Hematology

Hematology
• WHAT IS HEMATOLOGY
• WHAT IS ANTICOAGULANT
• DETAILS OF BLOOD CELLS
• PRINCIPLE OF CELL COUNTER
• TYPES OF HEMATOLAGY ANALYZER
• PARAMETERS OF HEMATOLOGY ANALYZER AND ITS SIGNIFICANT
• WHAT IS HISTOGRAME Digestive System
• RBC INDICES EXPLAINANTION
Composition of Blood

Digestive System
 Production of Formed Elements
• Hematopoiesis or hemopoiesis: Process of blood cell production
• Stem cells: All formed elements derived from single population
– Proerythroblasts: Develop into red blood cells
– Myeloblasts: Develop into basophils, neutrophils, eosinophils
– Lymphoblasts: Develop into lymphocytes
– Monoblasts: Develop into monocytes
– Megakaryoblasts: Develop into platelets
Digestive System
Hematopoiesis

Digestive System
 Types of Blood Cells
• There are three kinds of blood cells.

Digestive System

Leukocyte Erythrocyte Platelet

Size: 30-400fl 30-250fl 2-25fl
6 9 6
Amount: 4-10X10 /ml 3.5-5.5X10 /ml 100-300X10 /ml
Luckocytes (WBC)

Granulocyt
WB
e Digestive System
DefendC
the body
against infectious
disease and
foreign materials
Blood cells

Lymph Baso Mono Eosino

Neutro

Digestive System

Lymphocyte Mid-sized Cell Neutrophilic Granulocyte

 Erothrocytes (RBC)
• The function of RBC is almost completed by hemoglobin (HGB).

Transport oxygen and carbon dioxide

Digestive System
Plateletes

Coagulation or clotting

Digestive System
 What is Anticoagulant

Anticoagulant is the chemical which has property to stop the clotting of blood when it mixed in
optimum quantuty.

there are many types of anticoagulants i.e.: happrin / EDTA / K2EDTA / K3 EDTA

Both K2EDTA and K3 EDTAcan be used as anticoagulant for whole blood. But there is
Digestive
some difference. The blood can be stored for 8System
hours With K2EDTA , and only 4hours with
K3 EDTA. So usually we suggest K2EDTA .
 Principle of Hematology Analyzer

Electrical impedance methode

Volumetric Metering

Digestive
Colorimetric System
Method

Reagents
 Electrical Impedance Method
• WBC, RBC and PLT are counted and sorted by the Electrical Impedance Method, just
Coulter Principle. This method is based on the measurement of changes in electrical
impedance produced by a particle passing through an aperture.

Digestive System
Digestive System
 Digestive System

The amplitude of each pulse is proportional to the size of each blood cell.
Pulse Graph
1 、 blood cell of Same size, go through the aperture
by constant speed.

2 、 blood cell of Same size, go through the aperture

by variable speed.
Digestive System

3 、 blood cell of different size, go through the

aperture by variable speed.
Digestive System
• From Pulse Graph to Histogram

Digestive System
From Pulse Graph to Histogram

Digestive System
Digestive System
Digestive System
Digestive System
Volumetric Measurement
This analyzer uses a volumetric metering unit to control the count cycle
and to ensure that a precise volume of sample is analyzed.

Digestive System
Colorimetric Method

• HGB is determined by the colorimetric method.

• Lyse will release HGB when RBC was broken down and react with HGB to
generate a mixture for HGB measurement.
• The mixture has the biggest absorbance at 540nm.

I0 It

Digestive System

L
Colorimetric Method

Light Absorbing Detector Signal processing

source container Display

Digestive System
LED light Bath Photosensor
Reagents

Reagents are one part of the system.

• Lyse
Break down RBC; Release HGB and react with HGB to generate a mixture for HGB
measurement
Help the differential of WBC.
Digestive System
• Diluent
Dilute the blood sample;
Provide a conductive environment for counting and environment which is similar to the
plasma to maintain the size of blood cell Clean the tube and bath.
 Reagents

• Rinse / Detergent
Clean the bath and metering tube.

• Cleanser
An enzyme-based cleaning solution used toSystem
Digestive clean the tube and bath.

• Probe cleanser
An alkaline cleaning solution used to clean the tube and bath.
 What is ”cyanide free reagent”?
As you may have known, the recommended method for HGB measurement is
HiCN (hemiglobincyanide) by ICSH (international committee for standardization
in hematology.) With this method, KCN (potassium cyanide) is existed, which is of
serious toxicity.

Thus, usually when we say cyanide free lyse, it means no KCN is existed. For
Meril cyanide free lyse, FeCN2 (nontoxic) is used. That's why our reagents are
safe. On our reagent label, you can find theSystem
Digestive main components.
Hematocrit (HCT)

• The hematocrit (HCT), or packed cell

volume (PCV) represents the proportion
of blood composed of red blood cells,
expressed as % (vol/vol). It is the
quickest and most accurate measure of
the red cell component of blood.
• Traditionally, it is determined by
measuring the height of the red cell
column in a microhematocritDigestive
tube System
following centrifugation (see fig.
Automated analyzers calculate the HCT
by multiplying the red cell count and the
mean red cell volume, both of which are
measured directly by the machine.
Mean Cell Volume (MCV)

• The mean cell volume indicates the volume of the "average" red cell in a sample.
• It is expressed in femtoliters (fl; 10-15 liters).
• The normal range is 80 - 94 fL.
• Normocytic refers to blood with a normal MCV
• Red cell populations with the MCV above the reference interval are
termed macrocytic
Digestive System
• Red cell populations with the MCV below the reference interval are termed
microcytic.
Mean corpuscular hemoglobin (MCH)

• The MCH represents the mean mass of hemoglobin in the RBC.

• It is expressed in the mass unit, picograms (pg, 10 -12 gram).
• The value is determined by the formula,
(Hemoglobin [in g/dL] x 10 ÷ (RBC count [in millions/µL])
• The normal range (27 - 31 pg).
• Since small cells have less hemoglobin than large cells, variation in the MCH tends
Digestive System
to track along with that of the MCV.
Mean corpuscular hemoglobin concentration (MCHC)

• This is the mean concentration of hemoglobin in the red cell.

• Since whole blood is about one-half cells by volume, and all of the hemoglobin is
confined to the cells, you would correctly expect the MCHC to be roughly twice
the value for hemoglobin in whole blood
• It is expressed in g/dl.
• The normal range is 32 - 36 g/dL.
• Digestive
The value is calculated using the System
formula,
Hemoglobin [in g/dL] ÷ hematocrit [in L/L]
• Cells with normal, high, and low MCHC are referred to as normochromic,
hyperchromic, and hypochromic, respectively.
• These terms will have importance in anemia classification.
The Red Blood Cell Distribution Width (RDW)

• It is a measure of deviation of the volume of RBCs

• However, "width" refers to the width of the volume curve (distribution width), not
the width of the cells
• Usually red blood cells are a standard size of about 6–8 μm.
• Certain disorders, however, cause a significant variation in cell size.
• Higher RDW values indicate greater variation in size.
• Digestive
Normal reference range in human System
red blood cells is 11–15%.
• Mathematically the RDW is calculated with the following formula:

RDW = (Standard deviation of MCV ÷ mean MCV) × 100.

• The determination of the RDW-SD on instruments is an actual measurement of the

width of the erythrocyte distribution curve. This measurement is performed at a
relative height of 20 % above the baseline.
Digestive System
• The wider the curve is spread by erythrocytes of different sizes, the higher the
RDW-SD value will be.
• Reference values:
female: 36.4 – 46.3 fL
male : 35.1 – 43.9 fL
RDW-CV

• The RDW-CV is calculated from the formula:

• RDW-CV = 1SD x 100
MCV
• 1SD reflects the size variation of the erythrocytes round the mean. As the 1SD is
divided by the MCV, the RDW-CV also depends on the mean size (MCV) of the
erythrocytes. Digestive System
• Reference values:
Female: 11.7 – 14.4%
Male: 11.6 – 14.4 %
The RULE of Three

HGB = RBC X 3
HCT = HGB X 3

RBC = 3.0 x 1012 3x3=9

Hb = 9.2 g/dL Digestive System

3 x 9.2 = 27.6

Hct = 28%
White Blood Cells

5 different types of WBCs

Normal Values % Absolute

Neutrophils 40 – 70 1800 – 7500

Eosinophils 0 –System
Digestive 6 0 – 600

Basophils 0–1 0 – 100

Lymphocytes 20 – 45 900 – 4500

Monocytes 2–8 90 - 1000

Types of Hematology Analyzer

3 Part or 3 diff Analyzer

- Single Chamber
- Dual Chamber

5 Part or 5 diff Analyzer

- Basic Model
- With Retics and nRBC count
Digestive System
How the all 23 perameter Reported

The Parameter derived by 3 methods

- By Actual measurement
- By Histogram
- By Claculation

Digestive System
Results come from Measuring
White Blood Cell WBC
Lymphocyte Lymph# Red Blood Cell RBC
Mid-sized cell Mid# Hemoglobin HGB
Granulocyte Gran# Hematocrit HCT
Lymphocyte % Lymph% Mean Cell (erythrocyte) Volume
Mid-sized cell% Mid% MCV
Granulocyte% Gran% Mean Cell (erythrocyte) Hemoglobin
MCH
Platelet PLT Mean Cell (erythrocyte) Hemoglobin
Mean Platelet Volume MPV Digestive System
Concentration MCHC
Red Blood Cell (erythrocyte)
Platelet Distribution Distribution Width
Width PDW Coefficient of variation RDW-CV
Plateletcrit PCT Red Blood Cell (erythrocyte)
Platelate Large cell Distribution Width Standard
Ratio P-LCR deviation RDW-SD
Results come from Histograme
White Blood Cell WBC
Lymphocyte Lymph# Red Blood Cell RBC
Mid-sized cell Mid# Hemoglobin HGB
Granulocyte Gran# Hematocrit HCT
Lymphocyte % Lymph% Mean Cell (erythrocyte) Volume
Mid-sized cell% Mid% MCV
Granulocyte% Gran% Mean Cell (erythrocyte) Hemoglobin
MCH
Mean Cell (erythrocyte) Hemoglobin
Platelet PLT Concentration MCHC
Mean Platelet Volume MPV DigestiveRed
System
Blood Cell (erythrocyte)
Distribution Width
Platelet Distribution Coefficient of variation RDW-CV
Width PDW Red Blood Cell (erythrocyte) Distribution
Plateletcrit PCT Width Standard
Platelate Large cell deviation RDW-SD
Ratio P-LCR
Results come from Calculation
White Blood Cell WBC
Lymphocyte Lymph# Red Blood Cell RBC
Mid-sized cell Mid# Hemoglobin HGB
Granulocyte Gran# Hematocrit HCT
Lymphocyte Mean Cell (erythrocyte) Volume
%Lymph% MCV
Mid-sized cell% Mid% Mean Cell (erythrocyte) Hemoglobin
Granulocyte%
Platelet Gran%
PLT MCH
Mean Platelet Volume MPV Mean Cell (erythrocyte) Hemoglobin
Platelet Distribution DigestiveConcentration
System MCHC
Width Red Blood Cell (erythrocyte) Distribution
PDW Width
Plateletcrit Coefficient of variation RDW-CV
PCT Red Blood Cell (erythrocyte) Distribution
Platelate Large cell Width Standard
Ratio P-LCR deviation RDW-SD
 Installation Requirements

Environment

Operating temperature: 15 ℃ to 30 ℃.
Relative humidity: 30% to 85%.
Atmospheric pressure: 70 kPa to 106 kPa.
free as possible from dust, mechanical vibrations, loud noises, and electrical i
nterference.
Digestive System
Do not place near brush-type motors, flickering fluorescent lights, and electric
al contacts that regularly open and close.
Do not place in direct sunlight or in front of a source of heat or drafts.
Installation Requirements

Space

At least 40 cm on each side, which is the preferred access to perform service

procedures

At least 10 cm behind for cabling and ventilation

Digestive System
There is enough room on the countertop to ccommodate waste bottle
Installation Requirements

Power

Check the availability of a power outlet that is

 a female receptacle;
 single phase with ground (confirmed third-wire earth ground);
 voltage of zero curve to ground should be ≤5V
Voltage: 100 VAC - 240 VAC;
Frequency: 50/60 ±1 Hz Digestive System
Fuse: 250V T4A
Better to have an UPS: 500W
Properly grounded socket or connect the grounding pole at the back of the analyzer to
the ground.
 Installation Procedures

BOX for CelQuant 3i Open the box

Digestive System
Remove the Remove foam covers
instrument from box Place it on flat
very carefully surface

Digestive System
Remove shrink Connect all Remove plastic
wrap tubing's and wrap from top of
wires chamber cover and
cut cable tie

Digestive System
 Install Printer Paper

1.Open the door of the Thermal Printer.

2.Uplift the bar of the Thermal Printer.
3.Insert the paper into the paper entry.
4.Push the bar of the Thermal Printer.
Digestive System
5.Close the door.

Note: The thermo-side should faces up.

 Press Service Option Press PRIME LISE once
Switch ON the
priming is done again
instrument
press Prime Lyse. Repeat
same procedure with
Diluent and Rinse

Digestive System
After Priming is done give 3-4 Homescreen
Blank

Digestive System
Now Run Sample 5-6 times Check Result and it’s
Repeatability

Digestive System
 Repeatability Test
• Good repetition, poor accuracy Recalibrate

• 8.9, 9.0, 8.9, 8.9, 8.8 (reference value: 9.6+0.5)

Troubleshooting

• Poor repetition, poor accuracy

• Digestive
8.5, 9.4, 10.1, 8.0, 10.5 System

Perfect

• Good repetition, good accuracy

• 9.6, 9.7, 9.7, 9.6, 9.5

Comparison of instruments
COMPANY NAME MERIL SYSMEX NIHON KOHDEN

IMAGE

Digestive System
TECHNICAL SPECIFICATION
THROUGHPUT 35 SAMPLES/HOUR 60 SAMPLES/HOUR 60 SAMPLES/HOUR
WORKING PRINCIPLE electrical resistant electrical resistant electrical resistant
SAMPLE venous,capillary,pre diluated venous,capillary,pre diluated venous,capillary,pre diluated
MEASUREMENT PRINCIPLE electrical empedance electrical empedance electrical empedance
PARAMETERS 20 parameters with three histogram 20 parameters with three histogram 19 parameters with three histogram
SAMPLE VOLUME 9.8ul&20ul in pre-diluated mode 10ul&20ul in pre-diluated mode 30ul/10ulor20ul in pre-diluated mode
APERTRE SIZE wbc-100ul/rbc&plt-70ul

REAGENT REQUIREMENT(PER SAMPLE) diluent-20ml/rinse-5ml/lyse-0.3ul

QUALITY CONTROL 4QC/day&3 month QC data stored 4QC/day&3 month QC data stored
SCREEN lcd colour touch screen lcd colour touch screen lcd colour touch screen
PRINTER thermal printer thermal printer thermal printer
STORAGE 10,000 TEST with histogram 35,000 TEST with histogram 10,000 TEST with histogram
 HUMAN CYPRESS ABBOTT
COMPANY NAME

IMAGE

TECHNICAL SPECIFICATION
30/60 SAMPLES/HOUR 30 SAMPLES/HOUR 57 SAMPLES/HOUR
THROUGHPUT
electrical resistant electrical resistant electrical resistant
WORKING PRINCIPLE Digestive System
venous,capillary,pre diluated venous,capillary,pre diluated venous,capillary,pre diluated
SAMPLE
electrical empedance electrical empedance electrical empedance
MEASUREMENT PRINCIPLE
PARAMETERS
22 parameters with three histogram 20 parameters with three histogram 16 parameters with three histogram
SAMPLE VOLUME
25ul/25ul in pre-diluated mode 25ul/50ul in pre-diluated mode 9.8UL/ in pre-diluated mode
APERTRE SIZE
REAGENT REQUIREMENT(PER
70ul-RBC,PLT,WBC ?????
SAMPLE) ?????
QUALITY CONTROL 6 level controls 3 level controls 3 level controls
SCREEN lcd colour touch screen lcd colour touch screen lcd colour touch screen
PRINTER thermal printer thermal printer ?????
STORAGE 10000 TEST with histogram 10000 TEST with histogram 1500 TEST with histogram
USP

* Less reagent consumption

* Dual Chambers

* Larger Display

* Embedded External Printers

Digestive System

* Pre diluted mode

Thank you
Digestive System