
Waste, 4-12
Pneumatic Supplies (Internally Regulated), 4-13
Calibration Stability, 4-13
LH Workstation Storage, 4-13
Known Interfering Substances, 4-13
CBC, 4-13
NRBC, 4-14
Differential, 4-14
Reticulocytes, 4-14

PN 4277248DD ix
CONTENTS
4.4 BAR-CODE SYMBOLOGY OVERVIEW, 4-14
Character Set, 4-15
Symbology Type, 4-15
Fixed or Variable Length, 4-15
Self-Checking, 4-15
Start Code, Stop Code, 4-15
Check Character, Checksum Algorithm, 4-15
Quiet Zone, Quiet Area, 4-15
4.5 BAR CODES AND THE LH 700 Series, 4-16
Checksum Algorithm, 4-16
Bar-Code Symbologies Supported by the LH 700 Series, 4-16
Interleaved 2-of-5, 4-16
Code 39 (Also called 3-of-9 Code), 4-16
Codabar, 4-17
NW-7, 4-17
Code 128/USS 128, 4-17
Bar-Code Tips, 4-17
4.6 BAR-CODE LABEL SPECIFICATIONS, 4-18
General, 4-18
Optical Characteristics at 880 nm ±10% and 633 nm ±10%, 4-18
Printing Method, 4-18
Label Thickness, 4-18
NE/WE Ratio, 4-18
Label Dimensions and Data, 4-19
Acceptable Bar Codes, 4-20
Extended Digit Bar Code, 4-20
Checksum Algorithm, 4-21
Interleaved 2-of-5, 4-21
Codabar, 4-22
NW7 Decoding, 4-23
Code 39 Bar Code, 4-25
Code 128, 4-26
5 HAZARDS, 5-1
5.1 LASER SAFETY, 5-1
5.2 RADIATION HAZARDS, 5-1
5.3 LASER WARNING LABELS, 5-1
5.4 BAR-CODE READER, 5-3
5.5 HANDHELD SCANNER, 5-5
5.6 DISPOSAL OF ELECTRICAL INSTRUMENTATION, 5-5

PN 4277248DD
x
CONTENTS
REFERENCES, REFERENCES-1
GLOSSARY, GLOSSARY-1
INDEX, INDEX-1
BECKMAN COULTER, INC. CUSTOMER END USER LICENSE AGREEMENT, 1
TRADEMARKS, 3

PN 4277248DD xi
CONTENTS
ILLUSTRATIONS
1.1 COULTER LH 700 Series, 1-1
1.2 Corrected WBC, 1-4
1.3 Illustration of the ten light scatter regions, 1-5
1.4 NRBC signature position on Differential Dataplot, 1-6
1.5 NRBC location on WBC histogram, 1-6
2.1 Space for tower-type computers, 2-1
2.2 Space for Small Form Factor (SFF)-type computers, 2-2
2.3 Rear of instrument and accessories with DB-25 pin for tower-type computers, 2-4
2.4 Rear of instrument with DB 9-Pin Connector, 2-5
2.5 Pneumatic/Hydraulic Connections, 2-6
3.1 Coulter Method of Counting and Sizing, 3-1
3.2 Transport System, 3-3
3.3 Triple Transducer Module with Protective Housing, 3-7
3.4 Triple Transducer Module with Protective Housing Cut Away, 3-8
3.5 Sweep Flow, 3-11
4.1 Bar-Code Label Specifications, 4-19
5.1 Laser Warning Label, Protective Housing Cut Away, 5-2
5.2 Laser Warning Label Locations, Protective Housing On, 5-3
5.3 Bar-Code Reader Laser Warning Label Location, Protective Housing Cut
Away, 5-3
5.4 Bar-Code Reader Laser Warning Label Location, Protective Housing On, 5-4
5.5 Handheld Scanner, 5-5

PN 4277248DD
xii
CONTENTS
TABLES
2.1 Description of Pneumatic/Hydraulic Connections, 2-6
3.1 Effect of Directly-Measured Parameters on the Red Cell Indices, 3-21
4.1 Within-Run Precision, 4-2
4.2 Accuracy, CBC, 4-2
4.3 Accuracy Tolerance Limits, WBC Differential, 4-3
4.4 Accuracy, Reticulocyte, using a predicate hematology analyzer, 4-3
4.5 Linearity Limits, 4-4
4.6 Operating and Reportable Ranges, 4-5
4.7 CBC Sample Stability, Room Temperature‡, 4-6
4.8 CBC Sample Stability, Cold Temperature‡, 4-6
4.9 DIFF% Sample Stability, Room Temperature‡, 4-6
4.10 DIFF% Sample Stability, Cold Temperature‡, 4-6
4.13 RETIC Sample Stability, Room Temperature‡, 4-7
4.11 DIFF# Sample Stability, Room Temperature‡, 4-7
4.12 DIFF# Sample Stability, Cold Temperature‡, 4-7
4.15 NRBC Sample Stability, Room Temperature‡, 4-8
4.16 NRBC Sample Stability, Cold Temperature‡, 4-8
4.14 RETIC Sample Stability, Cold Temperature‡, 4-8
4.17 Differential Suspect Flagging at Room Temperature, 4-9
4.18 Differential Suspect Flagging at Cold Temperature, 4-9
4.19 NRBC Imprecision Characteristics,Paired Sample Analysis - Replicate 1 vs.
Replicate 2, 4-9
4.20 MCV Performance Characteristics, 4-9
4.21 NRBC Accuracy Characteristics, Compared Samples, 4-10
4.22 Plt Accuracy Characteristics vs ICSH/ISLH Platelet Method, 4-10
4.23 Normal Population Study, 4-10
4.24 Bar-Code Label Specifications, 4-19
4.25 Code-Related Specifications, 4-21
4.26 Codabar Character Value Table, 4-22
4.27 NW 7 Character Value Table, 4-23
4.28 Code 39 Character Value Table:, 4-25

PN 4277248DD xiii
INTRODUCTION
This introductory section contains the following topics:
r How to use your COULTERLH 750 System hard-copy manuals
r About this manual
r Online Help System
r Conventions
HOW TO USE YOUR COULTER LH 750 SERIES SYSTEM HARD-COPY MANUALS
Use the Getting Started booklet to see an overview of the system hardware and software. This
document comes with your LH 750 System.
Use the Reference manual for in-depth information about what the instrument does, the
methods it uses, its specifications, and information on installation, safety and software
options. The Reference manual for the LH 750 System is included in the online Help system;
it is available in hard copy by request.
Use the Special Procedures and Troubleshooting manual to run calibration; to clean, replace
or adjust a component on the instrument; and for troubleshooting the instrument. This
document is made up of procedures from the online Help system; it is available in hard copy
by request.
Use the Operator’s Guide for the day-to-day operation of your instrument. This document is
made up of procedures from the online Help system; it includes Startup, running controls and
samples, reviewing data, Shutdown, and the software on the Analyzer and the Workstation.
This document is available in hard copy by request.
Use the SlideMaker Operator’s Guide for in-depth information about what the SlideMaker
does, the methods it uses, its specifications, and information on installation, safety and
software, as well as day-to-day operating and troubleshooting your SlideMaker. This
document is made up of procedures from the online Help system; it is available in hard copy
by request.
Use the SlideStainer Operator’s Guide for the day-to-day operating and troubleshooting of
your SlideStainer. This document is made up of procedures from the online Help system; it
includes in-depth information about what the SlideStainer does, the methods it uses, its
specifications, and information on installation, safety and software. This document is
available in hard copy by request.
Use the Master Index to easily locate a subject in your hard-copy Reference manual,
Operator’s Guide or Special Procedures and Troubleshooting manual. The Master Index
comes with the hard copy of both the Operator’s Guide and the Special Procedures and
Troubleshooting manual.
Use the Host Transmission Specification to find the information needed to program the
transmission interface between the LH 750 System and your laboratory’s host computer. This
document is available in hard copy by request.
See the Documentation page on the back cover of this manual for the contents of each
manual. It can help you to determine quickly in which manual the information you need is
located.

PN 4277248DD
xiv
INTRODUCTION
ABOUT THIS MANUAL
ABOUT THIS MANUAL
Your LH 750 System Reference Guide is a source of information for the day-to-day operation
of your instrument. This information is organized as follows:
s Chapter 1, Use and Function
Contains the intended use of the instrument, a brief history of the methods used, the
reagents, calibrators and controls used, and a short description of the major components.
s Chapter 2, Installation
Contains the instrument requirements, and the diagrams of the reagent/pneumatic
tubing connections and the interunit cable connections.
s Chapter 3, Operation Principles
Contains descriptions of the Coulter Method, the normal sample flow through the
instrument, how counting and sizing are accomplished, and what the DataPlots show.
s Chapter 4, Specifications/Characteristics
Details the instrument and performance specifications, the performance characteristics,
the interferring substances, and the bar-code label specifications.
s Chapter 5, Hazards
Describes laser safety precautions and the location of the laser-related labels.
s References
s Glossary
s Index, hard copy only
ONLINE HELP SYSTEM
The LH Workstation has a comprehensive Online Help System, which includes reference
information, all operating, maintenance and troubleshooting procedures. On the LH
Workstation, select to access Help. Select to access the tutorials.
CONVENTIONS
This document uses the following conventions:
indicates a key on the Numeric keypad.
indicates a key on the LH Workstation keyboard.
is the icon for Patient results on the LH Workstation.
is the icon for the Printer on the LH Workstation.

PN 4277248DD 1-1
1
1USE AND FUNCTION
1.1 INTENDED USE
The COULTER LH 750 Hematology Analyzer is a quantitative, automated hematology
analyzer and leukocyte differential counter For In Vitro Diagnostic Use in clinical
laboratories. The COULTER LH 750 Hematology Analyzer provides automated Reticulocyte
analysis and enumeration of nucleated red blood cells (NRBCs) as well as an automated
method for enumeration of RBCs and WBCs in body fluids.
1.2 INDICATIONS FOR USE
The purpose of the LH 700 Series (Figure 1.1) is to separate the normal patient, with all
normal system-generated parameters, from the patient who needs additional studies of any of
these parameters. These studies might include further measurements of cell size and platelet
distribution, manual WBC differential or any other definitive test that helps diagnose the
patient’s condition.
Figure 1.1 COULTER LH 700 Series
N E C
M u l t i S y n c L C D 1 7 0 0 m +
G
EN•
S
* S Y S T E M R U N *
04/30/96
1 2 : 3 0 : 1 8
S T A T U S : R E A D Y
A L E R T :
M O D E : C / D / R
B L O O D D E T E C T : O N
T E S T M O D E
A S P I R A T I O N S / T U B E 0 1
M A I N M E N U
VIII
VII
VI
V
IV
III
II
I

PN 4277248DD
1-2
USE AND FUNCTION
INDICATIONS FOR USE
Parameters
The system determines these hematologic parameters of whole-blood specimens:
*For Research Use Only. Not For Use In Diagnostic Procedures.
Unless otherwise stated, all parameter results are shown in a US unit format throughout the
manuals.
WBC White Blood Cell or leukocyte count
RBC Red Blood Cell or erythrocyte count
Hgb Hemoglobin concentration
Hct Hematocrit (relative volume of erythrocytes)
MCV Mean Corpuscular (erythrocyte) Volume
MCH Mean Corpuscular (erythrocyte) Hemoglobin
MCHC Mean Corpuscular (erythrocyte) Hemoglobin Concentration
RDW Red Cell (erythrocyte volume) Distribution Width
Plt Platelet or thrombocyte count
MPV Mean Platelet (thrombocyte) Volume
LY% Lymphocyte percent
MO% Monocyte percent
NE% Neutrophil percent
EO% Eosinophil percent
BA% Basophil percent
LY# Lymphocyte number
MO# Monocyte number
NE# Neutrophil number
EO# Eosinophil number
BA# Basophil number
NRBC% Nucleated Red Blood Cell percent
NRBC# Nucleated Red Blood Cell number
RET% Reticulocyte percent
RET# Reticulocyte number
*HLR% High Light scatter Reticulocytes %
*HLR# High Light scatter Reticulocytes #
IRF Immature Reticulocyte Fraction
MRV Mean Reticulocyte Volume
*MSCV Mean Sphered Cell Volume
*Pct Plateletcrit
*PDW Platelet Distribution Width

PN 4277248DD 1-3
USE AND FUNCTION
QUALITY CONTROL (QC) 1
1.3 QUALITY CONTROL (QC)
Your laboratory can use these QC techniques with the LH 700 Series:
r Daily instrument checks
r Commercial or Patient Controls
r Delta checks
r XB Analysis
r Extended QC
r XM Analysis
r Interlaboratory Quality Assurance Program (IQAP)
Quality Assurance includes routine maintenance and service in conjunction with the use of
controls and calibrators. The combination of these methods provides the assurance of
complete quality control and should be applied separately or in combination, in accordance
with your laboratory, state and federal protocols.
1.4 METHOD HISTORY
Development
W.H. Coulter (1956) describes the Coulter Principle:1
A suspension of blood cells is passed thru [sic] a small orifice simultaneously with an
electric current. The individual blood cells passing thru the orifice introduce an
impedance change in the orifice determined by the size of the cell. The system counts the
individual cells and provides cell size distribution. The number of cells counted per
sample is approximately 100 times greater than the usual microscope count to reduce the
statistical error by a factor of approximately 10 times.
This substantial improvement in precision over previous methods helped to establish the
erythrocyte count as a sensitive index of erythropoietic dyscrasia, particularly when
considered together with Hct and Hgb measurements.2
The COULTER COUNTER Model S analyzer was the first instrument that automated
simultaneous multiparameter measurements on blood. Brittin et al., Gottmann, and Hamilton
and Davidson, reviewed the performance and clinical value of the Model S.3, 4, 5
Refinements of the COULTER COUNTER analyzer to provide accurate size (volume)
distribution data led to a reawakening of interest in pathological erythrocyte size distribution,
first sparked by Price-Jones.6, 7
Among the advantages offered by the Coulter method of counting and sizing was the ability
to derive an accurate Hct measurement by summing the electronic volume of erythrocytes.
England et al. speculated that electronic Hct measurements did not contain the trapped
plasma error of centrifugal Hct measurements.8
Bull et al. described the use of a COULTER COUNTER analyzer for counting thrombocytes.9
This method, useful as it was, depended on preparing thrombocyte-rich plasma to avoid
counting erythrocytes as thrombocytes. Mundschenk et al. and Schulz and Thom discussed
the possibility of counting thrombocytes in the presence of erythrocytes and classifying them

PN 4277248DD
1-4
USE AND FUNCTION
METHOD HISTORY
by size.10, 11 Electronic refinements in the Model S-PLUS enhanced the accuracy of the
hydrodynamic method. Von Behrens and Paulus have also cited the feasibility of counting
thrombocytes by the Coulter method.12, 13
Corrected WBC Counts
White Blood Cell count results from the CBC analysis. The WBC count may be adjusted for
interfering substances when appropriate. If there is a population of cells in the far left of the
WBC histogram, the number of cells is derived and the WBC count is corrected. No further
correction of WBC is required.
Figure 1.2 Corrected WBC
When WBC correction has occurred, the uncorrected WBC will appear in the printout
Comments field as UWBC = "value". The "Cellular Interference" suspect message is
displayed and the corrected WBC count is reported. The uncorrected WBC can be found on
the CBC data tab. When the separation between the WBC populations is poorly defined on
the histogram, WBC correction will be performed and the corrected WBC will have an R flag.
Hemoglobinometry
The lytic reagent used for the complete blood count (CBC) parameters prepares the blood so
the system can count leukocytes and measure the amount of hemoglobin. The lytic reagent
rapidly and simultaneously destroys the erythrocytes and converts a substantial proportion of
the hemoglobin to a stable pigment while it leaves leukocyte nuclei intact. The absorbance of
the pigment is directly proportional to the hemoglobin concentration of the sample.
The accuracy of this method equals that of the hemiglobincyanide method, the reference
method of choice for hemoglobinometry recommended by the International Committee for
Standardization in Hematology. 14
Differential Measurement
The COULTER VCS established WBC differential technology using three measurements:
individual cell volume, high-frequency conductivity and laser-light scatter.
The combination of low-frequency current, high-frequency current and light-scattering
technology provides abundant cell-by-cell information that is translated by the instrument
into conventional stained-film leukocyte categories.
Volume Analysis
Electronic leukocyte volume analysis, using low-frequency current, has been used since 1967.
15
It has been evaluated as a possible adjunct to the differential white cell count.
16,17,18,19

PN 4277248DD 1-5
USE AND FUNCTION
METHOD HISTORY 1
Conductivity Analysis
Cell walls act as conductors to high-frequency current. The current, while passing through
the cell walls and through each cell interior, detects differences in the insulating properties of
cell components. The current characterizes the nuclear and granular constituents and the
chemical composition of the cell interior. 20,21,22
Light Scatter Analysis
Coulter's experience in flow cytometry dates back decades to Fulwyler's pioneering use of
light scatter for cell analysis.23 Loken et al. and Jovin et al. discuss the relationship of particle
size and refractivity to the angle of light scattered from a laser beam.24,25
Reticulocyte (Retic) Analysis
Reticulocytes are immature, nonnucleated erythrocytes retaining a small network of
basophilic organelles, consisting of RNA and protoporphyrin. The enumeration of
reticulocytes provides a simple, effective means to determine red cell production and
regeneration.26,27,28,29
The most common means of measuring reticulocytes is to use supravital dyes, such as New
Methylene Blue or Brilliant Cresyl Blue. These dyes precipitate and aggregate the basophilic
substances within the reticulocyte, resulting in a granular, staining pattern easily seen with
light microscopy.30
Reticulocyte immaturity is related to cell volume and light scatter. Since more immature
reticulocytes are larger, contain more RNA and cause increased light scatter, the cell volume
and light scatter will increase with immaturity of the cell.
Figure 1.3 illustrates the IRF and MRV algorithms. This figure is a representation of the VCS
data that is shown on the two dimensional analyzer displays.
Figure 1.3 Illustration of the ten light scatter regions
The RET% is calculated as the ratio of reticulocytes to the total number of red cells. The
spectrum of light scatter intensity for the retic population is analyzed algorithmically. The
detected light scatter intensity of the retic population is divided into equal regions as shown
above. The IRF parameter is calculated as the ratio of the total number of retic events in the
outermost eight regions (3 to 10) to the total number of retics (regions 0 to 10 - region 0 is
not illustrated above). The MRV parameters is calcualted as the average volume of all
reticulocytes or the mean volume of all retic events.
1 1023456789
M a t u r e
r e d c e l l s R e t i c u l o c y t e s
H i g h l i g h t s c a t t e r r e t i c u l o c y t e s
P l a t e l e t s / d e r i s
M a t u r e
r e d c e l l s
R e t i c s W h i t e
c e l l s
L i g h t s c a t t e r
V o l u m e

PN 4277248DD
1-6
USE AND FUNCTION
METHOD HISTORY
NRBC Enumeration
The NRBC Enumeration is achieved through the combined use of impedance and VCS
technology and a proprietary algorithm.
The first step in NRBC enumeration is the identification of particles in the NRBC signature
position in the differential data plot. This information is generated from VCS analysis of the
cells.
Figure 1.4 NRBC signature position on Differential Dataplot
Once particles have been identified in this region, the LH 700 Series examines the far left
region of the WBC histogram for the presence of particles.
Figure 1.5 NRBC location on WBC histogram
If the VCS dataplot and the WBC histogram both indicate the presence of NRBCs, then the
combined information is further evaluated for special data patterns -- such as small
lymphocytes, giant platelets, and aging blood. If the combined information from the VCS
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