7. ERYTHROCYTESEDIMENTATION RATE (E.S.R.

)
Aim-Estimation of erythocytessedimentation rate in mm ofyour own blood sample
Definition:The rate at whicherythrocytes settle down under standard conditions.
Methods: [Link] [Link]
Westergren method
Apparatus:
1. Westergren tube. It is a straight glass tube 300 mm in length and 2.5 mm diameter. A scale
graduated in mm. extends overlower 200 mm.
2. Rack : Specially made racks with adjustable levelling screws are available for holding the
Westergren tube firmly in an exactly verticalposition.
Reagents: Sodium citrate 3.8%. Procedure
1. With usual aseptic precautions collect the venous blood. Use 3.8%
sodium citrate as an
anticoagulant (4 volumes ofblood: 1 vol. of3.8% citrate).
Then draw the blood into the Westergren tubeupto zero
2.
mark by sucking
3. Place thelube exactly vertical in the rack and keep It undisturbed
for onc hour.
4. At the end of one hour read the height of clear plasma above
the upper limit of column of
sedimenting cells to the nearest mm. Thisindicates E.S.R.
Normal range: maies 0-5 mm at the end of one hour
females 0-7 mm àt the end of one hour
Wintrobe method
Apparatus:
1 Wintrobe tube: this is a glass tube approximately 3 mm in
in length calibrated at 1 mm intervals to 100 internal diameter and about110 mm
mm.
Reagent; Ammonium potassiumoxalate. (3:2)
Procedure:
1 With usual aseptic precautions collect the venous
blood.
2 Use2 mg of oxalate mixture/ml. ofblood as an
3 Using long nozzle dropper (reaching upto the
anticoagulant.
mark. bottom of tube), pillthe Wintrobc's tubcupto zero
4
Avoid airbubblesandfrothing of blood.
5 Fix the tube vertically in the stand
without disturbing for 1 hour.
6. At the end of 1 hour read the
height of clear plasma above the upper
sedimenting cells to thenearest [Link] indicates E.S.R.
limit of column of
Normal range: Males -0-9 mm atthe end on one hour.
Females -0 - 20 mm at the end of onchour.
Factors Affecting ESR
1. Rouleanx formation. The adhesion and
piling red cells one upon the other
coins)vcafled rouleaux formation. Inthe circulating (like a pile of
their constant movement and due the RBCs remain separate from
to cach other to
clectrostaticcharges. However, they tendtheir mutual repulsion rcsulting from thcir
to stick to each othcr and form negative
ablood film orin a test tube. roulcaux and clumps in
In health the rouleaux are small
and settle slowly. Ilowever, under
formation increases and thus it is the main certain conditions, roulcaux
and rate of roulcaux formation is as follows. (actor that increascs the ESR. The importance of the sizc

18
 The surface area of a single red cell is about 150p m'. and 10 red cells would have a surlace arca
of 1500p m' -when theydo not form apile. But when these 10 redcells form a rouleaux (pile), their
surface area is only 600 ur m'while their mass has increased 10 tines. Thus, when the red cells form
rouleaux, there is a greater increase in the masses of these red cells as compared to increase in their
surface area. This decreases the retardation causcd by plasma viscosity, which, in turn, ncreases the
rate of settling of red cells, and thus the ESR.
Factors affecting rouleaux formation, and thus, ESR are:
a. Concentration of large, asymmetric molecules of fibrinogen and globulins. An increase in the
concentration of these plasma proteins promotes rouleaux formation (and hence ESR); this is
seen in almost all infections (acute andchronic, tissue destruction due to any cause, andwasting
diseases.
b. Bacterial proteins and toxins, and products of inflammation and tissuc destruction. Absorption
of these materials into the blood, and changes in plasma protein pattern probably promotes
rouleaux by coating the red cells thus increasing their stickiness and/or altering their surface
charges.
C. Number of red cells (Ratio of RBCs to plasma; normal * 45/55). A decrease In RBC count promotes
rouleaux formationand increases the ESR while an increase in redcells decreases ESR.
d Size and shape of redcells. An increase in MCV,adecrease in MCH,and spherocytosistend to slow
down rouleaux formation and hence ESR.
2 Viscosity of blood. Increased viscosity decreases ESR, while decreased viscosity has the opposite
effect. (Viscosity ofwater plasma blood= 1:3:5).
the red
3 anticoagulants used. Use of (fluid anticoagulants (eg, 3.8% sodium citrate) changes
cell/plasma ratio, which increases the ESR

QUESTIONS:
1
What are the physiological and pathologicalvariations of E.S. R.?
2 What is the clinicalsignificance of E.S.R.?
method?
3
What-are the advantages and disadvantages of theWintrobe & Westergren
4 Draw labelled draw.
• PHYSIOLO GCAL
1- PtaOLOiceu variaone
in
-
neuwborn becauce KBC coUnt is m ore.
0) Age: louesr vawe
b) Se M:igner
) €SRt as fibrinogenT dring preqnaneyIhis
Prequawy: increac e y 00teaun qorMaon.
a) Jeuperart: Et ESK« Venper aMYe
Hain pysioto
PATHO Lo6IeAL variaons:
phrosic
cytaea, euke nias, aiquaM cis, ne exe.
ahauia, po
ing a di's ease.
vaues aYe o uo Ie ju oi a qnos
2:
ES R
proqnosác cIe
v 4 ÉSR ↑ ir ind carns Speed o
3 taas ony 19
3Ome disease iu lbody
 (rom hronie ilness su ch
Jhe progres s puá nt sufering9
as TB, rhumaroid
aYHhriás 'ere.
DIAGRAMS:

→PLASMA
wh
blood
100 N

RubbN Oorke
)