blood smear preparation, fixation, staining. veterinary clinical examination of small animals and...
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1. Basic Understanding of Blood
2. Blood-Smear-Test• Preparation
• Fixation• Staining
Clinical DiagnosticLukas K.3rd academic yearOctober 2016
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Blood is soup
Liquid portion
Vegetables- Water
- Salts (electrolytes)- Small proteins- Antibodies
- Red blood cells- White blood cells- Platelets
These cells are produced in the bone marrow from larger,
more complex progenitor cells.
Through a series of divisions, these larger cells are transformed into red blood cells,
white blood cells, and platelets
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Stem cell
Lymphoid Stem Cell
Myeloid Stem Cell
Resting & Activated Platelet
Lymphocyte
Monocyte
Neutrophil
Erythrocyte RBC
WBC
Series of divisions transforms complex cells into red blood cells,
white blood cells, and platelets
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Stem cell can become any red/white blood cell
Stem cell
Resting & Activated PlateletLymphocyte
Monocyte
NeutrophilErythrocyte
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Immunology
Immunology
Immunology
Immunology
Immunology
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What Is a Blood Smear? A blood smear is a blood test used to look for abnormalities in
blood cells. The three main blood cells that the test focuses on are:
•white cells, which help your body fight infections
The Vegetables
•red cells, which carry oxygen throughout the body
•platelets, which are important for blood clotting
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White Blood Cell
Each cell type has a specific function in the body. The forms of the cells
are specialized to handle these functions.
Red Blood Cell
Platelet
Electro Microscop
y
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What has the shape of the cells to do with the blood smear test?
Number of Blood cells Shape of Blood cells
Blood-Smear-Test is a very cheap and easy blood examination test and is done in sveral minutes, he basically provides us simple information of
which can ”help” doctors diagnose certain blood disorders or other medical conditions.
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Let´s have a look at the Blood Smear Test
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Blood Smear Test
PreparationFixation Staining
Includes 3 steps
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2. Fixation of Blood Smear
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To preserve the morphology of the cells, films must be fixed as soon as possible after they have dried.
Why do we fixate blood smears?
It is important to prevent contact with water before fixation is complete.
Methyl alcohol (methanol) is the choice, although ethyl alcohol ("absolute alcohol") can be used.
Methylated spirit (95% ethanol) must not be used as it contains water.
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Different fixativesMethyl alcohol
(methanol) is the
choice,.
although ethyl alcohol ("absolute alcohol") can be
used
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Can you distinguish between a good done blood smear from a bad one?
Let´s compare some
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The shape of blood film
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Qualities of a Good Blood Film
It should not cover the entire surface of slide.
It should have smooth and even appearance.
It should be free from waves and holes.
It should not have irregular tail.
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Want to change thickness of Blood Smear?
1. The angle of the spreader slide. (the greater the angle, the thicker and shorter the smear).
2. Size of the blood drop.3. Speed of spreading.
Notes:4. If the haematocrit increased, the angel of the spreader slide
should be decreased.5. If the haematocrit decreased, the angel of the spreader slide
should be increased.
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large angle
low HCT
small angle
high HCT
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Parts of Blood SmearA peripheral blood film consists of 3 parts :
1. Head: the portion of blood film near the drop of blood.2. Body: the main part of the blood film.3. Tail: the tapering end of the blood film.
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Common cause of a poor blood smear:
1. Drop of blood too large or too small2. Spreader slide pushed across the slide in a jerky manner3. Failure in keep the entire edge of the spreader slide against the slide
while making the smear4. Failure in keep the spreader slide at a 30 ⁰ angel with the slide5. Failure to push the spreader slide completely across the slide6. Irregular spread with ridges and long tail: edges of spreader dirty or
chipped ; dusty slide7. Holes in film – slide contaminated with fat or grease and air bubbles8. Cellular degenerative changes: delay in fixing inadequate fixing time
or methanol contaminated with water
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A. Blood film with jagged tail made from a spreader with achipped end.
B. Film which is too thick C. Film which is too long, too wide, uneven
thickness and made on a greasy slide.D. A well-made blood film.
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Examples of unacceptable smears
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Biologic causes of a poor smear
1. Cold agglutinin - RBCs will clump together.Warm the blood at 37° C for 5
minutes, and then remake the smear.
2. Lipemia - holes will appear in the smear. There is nothing you can do to
correct this.
3. Rouleaux - RBC’s will form into stacks resembling coins. There is nothing you can do to correct this.
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Even with naked eye a blood smear can already indicate different
physiological conditions of organism
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Hope you learned something