Examine peripheral blood film

1. Prepared by
HadeelAl Sadoun
Examine peripheral blood film

2. How you examine Blood film
 Macroscopic Examination
 10x objective Examination
 40x objective Examination
 100x objective – oil Immersion Examination

3. Macroscopic Examination
 Examine the smear before placing on the microscope.
(Why?)
 To indicate abnormalities:
 Example: (over all blurred smear  increased blood proteins
:multiple Myeloma)

4. Microscopic Examination
 Microscope should be properly adjusted on the area of blood
smear will be examined.
 The light from the illuminator should be properly centered.
 Condenser should be placed all the way up.
 Iris diaphragm must be opened

5. 10x objective Examination
 This magnification power must not be skipped for the following
reasons:
 Quick scanning of over all smear quality
 Assess the distribution of the RBCs.
 AssessWBC distribution
 Large abnormal cells can be seen example:
 Blast
 Reactive lymphocytes
 Unexpected parasites

6. 40x objective examination
 Easy to select the examination area as well as 10x objective
examination.
 WBC estimation
 The examiner should select an area where two or more RBC
overlapped and all other are evenly separated
 Count theWBC number regardless of there type in 10 fields
 Use this equation:
 Average no. ofWBC/ field x 2000= cells/ µL
 NormalWBC estimate ranges from : 5,000 – 10,000 cell/µL

7. 100 objective- oil immersion
Examination
 Used forWBC differential count
 Classify 100WBC into (PMN, Lymphocytes, Monocytes, eosinophiles and
basophiles)
 For Platelets estimate and morphology
 Count platelet in 10 fields under 100x.
 Use the following equation:
 Average no. of Platelets/ field x 20,000 = cell/ µL
 Normal range of platelets = 150,000 – 350, 000 cell/ µL
 RBC morphology and nucleated RBC examination and count

8. Basophiles
Esionphils
Monocytes
Lymphocytes
PMN
10-14µm
12-17µm
12-18µm
varies
12-14µm
Bilobed with large
dark blue granules
may obscure the
nucleus
Bilobed with large
red-orange
granules.
Nucleus folded, or
kidney shaped,
chromatin is less
dens than
lymphocytes.Very
fine azurophilic
granules.
Large:12-16µm,
nucleus is round ,
coarse chromatin
Small: 9-12µm,
nucleus is round ,
coarse
chromatin, High
N:C ratio.
Multilobed (2-4)
nucleus connected
by thin strands
(0-2%)
(1-4%)
(1-10%)
(18-42%)
(48-70%)

9. Platelet morphology
 Normal platelets:
 Size: 1-3 µm
 Only seen under 100x objective magnification.
 Irregular outlines
 fine- violet granulation (scattered or centralized)
 Giant platelets:
 Size: 4-5 µm
 Same features of morphology

10. RBC Morphology
 Normal RBC:
 Size: 6.8- 7.2 µm
 central pallor, not vaculated or graulated.
 Abnormalities in size:
 Microcytic RBC: size less than 6µm
 Macrocytic RBC: size more than 8 µm
 Variation in RBC sizes called anisocytosis

11.  Abnormalities in hemoglobin concentration:
 Hypochromic: pale RBC less hemoglobin available than normal
 Hyperchromic: dark red RBC , concentrated Hemoglobin

12. RBC morphology
 Abnormal shapes of RBC:
 Stomatocyte
 Target cell
 Eleptocyte
 Variation of the cell shape in field called poikilocytosis

13. How to evaluate RBC, WBC and platelet
morphology
 If abnormal cell present:
 Examine 10 fields:
 If 0-2 cell seen in 10 field : +
 If 2-4 cells seen in 10 field: ++
 If 5-7 cells seen in 10 field : +++