Dr.

ABDULRAHMAN AL MISHARI HOSPITAL

POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 1 oI 12
*. STATEMENT O+ PURPOSE:
1.1. To give inIormation about operating LH 750 Analyzer and Coulter Gen`s system in perIorming CBC
test in order to obtain qualiIied results. The coulter LH 750 Analyzer and Coulter Gen`s system is a
quantities automated hematology analyzer in vitro diagnostic use in clinical laboratories. The LH 750
Analyzer and Coulter Gen`s system provides automated complete blood count, leukocytes diIIerential
and a reticulocyte analysis.

,. DE+INITION:
2.1.CBC: Complete blood count.
2.2.T-e .en/& 0 123 Anal!4er and Co"lter .en/& &!&tem:
2.2.1.A multi-parameter automated Sample Hematology CBC analyzer
5. E6UIPMENT7MATERIAL7+ORMS:
3.1.Main unit
3.2.Sampler unit
3.3.CPU
3.4.Pneumatic unit
3.5.Reagents:
3.5.1.Dil"ent8 ISOTON III i& an i&otoni eletrol!te &ol"tion.
3.5.1.1.Dilutes the whole-blood sample.
3.5.1.2.Stabilizes cells membranes Ior accurate counting and sizing.
3.5.1.3.Conducts aperture current.
3.5.1.4.Rinse instrument components between analysis.
3.5.1.5. Carries and Iocuses the sample stream in the Ilow cell to direct the blood cells through
the aperture.
3.5.2.CBC L!ti Rea)ent 7LYSE III Di'' l!ti rea)ent:
3.5.2.1.Rapidly lyses erythrocytes (RBCs, Ireeing hemoglobin (Hgb) and Size oI cellular to a
level that does not interIere with leukocyte (WBC) count.
3.5.2.2.Causes a substantial conversion oI the Hgb to as stable cyanide-containing pigment, the

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 2 oI 12
absorbance oI which is directly proportional to the Hgb concentration over the clinical
range.
3.5.3. Satter Pa9 Rea)ent &!&tem ontain& Er!t-ro!te& II :PA; LYSE< and Sta#ili4e :PA;
LYSE<8 Di'' l!ti rea)ent:
3.5.3.1.Dilutes the blood samples.
3.5.3.2.Rapidly lyses erythrocytes (RBCs).
3.5.3.3.Reduces cellular debris to an insigniIicant level.
3.6.STABLYSE :PA; PRESER%E<8 DI++ PRESER%ATI%E.
3.6.1.Maintaining leukocyte (WBCs) in their near-natural state.
3.6.2. Allows the leukocytes to be diIIerentiated into their subpopulation through the volume,
conductivity and light scatter measurements.
3.7.Reti Pa9 Rea)ent &!&tem ontain& Rea)ent A and Rea)ent B.
3.7.1.REAGENT A, RETIC STAIN
3.7.1.1.Reticulocyte staining solution is a specially Iormulated New Methylene
3.7.1.2.Blue (NMB) dye that stains the reticulum.
3.7.2. REAGENT B, RETIC CLEARING SOLUTION
3.7.1.3. Reticulocyte clearing solution is clearing reagent that removes hemoglobin Irom the
erythrocytes (RBCs) without removing the precipitated dye-RNA complex, keeping
the cell and its membranes intact.
3.8. Clearing Agent
3.8.1. COULTER CLENZ cleaning agent cleans and rinses the internal surIace oI the instrument
components. Daily use prevents protein buildup and eliminates the need Ior routine aperture
bleaching.
3.9. 6UALITY CONTROL:
3.9.1.The quality control program on the Gen S system 2 consists oI running assayed reIerence
material at two shiIts to veriIy the perIormance oI the analyzer. The QC material is divided into
three levels.
3.9.1.1. 5C Cell Control-this is an in-vitro diagnostic reagent consists oI treated, stabilized
human erythrocytes in an isotonic, bacterio static medium. C Cell Control also

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 3 oI 12
contains platelet-sized component, and Iixed erythrocytes o stimulate leukocytes.
S-ed"le:
3.9.1.2. The controls are run two times daily:
Time No. of Levels
Morning (06:00-08:00Hrs) All three levels
vening (16:30-18:00 Hrs) ! levels
Per'ormane C-arateri&ti&:
======================================================================
Parameter Linearit! Ran)e Limit& :Mean di''erene or >
Di''erene8 ?-i-e(er i& )reater
======================================================================
WBC x 103 Cells /uL 0.00 to 100.0 /-0.2 or 3.0
100.1 to 140.0 12.

RBC x 106 Cells /uL 0.00 to 8.00 /-0.05 or 2.0

Hgb g/dl 0.0 to 25.0 /-0.2 or 30

Plt x 10x3 Cells/ul 0 to 1500 /- or 7.0

3.10. ROUTINE SPECIMEN ANALYSIS:
3.10.1. CLOSED %IAL :AUTOMATIC ASPIRATION MODE<:
3.10.1.1. Check reagent levels to ensure adequate levels are present.
3.10.1.2. Check status od Q,C program and iI required, run the controls beIore running
patient samples.
3.10.1.3. II analyzer is in compressor oII mode press CLEAR ALERT.
3.10.1.4. Ensure the instrument is set up Ior the appropriate test.

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 4 oI 12
3.10.!. CHAN.IN. TEST MODE:
3.10.2.1. Go to the analyzer screen.
3.10.2.1.1. Press TEST MODE Irom the MAIN MENU screen oI the SYSTEM
RUN screen.
3.10.2.1.2. Press the key next to the mode oI operation you want:
3.10.2.1.2.1. CBC/DIFF/RETIC Run sample Ior CBC, DiII and Retic
3.10.2.1.2.2. CBC/DIFF Run sample Ior CBC and DiII
3.10.2.1.2.3. CBC ONLY Run sample Ior CBC only
3.10.2.1.2.4. CBC RETIC Run sample Ior CBC and Retic
3.10.2.1.2.5. RETIC ONLY Run sample Retic only
3.10.3. T-e CURRENT MODE di&pla!& t-e te&t mode !o" &elet.
3.10.3.1. Press SYSTEM RUN.
3.10.3.2. The SYSTEM RUN screens display the current test mode.
3.10.3.2.1. Example: MODE : C/D
3.10.3.3. Ensure your specimen have been colleted, stored and mixed properly.
3.10.3.4. Select AUTO ANALYSIS Irom the PROCESSING CONTROL FIELD.
3.10.3.5. Enter the DEMOGRAPHIC oI the patient Irom the SAMPLE ADD ICON Irom the
Workstation.
3.10.3.6. Load the cassettes.
3.10.3.7. Place the cassettes Iirmly and securely into the loading bay on the right side oI the
Diluter the instrument automatically begins cycling the cassettes.
3.10.3.8. AIter the instrument cycles the samples, review the results on the Workstation. Or
VeriIy the result Irom the printout according to the established guidelines.
3.10.". OPEN %IAL ASPIRATION MODE :MANUAL ASPIRATION MODE<
3.10.4.1. Repeat steps 1-7 as Ior CLOSED %IAL ASPIRATION MODE.
3.10.4.2. Enter the sample ID at the Diluter keypad press ID.
3.10.4.3. Press ENTER.
3.10.4.4. Remove the stopper Irom the specimen tube.
3.10.4.5. Immerse the aspirator tip in the tube. The instrument automatically aspirates the

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 5 oI 12
Sample
3.10.4.6. When you hear a beep, remove the tube Irom the aspirator tip. The probe cleaner
retracts the aspirator and automatically cleans it.
3.10.4.7. AIter the instrument cycles the samples, review the sample result on the
workstation or veriIy the result Irom the printout according to the established
guidelines.
3.10.4.7.1. The controls should be run anytime the results or the instrument
perIormance is in @"e&tion.
3.10.4.7.2. Control must be run anytime a new lot number or reagents /diluents are
put to use. And aIter acceptable background check is perIormed.
3.10.4.7.3. Pus Cell Control Normal, Abnormal, Abnormal Low and High- to be
used as reIerence control Ior Ac.t diII. ReIer to assay sheet with each
lot number Ior expected ranges and instructions Ior use. These controls
are to be run daily. They are to be include in the daily start up
procedure. All parameters are to Iiled in the appropriate Ac.t diII
analyzer Iile. This is then printed out at the expiry date oI each 4C Cell
Control lot number the new lot number values are entered.
3.10.4.7.4. Coulter Latron Control and Coulter Latron Primer to be used in
monitoring theVolume, conductivity and light scatter parameters on
Coulter Gen S system 2 only. ReIer to assay sheet Ior expected ranges
and instruction Ior use. All printed out in the appropriate Iile.
3.10.4.7.5. A daily inter instrument comparison is perIormed in the Automated
Hematology between the Coulter analyzers. A manual diIIerential is
perIormed on the same specimen to ensure that the manual and
automated diIIerential results are also in agreement.
3.10.4.7.6. Miscellaneous Controls: Ior procedures not included above, reIer to
procedure manual Ior control requirements.

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 6 oI 12
3.10.#. Open Mode 6C Proed"re
3.10.5.1. Ensure QC specimen is adequately mixed.
3.10.5.2. From the Run Screen, press SPECIMEN TYPE
3.10.5.3. Move the cursor to the desired QC Iile using the arrow key and press 6C
SPECIMEN.
3.10.5.4. II necessary, press CHAN.E SAMPLER to select the open mode.
3.10.5.5. Open the tube and immerse the open sample aspiration in the well-mixed sample.
3.10.5.6. Press the touch plate located behind the probe to start the cycle. Remove the tube
Irom under the aspiration probe when the beep sounds.
3.10.5.7. VeriIy that the results are acceptable.
:Note: out oI control results are displayed in colour).
3.10.5.8. II the results are unacceptable, repeat the run. II the results are still unacceptable,
obtain a new bottle oI the control, be sure that it is warmed and mixed properly and
again repeat the run. II the results are still unacceptable, run the other levels oI
control material. II the results on all levels are unacceptable, troubleshoot
accordingly.
3.10.5.9. Document all unacceptable results on the appropriate log sheet and inIorm the
Senior Technologist and/or Supervisor.
3.10.5.10. When the control results are acceptable, patient samples may be analyzed.
3.11. SAMPLE ANALYSIS PROCEDURE :
3.11.1. +or reprod"i#ilit! &t"die&8 en&"re t-e patient 'or t-e &ample:
3.11.1.1. Is receiving no medication
3.11.1.2. Has normal hematologic parameters, with a WBC count oI 10,000 + 1,000
3.11.1.3. Has normal erythrocyte, leukocyte and platelets morphology and, iI you want to
check the DiII parameters, with DiII values:
3.11.1.3.1. Neutrophils 40 72
3.11.1.3.2. Lymphocytes 17 45
3.11.1.3.3. Monocytes 04 12

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 7 oI 12
3.11.1.3.4. Eosinophils 00 10
3.11.1.3.5. Basophils 00 01
3.11.!. PROCEDURE:
3.11.2.1. Ensure you have enough normal whole blood Irom a single donor Ior 11 cycles.
3.11.2.2. Ensure pneumatics are on
3.11.2.3. Ensure the blood detector is enabled.
3.11.2.4. Ensure the number oI aspiration per tube is set to 1.
3.11.2.5. Ensure the mode oI operation is set to CBC. You can perIorm carryover checks
using other operating modes; however the procedure will take longer and use
excess reagents.
3.11.2.6. Select REPRODUCIBILITY as the processing control on the command center.
3.11.2.7. Select the Reproducibility window to clear out the calues that appear on the results
table.
3.11.2.8. Allocate approximately 1 ml oI the well mixed normal whole blood sample into
one tube.
3.11.2.9. Cycle one sample oI normal whole blood in Automatic aspiration mode.
3.11.2.10. Set the number oI aspiration per tube to 5.
3.11.2.11. Separate the well mixed normal whole blood sample into two tubes.
3.11.2.12. Place the two tubes into consecutive position in a cassette and place the cassette in
the loading bay. The system automatically begins processing the cassette. It
pierces, aspirates, and analyzes the samples.
3.11.2.13. Review the reproducibility results.
3.11.2.14. VeriIy that the CV (CoeIIicient oI Variation) does not exceed the established limits.
II the instruments results exceed these limits, an instruments problem may exist.

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 8 oI 12
REPRODUCIBILITY LIMITS FOR CBC
PARAMETER % CV
WBC
RBC
Hgb
MCV
Plt
MPV
_ 2.5
_ 0.8
_ 0.8
_ 0.8
_ 3.2
_ 5.0
A. POLICY STATEMENT:
".8. All hematology technical staII members should know how to use Cell Gen`s 750 Analyzer and
Coulter Gen`s system in perIorming CBC testing in the context oI adequate quality control measures.
They must be trained on using the operator manual in various instances, e.g. perIorming all allowable
types oI maintenance, entering the quality control material, using diIIerent modes oI sampling, doing
minor troubleshooting. They will be instructed about the department-speciIic system regarding the use
oI analyzer, e.g. how to use the solutions, types oI solutions, regular maintenance schedule, how to
contact the manuIacturing company.
2. PROCEDURE:
#.8. The COULTER GEN. S System is a quantitative, automated hematology analyzer Ior In Vitro
Diagnostic use in clinical laboratories. The GEN S. System provides automated complete blood count,
leukocyte diIIerential and reticulocyte analysis.
#.9. The COULTER VCS (Volume, Conductivity and Scatter) TECHNOLOGY WBC diIIerential and
Reticulocyte using three measurements individual cell volume, high Irequency conductivity and
laser-light scatter.
#.10. The combination oI low-Irequency current and light-scattering technology provides abundant cell-
by-cell inIormation that is translated by the instrument into conventional stained-Iilm leukocyte
categories.

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 9 oI 12
#.11. COULTER IntelliKinetics Application is a management tool Ior the key step oI system
optimization when Iluctuation in external variable in the laboratory, such as temperature.
#.1!. PARAMETERS:
#.1!.1. WBC white blood cell count, NE neutrophil percentage , LY lymphocyte percentage,
MO monocyte percentage, EO eosinophil percentage BA basophile percentage NE#
neutrophil count , BA# basophile count, RBC Red cell count, HGB hemoglobin ,HCT
hematocrit, MCV mean corpuscular volume , MCH mean corpuscular hemoglobin, MCHC
mean corpuscular hemoglobin concentration, RDW red cell distribution curve, PLT platelet
count, MPV mean platelet volume.
#.13. MAINTENANCE PROCEDURES
#.13.1. Maintenance procedures (Ior detailed maintenance procedure reIer to the operator
manual):
5.13.1.1.Dail!
5.13.1.1.1. Start Up
5.13.1.1.2. Run the Auto Clean Cycle
5.13.1.1.3. Clean the Aspiration Needle
5.13.1.1.4. Daily Shutdown
5.13.1.1.5. Keep the external surIace oI the analyzer clean, wipe benches
periodically.
5.13.1.2.Bee9l!
5.13.1.2.1. Replace the Sample Aspiration Peristaltic Pump tubing.
5.13.1.2.2. Clean the air Iilters
5.13.1.2.3. Clean the sample loader and trays
5.13.1.2.4. Run the Auto Clean extended cycle
5.13.1.3.Mont-l!
5.13.1.3.1. Clean the reagent syringes
5.13.1.3.2. Run the extended AUTO-CLEAN Cycle.
5.13.1.3.3. Clean the printer using Iorced air.
#.1". Cali#ration o' t-e .en/& &!&tem 0 LH 123 Anal!4er&.

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 10 oI 12
#.1".1. Gen S System 2 Analyzer is under Iull 24 hrs. service contract. This service contract
provides Ior periodic maintenance to be done quarterly by the
Coulter Engineer, including veriIication oI the calibration Iactors to determine iI a calibration is
warranted.
5.14.1.1.The criteria used to determine the need Ior calibration are:
5.14.1.2.AIter the replacement oI any component the involves dilution characteristics ( Such as
the BSV ) or the primary measurements (Such as the apertures)
5.14.1.3.When controls begin to show evidence oI unusual trends
5.14.1.4.When controls exceed the manuIacturer`s deIined acceptable limits
5.14.1.5.II the average ambient room temperature changes more that 10oF Irom the calibrating
temperature.
5.14.1.6.Once it has been decided that calibration is required, then the Iollowing procedure
should be Iollow.
5.14.1.6.1. PerIorm Reproducibility Check
5.14.1.6.2. PerIorm Carryover Check
5.14.1.6.3. Ensure the instruments is Iunctioning properly
#.1#. CHEC; THE REA.ENT CONTAINER +OR
#.1#.1. SuIIicient quantity
#.1#.!. Not Beyond expiration date
#.1#.3. No precipitates, turbidity, particulate matter, or unusual color.
#.1#.". Proper connection between the Diluter and the Reagent containers.
#.16. CHEC; THE BASTE CONTAINER +OR
#.16.1. SuIIicient capacity
#.16.!. PERFORM DAILY STARTUP

C. RESPONSIBILITY:
6.8.All Lab. Technician and Medical StaII.
1. RE+ERENCES:NIL

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 11 oI 12
D.APPRO%ALS:
Re&pon&i#le Per&on7 Proe&& O?ner:

Specialist, Hematology Section Date
Re(ie?ed #!:

Director, Pathology & Laboratory Medicine Department Date

Director, Medical Department Date
Noted #!:

Director, QI/RM Department Date

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 12 oI 12
Appro(ed #!:

ARMH Deputy President Date

ARMH Vice President Date

ARMH President Date
9. POLICY AND PROCEDURE HISTORY:
Initial PP: IPP-LB-HEM-01-01 Version: 01 Dated:
Replaced By: Version: Dated:
Replaced By: Version: Dated:
Deleted Date: Version: Dated:

Dr. ABDULRAHMAN AL MISHARI HOSPITAL
POLICY and PROCEDURE
INTERNAL POLICY AND PROCEDURE
Department: Pathology and Laboratory Medicine
Poli! Title : LH 750 Analyzer and Coulter Gen`s System Poli! N"m#er $ %er&ion:
IPP-LB-HEM-001-01
E''eti(e Date Re(i&ed Date: Re(i&ion D"e: Replae& N"m#er $
%er&ion:
Applie& To:
Applies to the technical staII oI Hematology Section oI ARMH.
Pa)e N"m#er:
Page 13 oI 12