YUMIZEN H550

DIAGOMET
AUTOMATED BLOOD HAEMATOLOGY
LABORATORIES, DELHI
COUNT ANALYSER

Issue No: 01 Date: Page 1 of 6

1. Purpose: Quantitative enumeration of different formed elements (cells) of blood and

Hemoglobin in whole blood by HORIBA YUMIZEN H550 an automated Hematology
cell counter by the principle of Flowcytometry & Impedance technology.
HORIBA YUMIZEN H55O automated Hematology System enumerates the following
parameters:
Total RBC count
Total WBC count
Differential WBC Count – Both absolute and Percentage
Hemoglobin
Hematocrit (HCT)
Mean Corpuscular Volume (MCV)
Mean Corpuscular Hemoglobin (MCH)
Mean Corpuscular Hemoglobin Concentration (MCHC) &
Read Cell Distribution Width (RDW CV & SD)
Platelet count (PLT)
Platelet Cell Distribution Width (PDW)
Plateletcrit (PCT)
Platelet Large Cell Ratio (PLCR)
Platelet Large Cell Count (PLCC)
Mean Platelet Volume (MPV)

It is the enumeration of cellular elements of blood, evaluation of red cells indices and
determination of cell morphology by means of stained smears. The various parameters generated
from the automated hematology analyzer are of clinical importance to differentiate between
diseases of hematological abnormalities. Reference values vary across the life cycle and between
sexes and differ in different age groups and in various disease conditions.

2. Principle:
WBC
The WBC detection principal is based on the Double Hydrodynamic Sequential System
“DHSS” which allows a linear flow of the cell through the light path (Horiba Medical Patent
technology)

Designation Signature
Prepared by: Senior Lab Technician
Reviewed & approved by: Quality Manager
Issued by: Technical Manager
 YUMIZEN H550
DIAGOMET
AUTOMATED BLOOD HAEMATOLOGY
LABORATORIES, DELHI
COUNT ANALYSER

Issue No: 01 Date: Page 2 of 6

Measurement of impedance variation generated by the passage of WBC cells through a

calibrated micro aperture and Double Hydrodynamic Sequential system. The 20µl of specimen is
diluted in 1.4 ml of Whitediff reagent and keep for 22 seconds incubation for 37ºC. In this
reaction destroy RBC membrane and stabilize WBC to prepare the cell identification in the
cytometer.

 In the optical chamber during the acquisition of the LMNE matrix. (DHSS)

RBC/PLT
Measurement of impedance variation generated by the passage of RBC cells through a
calibrated micro aperture. The specimen is diluted in ABX Diluent (current conductor) and pulled
through the calibrated micro-aperture. Two electrodes are placed on either side of the aperture.
Electric current passes through the electrodes continuously. When the cell passes through the
aperture, electric resistance between the two electrodes increases proportionately to the cell
volume.

HB

Whitediff is a reagent used for erythrocyte lysis and cyanide-free determination of

hemoglobin. By addition of agent of lysis, hemoglobin is released. All the heme iron is oxidized
and stabilized. Oxidation resulting complexes are quantified by spectrophotometry at a
wavelength of 555 nm.

HCT

All the RBC pulses are grouped into various sizes. Each group pulse height is then
averaged. All the pulse height averages are then averaged one final time for a mean average of
all the RBC pulse heights. This function is a numeric integration of the MCV, The HCT results
are given as a percentage of this integration
.
MCV: calculated from RBC and HCT
MCH: Calculated from Hb and RBC
MCHC: calculated from Hb and Hct
RDW: Calculated from deviation of RBC Distribution Width

Designation Signature
Prepared by: Senior Lab Technician
Reviewed & approved by: Quality Manager
Issued by: Technical Manager
 YUMIZEN H550
DIAGOMET
AUTOMATED BLOOD HAEMATOLOGY
LABORATORIES, DELHI
COUNT ANALYSER

Issue No: 01 Date: Page 3 of 6

3. Performance specification:
Measurement Minimum detection
Linearity
Range range
WBC 0.0 – 300 x 103 0.5 – 15.0 x 103
0.3 (300 Cells / cu mm)
( X 103 Cells / μL) cells/μL cells/Μl
RBC 0.0 – 8.00 x 106 0.2 – 6.00 x 106 0.2 ( 0.2 million cells /
( X 106 Cells / μL) cells/μL cells/μL cumm)
Platelets 0.0 – 2500 x 103 10 – 500 x 103 10 (10,000 cells / cu
( X 103 Cells / μL) cells/μL cells/μL mm)
Hb (g/dL) 0.0 – 24.0 g/dL 2.5 -18.0 g/dL 2.5 g /dL

Hct (%) 67 11.6 – 35.0 11.6

4. Primary Sample:
1. Use whole blood as sample collected in K3 EDTA
2. Collect 2 ml blood in lavender top vacutainer tube (Containing 3.6 mg K 2 EDTA) or
in tubes containing 40 μL of 10 % K2 EDTA per 2 ml of blood).
3. Mix the specimen thoroughly but gently in a hemomixer for 3 – 5 minutes
immediately prior to aspiration of the sample by the machine/ by 3- 5 gentle inversion
and rotation between the palms of hands.
4. Process the samples within 4 hours of collection.
5. Do not use samples which are clotted, containing fibrin microclots, decomposed or
contaminated sample.
6. Reject clotted samples and samples without ID.
5. Type of container and additive: Use a lavender color topped vacutainer tube
(containing 3.6 mg K3 EDTA)
6. Consumables/ Reagents:
1. Horiba Series reagent system,
2. ABX diluents 20L
3. Whitediff 1L
Designation Signature
Prepared by: Senior Lab Technician
Reviewed & approved by: Quality Manager
Issued by: Technical Manager
 YUMIZEN H550
DIAGOMET
AUTOMATED BLOOD HAEMATOLOGY
LABORATORIES, DELHI
COUNT ANALYSER

Issue No: 01 Date: Page 4 of 6

4. ABX Cleaner 1L
7. Equipment: HORIBA YUMIZEN H500
8. Step by Step Procedure: HORIBA YUMIZEN – VI PART

Switch on CPU

Switch on the Instrument

“Login”

Startup

Control Run

QC File - Select File

Run Control Level-1

Run Control Level-2

Run Control Level-3

Go For Manual Entry

Sample Analysis

START (Enter)

Run the Sample

Press GP Report for Print

Or Press HC For Host Communication

Go For Shutdown Menu Execute

Switch Off the analyzer

1. Switch on the instrument by pressing the ON/OFF Switch.

Designation Signature
Prepared by: Senior Lab Technician
Reviewed & approved by: Quality Manager
Issued by: Technical Manager
 YUMIZEN H550
DIAGOMET
AUTOMATED BLOOD HAEMATOLOGY
LABORATORIES, DELHI
COUNT ANALYSER

Issue No: 01 Date: Page 5 of 6

2. Wait for Approximately 3 min for the initialisation process to be completed. (Only
Sart-up)
9. Quality Control Procedure:
1. Run all three levels of an assayed QC sample (Horiba Three Level Quality Control
samples - Low, Normal and High) and verify if the values are close to the Mean and
within the reference range mentioned in the QC product insert.
2. If the QC is not within the reference range even for any one level of QC, perform
another cleaning procedure and rerun that level of QC.
10. Reference Range:

Parameter Reference Range

WBC count Adults 4000-11,000 cells/cu.mm

Male : 4.28 to 5.79 x 106cells/cu.mm
RBC count
Female: 3.93 to 5.19 x 10 6 cells/cu.m
Men: 13.4 – 16.7g/100ml
Haemoglobin
Women 11.5-15.1 g/ dl
Platelet count 150,000 –450,000/cu.mm
Male: 39.2 – 48.6 %
Haematocrit
Female 34.4 – 44.6%
MCV 78-97 fl

MCH 27-32 gm/dl

MCHC 32.0-36.0%
11. Potential Sources of variability :
1. The Following factors will affect cell counting in an automated hematology analyzer
2. Visible clots Fibrin micro clots or cryoprecipitate in the specimen
3. Inadequate lysis of RBC
4. Inadequate dilution of specimen and inadequate calibration of the analyzer
5. Non Homogenous suspension of blood cells in the diluting fluid
Designation Signature
Prepared by: Senior Lab Technician
Reviewed & approved by: Quality Manager
Issued by: Technical Manager
 YUMIZEN H550
DIAGOMET
AUTOMATED BLOOD HAEMATOLOGY
LABORATORIES, DELHI
COUNT ANALYSER

Issue No: 01 Date: Page 6 of 6

6. Deterioration of diluting solution.

7. Presence of dust particles in the diluent. Ensure the diluents and buffer solutions
used are free of dust particles especially for platelet counting.
8. Partial or total obstruction of the aperture.
9. Wrong threshold setting of the instrument
10. Carry over from one to next measurement
11. Fluctuation of electric current

12. Reference / Source Document:

1. User Manual of HORIBA YUMIZEN H550

Designation Signature
Prepared by: Senior Lab Technician
Reviewed & approved by: Quality Manager
Issued by: Technical Manager