Erythrocyte Sedimentation Rate: Westergren Method

Erythrocyte
Sedimentation Rate
Westergren Method
• The erythrocyte sedimentation rate (ESR) is a common hematological test
for nonspecific detection of inflammation that may be caused by infection,
some cancers and certain autoimmune diseases.
• It can be defined as the rate at which Red Blood Cells (RBCs) sediment in a
period of one hour.
Three Stages of Mechanism
▪ Stage of aggregation : It is the initial stage in which piling up of RBCs takes place. The
phenomenon is known as Rouleaux formation. It occurs in the first 10-15 minutes.
▪ Stage of sedimentation : It is the stage of actual falling of RBCs in which sedimentation

occurs at constant rate. This occurs in 30-40 minutes out of 1 hour, depending upon the
length of the tube used.
▪ Stage of packing : This is the final stage and is also known as stationary phase. In this,
there is a slower rate of falling during which packing of sedimented RBCs in column occurs
due to overcrowding. It occurs in final 10 minutes in 1 hour.
• Two principal measures:
• Westergren Method
• Wintrobe Method
• Westergren method is recommended by CLSI

Principle
• A measure of the degree of settling of erythrocytes in plasma in an
anticoagulated whole-blood specimen during a specified period of time
• ESR is directly proportional to red cell mass and inversely proportional to

plasma viscosity
Specimen Requirements
• Whole blood anticoagulated with EDTA (modified Westergren Method)
• Actual collection must not be more than 30 seconds
• 3.8% sodium citrate (original Westergren Method) is also used in some

laboratories
• Blood kept at room temperature must be set up within 2 hours

• Storage at 4OC is stable for 6 hours – 12 hours
Westergren ESR Tube
• Made of thick-walled glass or hard plastic with an overall length of 300mm
(±0.5mm), length of blood column of 200mm, external diameter of 5.5mm
(±0.5mm), tube bore of 2.65mm (±0.15mm)
• Markings on the tube should extend over the lower 200mm of the pipet
• Numbered from “0” at the top of the scale to at least “180” at the lower end of the
pipet, with increments of 10mm or less
• The rack to hold the tubes must not allow leakage from the pipets and must be held
motionless in a vertical position
Westergren Tube set-up
Procedure
• The original and modified Westergren methods differ only in the
anticoagulant used and the time at which the blood is diluted
#1 (Specimen Preparation)
• Original Westergren
• Whole blood is drawn into 3.8% sodium citrate
• No further dilution is performed
• Modified Westergren
• Combine 2.0mL of well mixed EDTA-anticoagulated blood with 0.5mL of 0.85%
NaCl or 0.5mL 3.8% sodium citrate
• Most preferred by laboratories (EDTA is readily available)
#2
• Place blood containers in pipet rack directly
under each hole.
• Insert a Westergren pipet through each hole
into the blood mixture.
#3
• Firmly press a safety bulb or hand pump on
top of the Westergren pipet and watch the
blood being aspirated into the “0” mark.
• If using a plugged pipet, the plug should be

well soaked with blood.
• Repeat for all specimens to be tested.

#4
• After exactly 60 minutes, record the distance, in millimeters, from the bottom
of plasma meniscus to the top of the column of sedimented erythrocytes.
• Do not include the buffy coat in the reading.

#5
• Report results as “ESR = ____mm”
• Note that this test does not measure a rate, rather, it measures distance after
a specified time
• Dispose of all Westergren tubes carefully (hazardous)

Reference Range (in 1 hour)
Gender Age Group Reference Value
Males <50 years old 0 – 15mm
>50 years old 0 – 20mm
Females <50 years old 0 – 20mm
>50 years old 0 – 30mm
Children 0 – 10mm
Comments and Sources of
Error
• ESR is considered a nonspecific indicator of tissue damage, and it is increased in a
number of disorders
• Conditions in which rouleaux formation is inhibited (SC anemia, Hb CC, and

spherocytosis) may be accompanied by normal sedimentation rates
• Leaving the specimen for more than 2 hours at room temperature will cause
erythrocytes to become spherical and inhibit rouleaux formation
• Over-anticoagulation causes lower values
• Slight tilting of the tube increases ESR values
• Marked changes in temperature affects ESR results

• High temperature = false increase
• Low temperature = false low
• Presence of anemia invalidates the ESR as a tool to detect rouleaux

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Erythrocyte

▪ Stage of sedimentation : It is the stage of actual falling of RBCs in which sedimentation

• Westergren method is recommended by CLSI

• ESR is directly proportional to red cell mass and inversely proportional to

• 3.8% sodium citrate (original Westergren Method) is also used in some

• Blood kept at room temperature must be set up within 2 hours

• If using a plugged pipet, the plug should be

• Repeat for all specimens to be tested.

• Do not include the buffy coat in the reading.

• Dispose of all Westergren tubes carefully (hazardous)

• Conditions in which rouleaux formation is inhibited (SC anemia, Hb CC, and

• Slight tilting of the tube increases ESR values

• Marked changes in temperature affects ESR results

• Presence of anemia invalidates the ESR as a tool to detect rouleaux

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