This document discusses erythrocyte sedimentation rate (ESR), packed cell volume (PCV), and red blood cell indices. It defines these tests and provides the normal ranges. ESR is measured using the Westergren or Wintrobe method and measures how fast red blood cells fall in a tube. PCV measures the ratio of red blood cells to whole blood. Red blood cell indices include mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration which provide information about the size and hemoglobin content of red blood cells. The document discusses factors that affect the tests and their clinical significance.

1. ESR, PCV AND BLOOD
INDICES
DR. JANANI MATHIALAGAN
1ST
YEAR POSTGRADUATE
PATHOLOGY

2. OBJECTIVES OF THIS
CLASS
To perform and interpret ESR
To perform and interpret PCV
To perform and interpret Blood Indices

3. ERYTHROCYTE SEDIMENTATION
RATE (ESR)
ESR is the measurement of the rate of sedimentation of red cells
in anti-coagulated blood.
Blood is allowed to stand for 1 hr in an open-ended glass tube
mounted vertically on a stand
Length of column of plasma above the red cells is measured in
mm.

4. Anticoagulated blood is drawn up into a tube of standardized
dimensions and left in a vertical position for exactly one hour
By that time, the red cells would have separated and settled
from the plasma.
Upper plasma column is recorded by reading from the scale
on the side of the tube.
Measures the distance that RBCs will fall in a vertical tube
over a given time period
Initial screening tool and also as a follow-up test – monitor
therapy and progression or remission of disease

5. Three definite phases:
• First or Lag Phase (10mins) – red cells form a characteristic
rouleaux pattern (aggregation) and sedimentation is generally
slow. (Pack of coins)
• Decantation Phase (40mins) – The rate accelerates in this phase;
fast settling or sinking of RBCs
• Final Packing Phase (last 10mins) – slows again as red cell
aggregates pile up at the base of the tube. There is slow
sedimentation.

6. FACTORS AFFECTING
ESR
1. Plasma factor
2. RBC factor
3. Technical factor

7. PLASMA FACTORS
Increased fibrinogen increases rouleaux formation
thereby increasing ESR
S. haptoglobulin , C - reactive protein & cholesterol
also increases ESR
Albumin and lecithin decreases sedimentation i.e.
decreasing ESR

8. RBC FACTORS
Primarily through changes in number and/or shape
Anemia responsible for increased ESR
◦Microcytes – sediment more slowly
◦Macrocytes – sediment faster

9. Poikilocytosis retards ESR because abnormal shape
hampers rouleaux formation
Anticoagulants – Sodium citrate & EDTA doesnot affect ESR
oxalates & heparin may affect

10. TECHNICAL FACTORS
Poor temperature control
Length and bore of the tube
Vibration
Verticality

11. METHODS
Westergren’s method
Wintrobe’s method
Micro - ESR
Automated systems
Zeta sedimentation

12. The method for measuring the ESR recommended
by the International Council for Standardization in
Haematology (ICSH)
Based on that of Westergren, who developed the
test in 1921 for studying patients with pulmonary
tuberculosis.

13. CONVENTIONAL WESTERGREN
METHOD
The recommended tube is a straight
glass or rigid transparent plastic tube
30 cm in length
2.55 mm in diameter.
Bore must be uniform
A scale graduated in mm extends
over the lower 20 cm.

14. For the diluent, 3.8 g/dl Trisodium citrate used
Dilution – 1:4
0.25ml trisodium citrate : 1ml blood
Mix the blood sample thoroughly and then draw it up into
the Westergren tube to the 200 mm mark by means of a
rubber teat or a mechanical device

15. Place the tube exactly vertical
and leave undisturbed for
exactly 60 min, free from
vibrations and draughts and
not exposed to direct
sunlight.
Then read to the nearest 1
mm the height of the clear
plasma above the upper limit
of the column of sedimenting
cells.
Westergren pipette filled with blood
and placed vertically on the rubber
cork in the rack

16. ERYTHROCYTE SEDIMENTATION
RATE
Average ESR value by Westergren Method:
Male – 3-5mm
Female – 4-7mm

17. PROCEDURE – WINTROBE METHOD
1. Add well mixed double oxalate / EDTA blood to the zero mark of
the Wintrobe tube, using a pipette
Avoid air bubbles
2. Place in vertical position in a rack and let sit for 60 minutes
3. Read and record results in millimeter (distance which the cells
have settled)

18. Average ESR value by Wintrobe’s Method:
◦Males: 0 – 9mm/hr
◦Females: 0 – 20mm/hr
◦Children: 0 – 13mm/hr

19. Increased ESR
•Chronic infections e.g. Tuberculosis
•Extensive/ Chronic inflammation
•Collagen vascular disorders
o Systemic Lupus Erythromatosus
o Rheumatoid asthritis
o Systemic Sclerosis
•Shock
•Active syphilis
•Active infectious infections

20. Decreased ESR
•Newborns
•Congestive heart failure
•Polycythemia
•Marked leukocytosis
•Allergic states
•Sickle cell anemia

21. Ratio of volume of RBCs to that of whole blood
It indicates relative proportion of red cells to plasma
Expressed in percentage.
Also called hematocrit or erythrocyte volume fraction
PACKED CELL VOLUME

22. Methods:
1. Macrohematocrit method (Wintrobe Method)
2. Microhematocrit method
3. Electronic Method

23. WINTROBES METHOD
•Wintrobe’s tube – 110mm long,
internal bore 2.5mm &
a flat inner base.
Graded 0-10 on both sides.

25. Method:
1.Mix the anticoagulant blood sample thoroughly
2.Draw blood in a Pasteur pipette
3.Fill the tube upto 10 mark
4.Centrifuge the sample at 2000-2300 rpm for
30mins
5.Take the reading of the length of the column of
red cells

27. Buffy coat- WBC&
PLATELETS.
UPPER MOST LAYER – PLASMA
•Yellowish-Jaundice
•Pink-haemolysis
•Milky-hyperlipidemia

29. PCV reading

30. PRECAUTIONS
 Use recommended amount of EDTA
 Test done with in 6-8 hours
 Wintrobe tube should be filled from below upwards
so that no air bubble is trapped.

31. INCREASED PCV
Polycythemia
-Newborns, High altitude,
Hypoxia due to lung and
heart diseases.
Congestive Heart failure,
Burns (loss of plasma),
Dehydration, Severe
Exercise, Emotional stress
DECREASED PCV
Anaemia
Pregnancy
(Hemodilution)

32. RED BLOOD CELL INDICES:RED BLOOD CELL INDICES:
1.Mean corpuscular volume(MCV)
2.Mean corpuscular hemoglobin(MCH)
3.Mean corpuscular hemoglobin concentration(MCHC)
4.Red cell distribution width (RDW)

33. Mean Corpuscular
Volume(MCV)
Average or mean volume of a single red blood cell
Expressed in femto liter (fl)

34. Calculation
Formula:
MCV = PCV in percentage X 10
RBC count per cmm
Normal range:
80-100 fl

35. INCREASED
Megaloblastic anaemia
Chronic alcoholism
Liver disease
newborns
DECREASED
Microcytic hypochromic
anaemia

36. Mean Corpuscular
Hemoglobin(MCH)
Average hemoglobin content (weight of Hb) in a
single red blood cell
Expressed in picograms(pg).

37. Calculation:
Formula:
MCH = Hb in gm/dl X 10
RBC count per cmm
Normal range: 27 – 32 pg

38. INCREASED
Macrocytic anaemia
Newborns
DECREASED
Microcytic anaemia

39. Mean Corpuscular Hemoglobin
Concentration(MCHC)
Concentration of haemoglobin in 1 dl or 1 liter of packed
red cells

40. Calculation
Formula:
MCHC = Hb in gm/dl X 100
PCV in %
Normal range: 30 – 35 g/dl

41. INCREASED
Hereditary spherocytosis
DECREASED
Hypochromic anaemia

42. CLINICAL SIGNIFICANCE
A) Macrocytic anaemia:
◦MCV slightly increased upto 150 fl
◦MCH is slightly increased
◦MCHC is normal or diminished
B) Microcytic anaemia:
◦MCV is diminished up to 50 fl or lower
◦MCH is diminished to 15 pg or lower
◦MCHC is diminished to 20% or less
C) Spherocytosis:
◦MCV is diminished
◦MCHC is elevated

43. RED CELL
DISTRIBUTION WIDTH
Measures the degree of variation of red cell size in
a blood sample.
Increased in iron deficiency anaemia
Decreased in beta- thalassemia trait
Normal value: 9.0-14.5