2. Blood film is examined for
• DLC
• General assessment of cell counts
• RBCs morphology
• WBCs morphology
• Platlet morphology
• Study of other defects like rouleaux
formation, agglutination, red cells
inclusions etc
3. Preparation
• Place a small drop of blood on central line
1 cm from one end
• Place a spreader at 45o
angle in front of
blood
• Push the spreader forward by the rapid
,smooth and straight movement
• Allow the film to dry in air
4. Staining
• Most commonly used stains are
• Giemsa
• Jenner’s Stain
• Wright’s stain
• Leishman’s stain
5. Giemsa Staining
• Fix air dried blood film with methanol for 5-10 minutes
• Transfer the film to a jar containing Giemsa stain(1:10
dilution) for 10-15 minutes
• Wash with buffered water
• Allow to stand in a jar containing buffered water for 3-5
minutes
• Dry it in a vertical position
Examination:
• First examined under low power(*10) objective to get an
idea of quality of film
• Then select a suitable area and examine with(*40)
objective
12. Abnormalities of Colour
• Hypochromia- IDA and Thalasemia
• Hyperchromia-
• Spherocytes
• Macrocytes
• Target Cells: Central hemoglobin area surrounded by a
pale ring and then hemoglobin in peripheral area
• Dimorphism :Two populaions of RBCs in the same
blood film-one normal and other abnormal
• Seen in –sideroblastic anemia
• After blood transfusion
• Patients receiving hematonics
21. Abnormalities of Shape
2.Elliptocytes and ovalocytes
• 10% seen in normal blood fil(tail end)
Seen in :
• Iron deficiency anemia(pencil Cells)
• Megaloblastic Anemia
• Myelofibrosis(tear drop cells)
23. Abnormalities of Shape
3.Stomatocytes:
• Mouth like slit in RBCs
• Few in normal blood film
• Increase in alcoholism
• Liver disease
• Hereditary membrane defects
24. Abnormalities of Shape
4.Schistocytes:
• Fragmented RBCs of various shapes and
sizes e.g helmet cells
Seen in:
• IDA
• Megaloblastic anemia
• Thalasemia
• DIC
• Microangiopathy
27. Abnormalities of Shape
5.Echinocytes and burr cells:
• RBCs with evenly distributes blunt
spicules. Burr cells are echinocytes with
reversible spicules
Seen in
• after prolonged standing of blood at room
temp
• when film is made on an oily slide
29. Abnormalities of Shape
6.Acantocytes
• RBCS with fewer thorn like projection of
varying sizes and variable in number
• more blunted than Echinocytes.
Seen in
• Disorders of liquid metabolism
• chronic liver disease ] Acquired.
• Spur cell anemia
33. Inclusions in RBCS
1.Hb crystals : Hb c and Hbs form crystals inside
the RBCS
2.Howell – Jolly bodies: small rounded , reddish to
blue fragments of nucleus ,<1 um in diameter .
May be single or multiple
• splenectomy
• splenic atrophy
• alcoholism
• sickle cell anemia and megaloblastic anemia
35. Inclusions in RBCS
3. Basophilic stippling or punctuate
basophilia :
• Fine to coarse , deep blue to purple , small
and multiple inclusions of varying sizes.
• Represent aggregated ribosomes.
• 1. Thalasemia 2. megaloblastic anemia
3.liver disease 4. lead poisoning
5.Infections.
37. Inclusions in RBCs
4.Pappenheimer bodies;
• small , dark staining , irregular granules of
hemosiderin , near the periphery.
• Seen in
• 1. Sideroblastic anemia 2.thalasmia and
3. dyserythropoietic anemia.
38. Inclusions in RBCs
5.Cabot Rings:
• Thin, reddish blue,ring like structure.it may
be twisted to form structure of 8
• Seen in: Severe anemia, megaloblastic
anemia, lead poisoning, dyserythropoeitic
anemia
• Parasites e.g malaraial parasite
