THE BLOOD SMEAR

Material, steps, results

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- CBC – COMPLETE BLOOD COUNT

- gives important information about the kind and number of cells in the blood, especially
red blood cells, white blood cells, and platelets.
- helps health professional check any symptoms, such as weakness, fatigue, or bruising.
- helps diagnose conditions, such as anemia, infection, and many other disorders

- WBC – WHITE BLOOD CELL COUNT

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MATERIALS AND METHOD

- MATERIALS
- medium size blood drop – from finger tip or peripheral vein, on K3EDTA
- grinded glass slide for blood drop spreading
- support glass slide – on which the smear will be performed – clean, no grease
- staining kit – methanol/eosine/azure-methylene/distilled water
- slide support for drying

- METHOD
- performing and staining the blood smear
- microscopic examination
- normal values – absence of morphological variations
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METHOD
- preparation
- sterile gloves, sterile needles, puncture area disinfection
- harvesting
- medium size blood drop
- from finger tip or peripheral vein, on K3EDTA
- remove the first 1-2 drops by dry cotton – use the 3rd drop.
- spreading
- take the 3rd drop by a grinded slide edge
- spread the blood drop on a clean slide
- a standard smear should include
- size 3 cm length/ 2 cm width
- uniformly spread
- short terminal fringes < 0,3 cm
- reading area on the middle – 1-1,5 cm
- thick area at the smear onset

- fixing
- drying at room temperature
- completing the fixation by anhydrous methanol 10 sec
- staining
- eosin staining 20 sec – stain the cytoplasm and Hb
- azure methylene 30 sec – stain the proteins and nucleic acids
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ERITROTOPOIEZA METODA FROTIULUI DE SÂNGE

blast = young cell

cit = evolved cell
Round nucleus in early stages
meta = after
Stem cell RBC (mature erythrocyte)– no nucleus
pro = before

Proerythroblast

Basophilic Erythroblast

Polychromatic Erythroblast

Acidophylic Erythroblast
Expulsion Reticulocyte Erythrocite
FORME PATOLOGICE DE ERICTROCIT METODA FROTIULUI DE SÂNGE

Reversibile shapes Spherocytes

Echinocytes (burr cells)

• Ereditary spherocytosis
• Autoimune hemolitic anemia
• Severe burns
• Transfusions
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Reversibile shapes
Eliptocytes Stomatocytes

• pH<7
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Reversibile shapes
Knizocyte

• hemolitic anemia
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Ireversibile shapes

Acantocytes (spurr cells) Drepanocytes

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Citoplasma

Cytoplasm vs
Cell type Picture Size (μ) Color Granules Nucleus
nucleus

Neutrophil Many, small multilobulated shape

9-15 Pale-pink More cytoplasm Primary – light gray-blue 2-5 segments
(granulocyte) Secondary – red-brown Purple-red

Eosinophil Bi-lobulated
Many, big
12-17 Pink More cytoplasm
Red-orange intense
2 segments
(granulocyte) Dark-red

Basophil Little, big partially lobulated

10-14 Blue More cytoplasm
Dark blue, black Dark blue
(granulocyte)

Monocyte Very little Various shapes

15-30 Gray-blue Little more cytoplasm Dark-red (round, C, U, kidney)
(agranulocyte) - AGRANULOCYTE - Purple-red

7-12 Very little

Limphocyte (small); Sperical
Light-Blue More nucleus Dark-red
(agranulocyte) 12-16 Dark blue
- AGRANULOCYTE -
(large)
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Cell type Main function

Respiratory gases transportation (O2 from lungs to tissue and CO2 from tissue to
Erythrocyte lungs)

Neutrophyl Chemotaxis, phagocytosis, anti-microbial function

Role in parasitic infections response, predominant inflammatory cells in allergic
Eosinophyl reactions

Allergic and antigen response by releasing the chemical histamine causing

Basophyl inflammation

Chemotaxis, phagocytosis, foreign microoganism destruction by migrationg from

Monocyte blood to tissue and turning into macrophage

Thrombocyte Major role in blood clotting

Lymphocyte Immune response

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Cell type Normal values Lifespan Percent value

(from all leucocytes)

Erytrocites Male: 4,5-5,5 million/mm3 110-120 days

Female: 4,0-5,0 million/mm3
Newborn:4,0-7,0 million/mm3
Child: 4,0-6,0 million/mm3

Leucocytes Adult: 4 000 - 9 000 /mm3

Newborn: 15 000 - 25 000 /mm3

Neutrophyls 1,5-7,5*109/l t1/2 almost 7 h 50-75 %

Eosinophyls 0,02-0,60*109/l t1/2 almost 6 h 1-4 %

Basophyls 0,01-0,15*109/l 0-1 %

Monocytes 0,2-0,8*109/l t1/2 almost 70 h 4-8 %

Lymphocites 1,2-3,5*109/l 25-35 %

Thrombocytes 130 000 - 300 000 /mm3 9-12 days

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X 200
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X 1000
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X 1000
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