LAB - PACKED CELL VOLUME TEST (PCV)

Introduction:

Haematocrit is also known as Packed Cell Volume (PCV). The packed cell volume (PCV) is the measure of the ratio of the volume occupied by the red cells to the volume of whole blood in a sample of capillary, venous, or arterial blood. The word hematocrit means “to separate blood”, which underscores the mechanism of the test because the plasmas and blood cells are separated by centrifugation.

Objective:

To measure the ratio of the volume occupied by red blood cells to the volume of whole blood in a sample of venous blood.

Equipments:

1. Microhaematocrit tube
2. Micorhaematocrit centrifuge
3. EDTA venous blood
4. Plastercene
5. Microhaematocrit reader

Procedure:

1. The blood is mixed well.
2. The blood is filled into the haematocrit tube until it is 4/5 of the blood.
3. The other end of the capillary tube is sealed with plastercene.
4. The capillary tube is placed into the microhaematocrit centrifuge.
5. The capillary tube is centrifuged for 5 minutes for 12000 rpm.
6. Read and record the results by using microhaematocrit reader.

Questions:

1. Define haematocrit.
2. What is the other term for haematocrit?
3. Name three layers of centrifuged blood.
4. Why it is important to read the microhaematocrit soon after the centrifuge stops?

LAB - WHITE BLOOD CELL (LEUCOCYTE) COUNTING

Introduction:

A white blood cell count estimates the total number of white cells. It is important in the diagnosis of disease especially when accompanied by a differential white cell count.

Two method to evaluate total number of leucocytes

1. Manual method
2. Automation technique

Reagents:

Dilution solution:

Tuerk dilution solution: 2% glacial acetic acid

®       Glacial acetic acid                  

®       Distilled water

®       Gentian violet

Principle:

Acetic acid will lyse RBC, so WBC only can be count.

Procedure:

1. Using a red blood cell Thoma pipette draw well mixed blood and fill till the 0.5 mark. Clean the tip of the tube.
2. Now draw WBC diluting fluid to the 11 mark which is 1:20 dilution.
3. Mix well for 3 minutes.
4. Discard the first three to four drops of mixture and gently fill in the mixture into the Neubauer chamber.
5. Under 10X magnification, scan to ensure even distribution.
6. Count the red blood cells using 40X objective in the 80 smallest squares as indicated in the diagram of the chamber.

Questions:

1. What are the factors that may occur for the wrong counting of white blood cells?
2. Define leucocytosis.
3. What is the normal white blood cell range for adults and babies?

LAB - PLATELET COUNTING

Equipments:

1. Formal citrate dilution solution
2. Neubauer chamber
3. Red blood cell Thoma pipette
4. Microscope
5. Blood sample

Reagents:

Dilution solution: Formal citrate

1. 1% (l/l) Formalin (40% Formaldehyde) in 3% Trisodium Citrate.
2. 1-2 drops of Brilliant Cresyl blue (1% in isotonic saline) in each litre of formal citrate solution.

Procedure:

1. Using a red blood cell Thoma pipette draw well mixed venous blood or capillary blood and fill till the 0.5 mark. Clean the tip of the tube.
2. Draw diluting fluid to the 101 mark.
3. Shake and mixed well for 5 minutes.
4. Discard the first three to four drops of mixture and gently fill in the mixture into the Neubauer chamber.
5. Under 10X magnification, scan to ensure even distribution.
6. Count the red blood cells using 40X objective in the 80 smallest squares as indicated in the diagram of the chamber.

Normal range:

150-400 x 10⁹ / L

Questions:

1. List TWO conditions for thrombocytopenia
2. List TWO conditions for thrombocytosis
3. State the normal range for platelets.

LAB - RED BLOOD CELL COUNT

Introduction:

Red blood cell counting is an important diagnostic test towards observation of peripheral blood film. Changing in total number of red blood cells is related to physiological or pathological method. Increase in red blood cell is related to polycythaemia cases while decrease of total number of red blood cells is related to anaemia cases.

Equipments:

1. Formal citrate dilution solution
2. Neubauer chamber
3. Red blood cell Thoma pipette
4. Microscope
5. Blood sample

Reagents:

Dilution solution: Formal citrate

1. 1% (l/l) Formalin (40% Formaldehyde) in 3% Trisodium Citrate.
2. 1-2 drops of Brilliant Cresyl blue (1% in isotonic saline) in each litre of formal citrate solution.

Procedure:

1. Using a red blood cell Thoma pipette draw well mixed venous blood or capillary blood and fill till the 0.5 mark. Clean the tip of the tube.
2. Draw diluting fluid to the 101 mark.
3. Shake and mixed well for 5 minutes.
4. Discard the first four drops of mixture and gently fill in the mixture into the Neubauer chamber.
5. Under 10x magnification, scan to ensure even distribution.
6. Count the red blood cells using 40X objective in the 80 smallest squares as indicated in the diagram of the chamber.

Normal range:

Men                 :           4.5-6.5 x 10¹² / L

Women            :           3.8-5.8 x 10¹² / L

Babies             :           4.0-6.0 x 10¹² / L

Questions:

1. State FOUR (4) sources of error and indicate if the cell count would be falsely increased or decrease.
2. What is the purpose to dilute the blood with formal citrate?

LAB - PREPARATION OF CYANMETHEMOGLOBIN STANDARD GRAPH

REAGENTS AND EQUIPMENTS

1.         Drabkin’s solution

2.         Test tubes

3.         Micropipette

4.         Spectrophotometer

Specimens

EDTA / capillary blood

Preparation of cyanmethemoglobin standard graph

Procedure

1.         Prepare and label 5 test tubes.

2.         Dilution factor use is 1:300 if 6ml of Drabkin’s solution are used. The given standard 

            concentration (cuanmethemoglobin) is 57.2 g/ dL.

3.         The contents are mixed thoroughly.

4.         Read the absorbane of each solution at 540 nm.

5.         Record the results and prepare a standard graph.

Table for cyanmethemoglobin standard graph

Number	Drabkin’s solution	Standard Cyanmethemoglobin solution	Standard Cyanmethemoglobin concentration	Absorbance reading
1	6.0	0.0
2	5.0	1.0
3	3.0	3.0
4	1.0	5.0
5	0.0	6.0

Procedure

1.         Transfer 20µl of blood to 6.0 ml Drabkin’s solution

2.         Mix well and leave in a dark place for 10 minutes

3.        Read the absorbance of each solution at 540nm using a spectrophotometer and use Drabkin’s solution as ‘Blank’.

Tests tube	Blood A	Blood B
Drabkin’s solution	6.0 ml	6.0 ml
Blood sample	20µl	20µl
Standard cyanmethemoglobin concentration	X	Y
Absorbance reading	a	b

QUESTIONS

1.                List 3 errors that can interfere with haemoglobin determinations.

2.               State the normal range for haemoglobin for Male and Female.

3.              What is the wavelength used to measure haemoglobin manually on the spectrophotometer?

PERFORMING VENIPUNCTURE ON BABY

•         Recommended location - the heel.

•         Prewarming the infant's heel.

•         Hold the baby's foot firmly to avoid sudden movement.

•         Using a sterile blood lancet, puncture the side of the heel.

•          Do not use excessive pressure because the blood may become diluted with tissue fluid.

•         Fill the capillary tubes as needed.

•         When finished, elevate the heel, place a piece of clean, dry cotton on the puncture site, and hold it in place until the bleeding has stopped.

•         Dispose the lancet in appropriate sharps container.

Hematology

Hematology is a study of the formation and maturation of blood cells. Many hematologic diseases involve failure of maturational mechanisms, abnormal cell differentiation, or failure of regulation of cell proliferation. The pathophysiology of many blood disorders can be explained by study of the proliferative capacity and the degree of differentiation seen when marrow cells are studied microscopically and cultured. The degree of demand, the adequacy of reverses, and the proliferative and differentiation capacity of the bone marrow govern the number of normal and abnormal cells seen in the peripheral blood and bone marrow in certain diseases.

This equips the Medical Laboratory Technologists with theoretical knowledge and technical skill to investigate and diagnose various types of anaemia including iron deficiency anaemia, folae/B12 deficiency anaemia, thalassemia and abnormal haemoglobins. This enables the Medical Laboratory Technologists to understand leukaemia based on classification, aetiology, clinical manifestations and various features.Medical Laboratory Technologists are equipped with the knowledge and technical skill to detect and diagnose this disease through Full Blood Picture, bone marrow study and cytochemistry and also exposes the theory of hemostasis and methods to investigate bleeding and clotting disorders.

Main Reference Textbooks:
Clinical Laboratory Haematology, April 2003, Shirlyn Mckenzie, Prentice Hall


Essential Haematology 5th Edition 2006 A.V.Hoffbrand


Additional Reference Materials:
Practical Haematology, Sir John Dacie & Lewis, 10th Edition, 2004


Anderson’s Electronic Atlas of Hematology 2002 Shauna C Anderson, Lippincott

Determination of Glucose-6-Phosphate Dehydrogenase in Erythrocytes

The hexose monophosphate shunt is essential in erythrocyte metabolism, liver, adipose tissue and kidney metabolism. The regulation of pentose phosphate is a dehydrogenation step of glucose-6-posphate by G6PD. A decrease in G6PD enzyme in erythrocyte will cause a reduction in NADPH production and haemolysis occurs. G6PD deficiency is an inherited x-linked recessive disease. Its determination is very important since the deficiency of it can cause haemolytic anaemia and neonatal jaundice.

G6PDH is an enzyme of the pentose phosphate pathway that catalyzes the reaction below:

Glucose-6-Phosphate + NADP+ <=> 6-Phosphogluconolactone + NADPH

Individuals whose G6PDH is partly inactive are prone to oxidative stress. This arises because NADPH produced by the pentose phosphate pathway is required for reduction of glutathione, which, in turn, helps reduce peroxides. In the absence of sufficient NADPH, glutathione remains oxidized and peroxides cannot be neutralized.

In this of experiment we have carried out determination of glucose-6-phosphate dehydrogenase in erythrocyte which is a significant enzyme in pentose phosphate pathway. When G6PD is present in erythrocytes at normal level, it can act on glucose-6-phosphate and catalyses NADP+ to NADPH which in turn will emit UV light, absence of UV light indicates a decrease in the enzyme level. A decreased level of G6PD enzyme in erythrocytes will cause reduction of NADPH production and consequently hemolysis will occur (hemolytic anemia).

A mutation present in some populations causes a deficiency in glucose-6-phosphate dehydrogenase with consequent impairment in the generation of NADPH. This impairment is manifested as red cell hemolysis when the susceptible individual is subject to oxidants such as the antimalarial primaquine, aspirin, or sulfonamides or when the susceptible individual eats fava beans (Vicia fava).

The Pentose phosphate pathway supplies RBC with NADPH to maintain the reduced state of glutathione. The inability to maintain reduced glutathione in RBC’s leads to increased accumulation of peroxides, predominantly H202 that in turn results in a weakening of the cell wall and concomitant hemolysis.Therefore, any defect in the production of NADHP could, therefore, have profound effects on erythrocyte survival.

Hemolytic anemia is the red blood cell (RBC) plasma membranes rupture prematurely. The released haemoglobin pours into the plasma and may damage the filtering unit (glomeruli) in the kidneys. The condition may result from inherited defects such as abnormal red blood cell enzymes, or from outside agents such as parasites, toxins, or antibodies from incompatible transfused blood.

The symptom of hemolytic anemia is the reduced number of RBC’s or a decreased amount of haemoglobin in the blood. The person feels fatigued and is tolerant of cold, both of which are related to lack of oxygen needed for ATP and heat production. Also, the skin appears pale due to the low content of red-colored haemoglobin circulating in skin blood vessel. That is why G6PD determination or verification is very significant especially for new born babies as a screening test.

Hemoglobin Determination Using Cyanmethemoglobin Method

Principle

In a reagent solution the ferrous ions (Fe2+) of hemoglobin are oxidized to the ferric (Fe3+) state by potassium ferricyanide to form methemoglobin. Methemoglobin subsequently reacts with the cyanide ions provided by potassium cyanide to form cyanmethemoglobin. The amount of cyanmethemoglobin can be measured spectrophotometrically at a wavelength of 540 nm on a spectrophotometer and compared to known hemoglobin standards in order to determine the hemoglobin concentration of the unknown sample. Hemoglobin, the main component of the RBC, transports oxygen to and CO2 from the body’s tissues. Hemoglobin in circulating blood is a mixture of hemoglobin, oxyhemoglobin, carboxyhemoglobin and minor amounts of other forms of this pigment. It is necessary to make a stable derivative involving all forms of hemoglobin in the blood in order to measure this compound accurately. The cyanmethemoglobin (HiCN) derivative can be conveniently and reproducibly prepared and is widely used for hemoglobin determinations. All forms of circulating hemoglobin are readily converted to HiCN except for sulfhemoglobin, which is rarely present in significant amounts. Cyanmethemoglobin can be measured accurately by its absorbance in a colorimeter.

Coulter Instrument Hemoglobin:

After the WBC solution is lysed, the system shines a beam of white light through the WBC aperture bath and then through an optical filter. This transmittance of light (525 nm wavelength) through a standard path length of Hgb solution is compared to the transmittance of such light in the same way through a reagent blank. The system converts this ratio to absorbance. It then converts absorbance to Hgb values in g/dL using a calibration factor.

Abott Cell-Dyne Hemoglobin:

This instrument uses a similar procedure, but is unique in that its reagent is cyanide-free.

Bayer Advia Hemoglobin:

Hemoglobin has dual readings - colorimetric or cyanmethemoglobin and corpuscular hemoglobin concentration mean.

Normal values:
Adult Male 14 - 17 g/dL
Adult Female 12.5 - 15 g/dL
Newborn 17 - 23 g/dL
3-month-old 9 - 14 g/dL
10-year-old 11.4 - 15.4 g/dL

Haematology Technologist

Automatic Cell Counter

Deal with the study of blood cells and blood diseases. Proficient in using automatic instruments for the assessment of:

* Haemoglobin
* White blood cell
* Platelet count

EDTA Blood Samples

Capable to detect various types of anaemia and other blood disorders like leukemia

Review on Thermo Fisher Scientific is the world's leader in serving science in Southeast Asia region

Thermo Fisher Scientific is the world's leader in serving science in Southeast Asia region

Thermo Fisher Scientific is the world's leader in serving science. Through the combined strength of its two premier brands, Thermo Electron and Fisher Scientific, Thermo Fisher Scientific positions itself as the largest provider of laboratory equipment and consumables, chemicals, bio-reagents, clinical and medical products, safety and clean room supplies, scientific instruments and software, and also laboratory furniture. With annual sales of more than $ 9 billion, we employ 30,000 people and serve over 350,000 customers within pharmaceutical and biotech companies, hospitals and clinical diagnostic labs, universities, research institutions, government agencies, as well as environmental and industrial process control settings.

With offices in both Malaysia and Singapore, we are able to extend our services and product offerings to many more customers in the Southeast Asia region. In Malaysia, Fisher Scientific (Part of Thermo Fisher Scientific) operates as the regional headquarters for South East Asia serving markets in Malaysia, Thailand, Vietnam and Philippines. In Singapore, Fisher Scientific (Part of Thermo Fisher Scientific) continues to serve our customers through the wide range of laboratory supplies and quality after-sales services.

Besides the vast product offering, Fisher Scientific (Part of Thermo Fisher Scientific) prides itself with its award-winning turnkey projects as well as its comprehensive technical services. Our distinctiveness lies in our ability to combine laboratory engineering to the laboratory equipment and testing methods. This extraordinary capability has given us the edge over our competitors in this region.

Review on XE- 5000 Full Automated Haematology Analyzer - Sysmex Product

Welcome to Sysmex Asia Pacific Pte Ltd

• Sysmex is a market leader in the field of clinical laboratory testing and health information technology products and services - supplying laboratories, hospitals and healthcare organizations worldwide.

XE-5000

• The XE-5000 is the latest fully automated hematology analyzer from Sysmex. Offering exceptional performance, XE-5000 further extends its functionality and capability beyond the routine.
• The XE-5000 incorporates advanced functionality for the measurement of immature cells, together with enabling the measurement of body fluids to extremely low concentrations of blood cells. The new system meets a wide range of customer needs from screening to specialized testing.

Features

• ‘Gold standard’ Fluorescence Flow Cytometry technology & special parameters as standard
• Unmatched performance and cost-effective reliability, employing a compact semiconductor laser unit as the heart of the flow cytometry system
• Rapid, dedicated body fluid analysis – including the differentiation of polymorphonuclear and mononuclear cells – without pre-treatment of the sample
• Comprehensive diagnostic decision-making, providing additional special parameters including Immature Granulocytes, Immature Platelet Fraction, Ret-He and Hematopoietic Progenitor Cells
• Switch from whole blood mode to body fluid mode at a simple push of a button
• Efficient automatic rinse and checking system for body fluid analysis, virtually eliminating sample to sample carryover
• Capacity for further enhancement by integration with other leading-edge Sysmex hematology technology – bringing totally automated analysis to the laboratory for rapid, accurate and cost effective results in a ‘walk-away’ environment
• Full network capability via the SYSMEX Network Communication System (SNCS), offering users Online QC, Remote Monitoring and Scientific Web Information services.

H5.m - Latest Hematology Lab Equipments

THE SEMIAUTOMATIC CELLCOUNTER, RELIABLE, ACCURATE AND COST EFFECTIVE

Single capillary analyzer, with impedentiometric reading principle, the H5.m determines the 7 basic hematologic parameters: RBC, WBC, Hgb, Hct, MCV, MCH and MCHC; photometric hemoglobin reading. Fast, accurate and easy to use, the instrument shows the data by the two displays and automatically prints them by the alphanumerical built-in printer. Equipped with a Self-Test program, the H5.m flags the presence of eventual functionality abnormalities by messages and suggestions for the problem solution. Numerical and progressive sample identification with direct possibility of insertion up to 3 digits by frontal panel. The RBC and WBC thresholds, for the numerical and volumetrical cell counting, for every kind of blood, are settable by the frontal panel. The system, economic and compact, can be used in any sort of laboratory, including, with very good results, the veterinary one.

Microlitre and Haematocrit Brushless Centrifuge

- Maximum speed : angle rotor: 14.000 rpm Haematocrit rotor: 12.000rpm

- Maximum RCF : angle rotor: 18.188 xg Haematocrit rotor: 14.810 xg

- Maximum capacity: Angle rotor: 24 x 2 ml. Haematocrit rotor: 24 x Capillary tubes

- Programmable microprocessor control system

- Pulse key

- Digital display for speed, time & RCF

- Timer: 1 - 99 min. and hold position

- Epoxy-polyester painted steel chamber

- Brushless induction motor

- Motor over heating protection

- Lid lock and interlock

- Delivered without rotor

GENIUS S - Hematology Lab Equipments

Automation in Hematology

The automatic cellcounter Genius S, with 40 µL of whole blood only and an analytical capacity of up to 65 samples/hour produces the following parameters: RBC, WBC, Hgb, Hct, MCV, MCH, MCHC, RDW, Plt, MPV, Pct, PDW percentage and absolute leukocyte screening values. The software is based on a complete system for data elaboration, is sophisticated but at the same time flexible and easy and immediate to use: the software’s wide menu give the operator prompt access to data management, file reports and quality control that provides Levey and Jennings Curves along with the Bull’s algorithm.


A set of flags, relative to all parameters, are given in case of pathological results including eosinophilies, monocytosis or any possible instrumental error. Moreover, for the samples with normal WBC differentiation the instrument detects automatically the 5 WBC subpopulations and a 3 WBC Part-diff discrimination for the pathological cases. For the determination of the hemoglobin Genius S uses a new lysing solution without cyanide. SEAC’s cleaning solution and self washing of blood sampling needle eliminates routine maintenance so that the service required from the operator is strictly reduced to a minimum.


Hemasampler The sample can be aspirated in completely automatic mode by the Hemasampler automated system, which is equipped with perforating needle for rubber tapped test tubes. Each rack has 24 positions. The rack rotation and the tube rocking mix the sample.

CLOT 2S - Hematology Lab Equipments

A NEW SYSTEM IN COAGULATION! FAST, ACCURATE AND EASY TO USE

The semiautomatic analyzer Clot 2S, developed for all the routine and urgency tests in coagulation, is the basic instrument for every kind of laboratory, ambulatory or hospital department; the Clot 2S is easy to use and at the same time it allows the programming of the methods, the standard curves elaboration and memorization; it is equipped with two independent reading channels, a 37°C thermostated plate, a liquid crystal display, a functional keyboard and a built-in printer for the results and the standard curve graphs print-out.

The counting system, starting with the addition of the starter reagent to the sample, automatically stops itself at the clot formation and the results are visualized and directly printed out on thermal paper. The results are expressed in seconds, or in Activity %, INR, RATIO or in Concentration according to the user selection and the measurement units are programmable between mg/dL, g/L, UI/mL. The alphanumerical keyboard allows the data insertion, the methods programming, the tests selection and the patient identification.


The incubation timers, for plasma and reagents, are independent and programmable for each method: this is an important aid for the correct working time respect. The Clot 2S is the only instrument, in his range, allowing the platelet aggregation test. The measurement is done using 0 and 100% automatic calibration with poor or rich platelet plasma and with the continuous mixing program activation: the trace is automatically printed out by the built-in printer with differentiated printing speed, programmable by software. The system can be connected to a Host computer by a RS232 serial port.

Cyanmethaemoglobin Method

Estimation of Haemoglobin Concentration can be done by using Cyanmethaemoglobin Method

Absorbance reading for Sample A = 0.410

Absorbance reading of Standard haemoglobin Solution = 0.336

The Standard Haemoglobin Concentration = 14.2 g/dL

• 20 µL (0.002 ml) of blood sample are used in this test. Blood is kept in EDTA tube.
• 6 ml of Drapkin reagent are used in this test
• Mixed 20 µL of blood to 6 ml ICSH (Drapkin) reagent in a test-tube.
• Test tube is kept in dark area for about 10 minutes
• Pour into a cuvette.
• Set the spectrometer to a wavelength of 540 nm. Measure the absorbance of the diluted test samples and reagent solution.
• Calculate the haemoglobin concentration. using the following formula
• Concentration Hb = O.D test/ O.D standard X concentration Standard

Calculation

Concentration of Hb = O.D test/O.D Standard x Conc. STD
= 0.410/0.366 X 14.2 g/dL
= 17.32 g/dL

Normal Range

• Adult Men = 14 – 17 g/dL
• Adult Female = 12.5 – 15 g/dL

Procedure for Leishman Staining and Differential WBC Count

Procedure of Leishman Staining

• Let thin blood film to air dry
• Then flood the slide with Leishman stain solution
• Wait for 8 minutes. Then flood off with distilled water in 2 – 3 seconds. Longer washing will remove stain
• Wait for 20 minutes. Observe the appearance of metallic cum that float on the surface of the slide.
• Then wash with tap water. Stand in a rack to drain and air dry.

Perform differential white blood cells count

• Using a low-power lens (10 x objectives) on the microscope, check the approximate differential cell distribution.
• Using the 40x objective high-power lens, perform a 200 cell differential count.
• 100x objective oil immersion lens should be used for examining fine intracellular details of the cells.
• Move the slide along the stage of the microscope in a broad battlement track running transversely across the body of the film, avoiding the edges completely.
• Record the numbers of each type of white cell. Calculate the percentage of each of the five basic leucocytes.

Normal Human Hemoglobins and their Variants

Hemoglobin A (also designated as A1) represents 98% of the normal adult hemoglobin. It consists of four subunits, two alpha (α) and two beta (β) chains to form a quaternary protein designated as a tetramer. This molecule is not an enzyme but can behave as one. Its major function is to combine with oxygen forming oxyhemoglobin. Each subunit binds with one oxygen molecule. When dissociated from oxygen, it is designated as reduced hemoglobin. Hemoglobin A2 and makes up to 3% of the normal adult hemoglobin. It is composed of two (α) and two delta (δ)chains. This hemoglobin is increased in β-thalassemia.

Hemoglobin F makes up to 80% of the hemoglobin in the newborn (at birth). It appears about the third month of fetal life and persists as the dominant hemoglobin until birth. It is composed of two (α) and two gamma (γ) chains. The gamma chains are beta-like. At six months to one year of age, the F type hemoglobin has essentially been replace by A1 hemoglobin. In the adult individual, it will make up <1%>

Hemoglobin Portland. This is the third embryonic hemoglobin that forms early in embryonic life and disappears in early fetal life. It is composed of two zeta (ζ) chains and two gamma (γ) chains. Hemoglobin Gower II. This is the second embryonic hemoglobin that forms prior to hemoglobin Portland and disappears in the third month of fetal life. It is the most important of the embryonic hemoglobins, making up as much as 60% of the three embryonic types. It is made up of two α-globulin chains and two ε (epsilon)-globulin chains. It is found in severe alpha thalassemia.

Hemoglobin Gower I is found during the first two months of embryonic life and only in small quantities. It is made up of two ζ-globulins and two ε-globulins.

Letters of the Greek Alphabet

* Α, α = alpha I, ι = iota Ρ, ρ = rho
* Β, β/ = beta Κ, κ = kappa Σ, σ/ς = sigma
* Γ, γ = gamma Λ, λ = lambda Τ,τ = tau
* Δ, δ = delta Μ, μ = mu Υ, υ = upsilon
* Ε, ε = epsilon Ν, v = nu Φ, φ = phi
* Ζ, ζ = zeta Ξ, ξ = xi Χ, χ = chi
* Η, η = eta Ο, o = omicron Ψ, ψ = psi
* Θ, θ = theta Π, π, = pi Ω, ω = omega

This alphabet is provided for your information.


Other hemoglobin variants found in the normal adult are:

*Hemoglobin AIc which may make as much as 3.5% of the total adult hemoglobin. It is found in concentrations up to 30% (range = 8 to 30%) in diabetics. This is hemoglobin A1 that has undergone non-enzymatic glycosylation (with glucose via the Schiff reaction).
*Hemoglobin AIa is a hemoglobin A1 that is glycosylated with either fructose-1,6 phosphate molecule (AIa1) or a glucose-6-phosphate molecule (AIa2 ).

More about the glycosylated hemoglobin variant

Hgb A1, when bound to glucose, is called glycosylated hemoglobin. This phenomenon is dependent upon containing valine as a N–terminal amino acid on the beta chain. Glucose, if it makes contact with this terminal amino acid, will spontaneously bond in a non-enzymatic reaction to valine and form an aldimine or Schiff’s base. In this stage, this new intermediate form of hemoglobin is designated as “pre- Hgb AIc” and is somewhat unstable. This Schiff base will undergo a slow transformation (called the Amadori rearrangement) to form a ketoamine, a stable form of Hgb AIc. This process will occur throughout the life of the RBC.

The rate of Schiff base formation in the blood is blood glucose concentration dependent. In the normal individual, up to 8% of the hemoglobin (N: 5% to 8%) will be in the different forms of glycosylated hemoglobin (Hgb AIa, Hgb AIb, and Hgb AIc). An individual whose diabetes is under control will have an expected range of 8% to 15% glycosylated hemoglobin where the out-of-control diabetic may have a range of 15% to 30%.