2. I.Erythrocyte sedimentation rate(ESR)
Rate of settling of RBC from the plasma after the addition
of anticoagulant.
Importance of ESR
1. Good index for the presence of hidden carcinoma but
active diseases.
2. It measures the suspension stability of RBC.
3. It measures the abnormal concentration of fibrinogen
and serum globulin.
Roleaux formation
(Packing or piling of RBC)
3. Methods
A.Wintrobe and landsberg method
Anticoagulant used- Ammonium potassium oxalate
(wintrobe solution/ double oxalate/balanced oxalate/
paul-Heller’s soln.)
Tube – wintrobe tube
- Left side – red; 0 on top and 10 cm bottom.
- Right side – white; 10 cm top and 0 bottom.
4. Procedure
1. With a long stem pasteur pipet, fill the wintrobe tube
with oxalated blood up to 0 mark.
2. Let the wintrobe tube stand perfectly vertical.
3. Read result after 1 hour. Reading must be done on the
left red side of the tube.
Normal values
Male - (0-9) mm/hr
Female - ( 0-20 )mm/hr ( bcg less RBC )
Children- (1-13) mm/hr
5. Red White
0 10
Layers
-Plasma layer
-Buffy coat (WBC and platelets)
-Packed RBC (hematocrit)
Wintrobe
tube
Red cells
at bottom 0 at bottom
10
6. Females have more space in settle down and faster than
male and children because they have less RBC.
( 1cm – 10mm/hr)
B.Westergren method (200mm) – most sensitive and
most accurate .
- Anticoagulant used -3.8% sodium citrate
- Tube- westergren tube (through suction method long
tube)
8. Procedure
1. Fill the tube with the citrated blood
2. Stand the tube vertically and read result at the end of the
1st hrs. and 2nd hrs.
Normal values
Male – (3-5) mm/hr
7-15 mm/2hr
Female – (4-7) mm/hr
12-17 mm/2hr
9. Comparision
Wintrobe Wester gren
Bore 3 mm 2.5 mm
Graduation up to 100 mm up to 200 mm
Anticoagulant Double oxalate 3.8% sodium
citrate
Amount of 1 ml 2.4 ml
blood
Reading once Twice
Hematocrit
10. C. Graphic cutler
Anticoagulant – 3.8% sodium citrate
D. Linzenmeier
Anticoagulant – 3.8% sodium citrate
E. Roarke- Ernstiene
Anitocagulant – Heparin
F. Bray’s
Anticoagulant- 3.8% sodium citrate
11. G. Micro methods
1) Micro landau
Anticoagulant- 5% sodium citrate
2) Smith micro
Anticoagulant- 5% sodium citrate
3) Crista or hellige- vollmer
Stages of ESR
1. Initial rouleaux formation – (first 10 min)
2. Period of rapid settling – (next 40 min)
3. Period of final settling – (last 10 min)
total 60 minutes or 1hr.
12. Factors in ESR
1. Intrinsic Factor
- nos of RBC ( less RBC faster settlement)
- size of RBC ( Bigger the size is faster the settlement)
- viscosity of Plasma ( less viscous fast settlement)
* nos of RBC- inversely
* size of RBC- directly
13. 2. Extrinsic factor
Length of tube ( smaller length fast settlement)
Diameter of tube (wider diameter fast settlement)
Position of tube(vertical or slightly fast settlement
Temperature ( high temp. fast settlement)
Pipetting ( incorrect pipetting result error)
Volume of blood ( less blood faster settle.)
Anticoagulant (more anticoagulant slow settlement)
14. II Osmotic fragility test
Test the stability of RBC in hypotonic solutions.
Follows the law of osmosis.
Factor affecting OFT
- Red cell shape
- Red cell volume
- Red cell surface Area
- State of Red cell membrane
*Fragile cells( decrease)- spherocytes
*Resistant cells( increase)- sickle cell , target cell,
reticulocytes
15. Methods
1. Sanford method
Different conc. Of hypotonic solution
12 test-tube is used
Initial solution used – 0.5% Nacl
Interpretation
No hemolysis – tubes with compact sediment and clear
solution.
Initial hemolysis -1st tube from the left with not so
compact sediment and with dark red solution
Complete hemolysis - 1st tube from the left without
sediment and with dark red solution.
19. 3. Griffin and Sanford method
4. Dacies method
Hemolysis read is used through spectrophotometer.
(Transparent – fake pink- light pink- red)
20. III.Erythrocyte Indices
Important in assessing borderline types of anemia.
Computed using 3 determinants Hb, hematocrit and RBC
count.
A. Mean corpuscular volume (MCV)
Average volume of an individual RBC.
volume % Hct x 10 = cubic micra or femtoliter
RBC in millions
Normal value- 82- 92 cubic micra.
22. B. Mean Corpuscular Hemoglobin (MCH)
Ratio of Hb to red cell count
Average weight or amount of Hb in an individual RBC in
millions
gm Hb x 10 = uug or picogram
RBC in million
Normal value – (27-33) uug
Interpretation
> 33 uug- macrocyte
< 27uug – microcyte
< 22 uug – microcytic hypochromic
Example
Hb = 16gm/100ml
RBC count = 5,500,000/cumm
MCH= 16 x 10 = 29 uug
5.5
23. C. Mean corpuscular hemoglobin conc.(MCHC)
Mean conc. Of Hb in the average RBC.
Normal Value = 32-38%
Average weight or amount of Hb in an individual RBC
gm Hb x 100 = %
vol. % Hct
Example
Hb -16 gm/ 100ml
Hct = 46 vol. %
MCHC = 16 x 1000 = 34.7%
46