Erythrocyte Sedimentation Rate

• Erythrocyte sedimentation rate (ESR) is the rate at which the

erythrocytes settle down.
• Normally, the red blood cells (RBCs) remain suspended uniformly in
circulation. This is called suspension stability of RBCs.
• If blood is mixed with an anticoagulant and allowed to stand on a
vertical tube, the red cells settle down due to gravity with a
supernatant layer of clear plasma.
• ESR is also called sedimentation rate, sedrate or Biernacki reaction.
It was first demonstrated by Edmund Biernacki in 1897
DETERMINATION OF ESR
There are two methods to determine ESR
1. Westergren method
2. Wintrobe method
WESTERGREN METHOD
In this method, Westergren tube is used to determine ESR
Westergren Tube
The tube is 300 mm long and opened on both ends
 It is marked 0 to 200 mm from above downwards.
 Westergren tube is used only for determining ESR. 1.6 mL of blood is mixed with
0.4 mL of 3.8% sodium citrate (anticoagulant) and loaded in the Westergren tube.
The ratio of blood and anticoagulant is 4:1.
The tube is fitted to the stand vertically and left undisturbed.
The reading is taken at the end of 1 hour
WINTROBE METHOD
• In this method, Wintrobe tube is used to determine ESR.
• Wintrobe Tube Wintrobe tube is a short tube opened on only one end
• It is 110 mm long with 3 mm bore.
• Wintrobe tube is used for determining ESR and PCV.
• It is marked on both sides. On one side the marking is from 0 to 100
(for ESR) and on other side from 100 to 0 (for PCV).
• About 1 mL of blood is mixed with anticoagulant,
ethylenediaminetetraacetic acid (EDTA).
• The blood is loaded in the tube up to ‘0’ mark and the tube is placed
on the Wintrobe stand
• the reading is taken after 1 hour
NORMAL VALUES OF ESR
By Westergren Method
• In males : 3 to 7 mm in 1 hour
• In females : 5 to 9 mm in 1 -hour Infants : 0 to 2 mm in 1 hour
By Wintrobe Method
• In males : 0 to 9 mm in 1 hour
• In females : 0 to 15 mm in 1 hour
• Infants : 0 to 5 mm in 1 hour
SIGNIFICANCE OF DETERMINING
ESR
• Determination of ESR is especially helpful in assessing the progress of
patients treated for certain chronic inflammatory disorders such as:
1. Pulmonary tuberculosis
2. Rheumatoid arthritis
3. Polymyalgia rheumatica (inflammatory disease characterized by pain
in shoulder and hip)
4. Temporal arteritis (inflammation of arteries of head).
VARIATIONS OF ESR
• PHYSIOLOGICAL VARIATION
1. Age: ESR is less in children and infants because of more number of
RBCs.
2. Sex: It is more in females than in males because of less number of
RBCs.
3. Menstruation: The ESR increases during menstruation because of
loss of blood and RBCs
4. Pregnancy: From 3rd month to parturition, ESR increases up to 35
mm in 1 hour because of hemodilution
PATHOLOGICAL VARIATION
• ESR increases in diseases such as the following conditions:
1. Tuberculosis
2. All types of anemia except sickle cell anemia
3. Malignant tumors
4. Rheumatoid arthritis
5. Rheumatic fever
6. Liver diseases.
• ESR decreases in the following conditions:
1. Allergic conditions
2. Sickle cell anemia
3. Peptone shock
4. Polycythemia
5. Severe leukocytosis.
FACTORS AFFECTING ESR
• FACTORS INCREASEING ESR
1. Specific Gravity of RBC When the specific gravity of the RBC
increases, the cells become heavier and sedimentation is fast. So
ESR increases
2. Rouleaux Formation Rouleaux formation increases the ESR. Globulin
and fibrinogen accelerate the rouleaux formation
3. Increase in Size of RBC When the size of RBC increases (macrocyte),
ESR also increases.
FACTORS DECREASING ESR
1. Viscosity of Blood Viscosity offers more resistance for settling of
RBCs. So when the viscosity of blood increases, the ESR decreases
2. RBC count When RBC count increases, the viscosity of blood is
increased and ESR decreases. And when the RBC count decreases, ESR
increases
Determination of the Pack Cell Volume
• Hematocrit (HCT) Also known as the hematocrit test, the PCV or Packed Cell
Volume Test is a test done to diagnose polycythemia, dehydration or anemia in certain
patients.
• The hematocrit, also known by several other names, is the volume percentage of red
blood cells in blood, measured as part of a blood test. The measurement depends on
the number and size of red blood cells. It is normally 40.7–50.3% for males and
36.1–44.3% for females.
• The packed cell volume (PCV) is a measurement of the proportion of blood that is
made up of cells. The value is expressed as a percentage or fraction of cells in blood.
• It is the volume of RBCs packed at the bottom of a hematocrit tube when the blood is
centrifuged. It is also called hematocrit value or erythrocyte volume fraction
(EVF).
SIGNIFICANCE OF DETERMINING
PCV Determination of PCV helps in
1. Diagnosis and treatment of anemia
2. Diagnosis and treatment of polycythemia
3. Determination of extent of dehydration and recovery from
dehydration after treatment
4. Decision of blood transfusion
VARIATIONS IN PCV
• INCREASE IN PCV
1. Polycythemia
2. Dehydration
3. Dengue shock syndrome: Dengue fever (tropical disease caused by flavivirus
transmitted by mosquito Aedes aegypti) of grade III or IV severity
• DECREASE IN PCV
1. Anemia 2. Cirrhosis of liver 3. Pregnancy
4. Hemorrhage due to ectopic pregnancy (pregnancy due to implantation of
fertilized ovum in tissues other than uterine wall), which is characterized by vaginal
bleeding.
METHOD OF DETERMINATION
Blood is a mixture of cells and plasma. The packed cell volume (PCV) is a
measurement of the proportion of blood that is made up of cells.
• Roughly estimates it is three times of the hemoglobin value.
Methods
1) Macro method ( wintrobe )
2) Micro method
Wintrobe Method
Principle
• When anticoagulant blood is centrifuged in wintrobe tube at high speed 300rpm
the erythrocytes sediments at bottom.
• The red cell column is called pack cell volume.
• PCV represents the volume of the cells per unit volume of the whole blood and
expressed as a percentage.
Wintrobe Method
• Open at top but close at lower end
• Internal diameter 2.5mm
• 11 cm long
• 10cm (100 mm) marking long
• Total diameter 3mm.
• Each 1cm gap is divided into 10 millimeter
• From above to downward (for ESR)
• From below to up (for PCV)
Procedure
Collect the blood in EDTA vial.
Fill the wintrobe tube up to 10 mark of the collected blood by Pasteur pipette.
Now place the tube in centrifuge at 3000 rpm for 30 minutes
Now note upper marking as percentage.
Microhematocrit
• Ideal for finger prick
• Useful in pediatric patients

llbbyybyy length 7.5cm, internal diameter 1mm

Procedure
Take fresh drop of blood.
Touch the red circle end of the heparinized capillary tube to drop.
Hold the tube in horizontal position and allow the blood to enter
in tube in one half to three full.
Seal one end of the tube.
Place the capillary tube in the microhematocrit with plug end and
away from center.
Centrifuge for 4 minutes
Measure the height of the red cell column and height of the cells
plus the plasma.
Hct = height of red cells/ height of the red cells and plasma (mm)
X 100
Normal Range %
Dog 37-55 sheep 27-45

Cat 30-45 Goat 22-38

cow 24-46 llama 29-39

Horse 32-48 Ostrich 32

Human 36-49 Chicken 25-40