INTRODUCTION TO

HEMATOLOGY
Jardine Jade Albert C. Valdez, RMT
Capitol Medical center College
Hematology

 The study of blood.

 BLOOD is the circulatory fluid of the
cardiovascular system which is circulating
constantly through a closed system circuit
of tubes.
Liquid Portion

 Plasma – Straw and pale yellow liquid portion

of blood after coagulation has been prevented.
 Serum – Clear and yellow liquid portion of
blood after separation of the clot.
Composition of Plasma or Serum

■ A. Chemicals
• Sugars
• Proteins
• NPN (non protein nitrogenous subs)
• Enzymes and hormones
• Mineral and electrolytes
• Other chemical substances
■ B. H2O (water)
■ C. Food materials/ nutrients
Solid Portion

 Erythrocytes/ RBC
 Leukocytes/ WBC
 Thrombocytes/ Platelets
1. Red blood cells – with hemoglobin, carrier of oxygen
and carbon dioxide.
– Other names are:
– Erythrocytes
– Erythroplastids
– Normocytes
– Akaryocytes (without nucleus)
– Discocytes
2. White Blood cells – Responsible for phagocytosis and
produces Antibodies.
– Other names are:
– Leukocytes
– Leukoplastids
– Types:
– Granular
– Agranular
Color reaction of granules

 Neutrophils – lilac pink

 Eosinophils – red to orange
 Basophils – dark blue to black
■ 3. Platelets – other names are: thrombocytes,
thromboplastids
Functions of platelets:
 Hemostasis
 Blood coagulation
 Clot retraction
 Gaseous portion

Oxygen
Carbon dioxide
Characteristics of blood

1. Volume: 7-8% of the body weight

2. Color: Dark red – low oxygen weight; bright
scarlet red – high oxygen content
3. Viscosity: 5 times more viscous than water
4. Specific gravity: 1.055 – 1.065
5. Reaction: 7.35-7.45
 In fluid form it coagulates within 5-10
mins in vitro.
 Heparin is natural anticoagulants.
 Red in color due to hemoglobin content
 Hemoglobin is the main component of
RBC which is responsible for the oxygen
transport of the blood.
Function of Blood.

■I. Metabolic
■II. Defensive function
Metabolic function

 Respiration
 Nutrition
 Excretion
 Regulation of water balance
 Regulation of body temperature
 Transport of hormones
 Maintenance of acid base balance in the body
Defensive function

Production of immune globulins

Functions as phagocytes:
Neutrophils
Eosinophils
Basophils
Monocytes
Blood that produces muramidase

Neutrophils – 1st line of defense

Monocytes – 2nd line of defense
Lymphocytes - traces
Hemopoiesis

■ Manufacture and development of blood cells

– Erythropoiesis – RBC
– Leucopoiesis – WBC
– Thrombopoiesis – Platelets
■ Hemocytoblast
■ Hemohistioblast
■ Hemopoietic reticulum cell
3 stages of hemopoiesis

1.Mesoblastic stage (yolk sac)

2.Hepatic
3.Myeloid or medullary
Mesoblastic stage (yolk sac)

■ 2 weeks fetal life – formation of blood island

■ 9 weeks – Primitive erythroblast (PE)
– Primitive erythroblast produces hemoglobin for
oxygen needs of fetus.
– Embryonic hemoglobin: Portland, gower- 1 and
gower- 2
Hepatic

■ 2nd month – 3rd month

■ Liver – the chief site of hemopoiesis
■ Increase in more definite erythroblasts,
granulocytes, monocytes, lymphocytes and
megakaryocytes (mother of platelets)
■ Hemoglobin A, A2 and F
Myeloid or Medullary (Red BM)

■ 7th month
■ Start in red bone marrow
■ Continues in other organs
■ Extramedullary stage (a few weeks before birth) 
adult life
■ Red bone marrow
■ Medullary hemopoeisis
Stromal

■ Skull
■ Sternum Flat bones:
■ Pelvis Active adult hematopoiesis
tissue (colony forming
■ Ribs
units in red bone marrow)
■ Vertebrae
Blood volume:

■ Normal male adult: 5.8 L (5,800 ml)

■ Normal female adult: 4.5 L (4,500 ml)
Terms:

a. Normovolemia
b. Hypovolemia
c. Hypervolemia
d. oligemia
Decrease blood volume in the following:

1.Loss of whole blood (hemorrhage)

2.Loss of RBC (hemolytic anemia)
3.Loss of Plasma (severe burns)
4.Loss of body H2O (sweating)
Increase blood volume in the following:

1.During IV injection of fluids

2.Blood transfusion
3.Excessive fluid intake
Normal blood volume:

■ Plasma volume – 55% of total blood volume

■ Packed cell volume – 45%

100%
Anticoagulants:

■ EDTA
■ Citrates
■ Heparin
■ Double oxalates
■ Fluorides
■ Defibrinated blood
■ Use of siliconized glasswares
EDTA

■ The disodium or tripotassium salt of

ethylenediaminetetraacetic acid is the most
commonly used anticoagulant in hematology.
■ Blood coagulation is inhibited by chelating or binding
of calcium ions.
■ Optimal concentration is 1.5 mg/ml of blood.
Advantage of EDTA

■ Best for smears (preserves cellular

morphology)
■ Best for platelet count and WBC count
■ Prevents platelets aggregation
Disadvantage of EDTA

■ Cell shrinkage (HCT and ESR are

decreased)
■ Degenerative white cell changes
■ Swelling and breaking up of platelets
Citrate

■ Inhibits blood clotting by binding calcium in a

soluble complex and is used for many
coagulation studies
■ Trisodium citrate is the choice today for many
coagulation studies since it preserve labile
factors V and VII
■ Most satisfactory for platelet aggregation
studies
Disadvantage of citrate

■ If HCT exceeds 50amount unbound citrate

in the citrate plasma mixture causes
prolonged clotting times
Heparin

■ Coagulation is inhibited by its interaction with

antithrombin III and subsequent inhibition of
thrombin
■ Concentration is 15-30 units/ml of blood
■ Has many anticoagulant actions
■ Cause aggregation of cells
Double oxalates

■ 3:2
■ Preparation:
– Ammonium oxalate – 1.2 grams
– Potassium oxalate – 0.8 grams
– Neutral formaldehyde - 1.0 ml
– Dist water – 100 ml
Fluorides

■ By forming weakly dissociated calcium components

Specimen Collection (blood collection)

■ Phlebotomist – person trained in blood

collection, provides link top the patient and
laboratory.
■ Good interpersonal skills are necessary
■ Must ensure confidentiality
Types of blood collection

1. Skin puncture
2. Venipuncture
3. Arterial puncture
Skin puncture

■ Sites of choice:
– Earlobe
– Palmar surface of finger (3rd and 4th)
– Plantar surface of heel and big toe
Blood collected by skin puncture:

■ Peripheral blood
■ Capillary blood
■ Arteriolar blood
Skin puncturing/microsampling – mixture of
capilliary, venous and arterial blood and
contains interstitial and intrecellular fluids
Uses of skin puncture

1. Infants
2. Safer means of blood collection
3. In case of obesity, burns and severely small
and damaged veins
4. IV fluid is flowing to the accessible vein
Sites to Avoid

1. Inflammed and pallor areas

2. Cold and cyanotic
Increased cell
count and WBC
3. Congested and edematous
4. Scarred and heavily calloused
Advantage of skin puncture

■ Easily accessible to operator

■ Easily to manipulate
■ Ideal for peripheral blood smears
■ Less intimidating
Disadvantage of skin puncture:

■ Small volume of blood are collected

■ RBC count, hematocrit and hemoglobin,
platelets count are lower
■ WBC count are higher
Sources of errors (skin puncture)

■ Hemolyzed specimen
■ Failure to dry the site
■ Excessively deep skin puncture
■ Failure to wipe off the first drop of blood
■ Milking or area
■ Accidental capturing of bubbles from capillary tube
Procedure of skin puncture

1. Disinfect area of choice

2. Puncture to a depth of 3 mm
3. Wipe off 1st drop of blood
4. Use new drop of blood/test
5. Press punctured area with cotton
Sequence of blood collection for CBC:

1. Blood smear
2. Hemoglobin
3. Hematocrit
4. WBC count
5. RBC count
Venipuncture

■ Blood collection method of venipuncture:

a. Syringe method
b. Evacuated method
c. Butterfly method
Sites of choice for venipuncture:

■ Newborn to 18 mons old baby

1. External jugular veins
2. Temporal veins
3. Superior longitudinal sinus
■ 18 months to 3 years old
4. Femoral vein
5. Long saphenous veins
6. Popliteal vein
7. Ankle vein
■ 3 years to adult
8. Wrist vein
9. Veins on dorsal of hand/ finger
10.Veins on antecubital fossa – most ideal site
■ Median basilic vein
■ Median cephalic vein
■ Median vein
Procedure in venipuncture:

1. Apply tourniquet
2. Palpate the veins
3. Disinfect the area (circular motion)
4. Enter needle into the vein (bevel up in swift motion)
5. Release the tourniquet
6. Obtain blood
7. Prepare dry sterile cotton over site of puncture
8. Withdraw needle from the vein
9. Press dry cotton over the punctured area
10.Apply pressure directly on punctured area while
patient holds his arm up high for at least 3 minutes.
(do not flex the patient elbows)
11.Remove needle from the adapter or syringe and
dispose blood gently into suitable container.
Sequence:

a. Sterile tubes
b. Tubes without additives
c. Coagulation tubes (PT)
d. Tubes with other additives
Advantage of ETS over the syringe
method:
1. For collecting and storing blood
2. With built in anti-coagulants
3. Disposable
4. Multiple drawing of blood possible
5. Less exposure of blood to contaminants
Disadvantage of the syringe

1. Difficulty in drawing blood

2. Can not collect full 10 cc of blood only 7-8 cc as
plunger may give way
3. Multiple specimen collection will require several
venipuncture more traumatic to the patient
4. Easy coagulation of blood in the syringe
5. Need to transfer specimen from syringe to test tube
6. Messy, time consuming may take an average of 10
mins.
7. Need to add appropriate anticoagulant into test
tube
8. Results may be questionable
9. Increase risk of contamination due to possible
contact of specimen with the medical technologist
Complications of venipuncture:

I. Immediate local complications

II. Delayed local complications
III.Delayed general complications
Immediate local complications:

1. Hemoconcentration – aggregation of blood cells in the site of

puncture.
2. Failure of blood to enter the syringe:
1. Collapsed vein
2. Hitting the vein thru and thru
3. Hitting just the wall of the vein
1. Sclerotic (thick wall)
2. Movable vein
3. Circulatory failure
– Nervousness
– Coma/shock
4. Faiting or syncope
– Nervousness
– Shock/coma
Delayed local complication

1. Hematoma
2. Thrombosis of the vein – formation of clot in the
vein at the site of puncture
3. Thrombophlebitis – inflammation of vein at the site
of puncture
Delayed general complication

1. Serum hepatitis – HBV

– Infection are in the blood and by products and also in
body fluids.
2. AIDS – caused by HIV
– Through blood and its by product
– Sexual transmission
– Placental transmission
– Contaminated needles and surgical instruments
End of lecture!!!!