How to make a blood smear, including all steps: Preparation, fixation and staining. Veterinary clinical Examination for small and large animals. Methanol - Fixation. Additionaly basic information about blood.

1. 1. Basic Understanding of Blood
2. Blood-Smear-Test
• Preparation
• Fixation
• Staining
Clinical Diagnostic
Lukas K.
3rd academic year
October 2016

2. 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

3. 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

4. Stem cell can become any
red/white blood cell
Stem cell
Resting & Activated
PlateletLymphocyte
Monocyte
Neutrophil
Erythrocyte

6. 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

7. 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
Microscopy

8. 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.

9. Let´s have a look at the Blood Smear Test

10. Blood Smear Test
Preparation
Fixation
Staining
Includes 3 steps

12. 2. Fixation of Blood Smear

13.  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.

14. Different fixatives
Methyl alcohol
(methanol) is
the choice,.
although ethyl alcohol
("absolute alcohol") can
be used

15. Can you distinguish between a good done blood
smear from a bad one?
Let´s compare some

16. The shape of blood film

17. 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.

18. 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:
1. If the haematocrit increased, the angel of the spreader slide
should be decreased.
2. If the haematocrit decreased, the angel of the spreader slide
should be increased.

19. large angle
low HCT
small angle
high HCT

20. Parts of Blood Smear
A 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.

21. Common cause of a poor blood smear:
1. Drop of blood too large or too small
2. Spreader slide pushed across the slide in a jerky manner
3. Failure in keep the entire edge of the spreader slide against the slide while
making the smear
4. Failure in keep the spreader slide at a 30⁰ angel with the slide
5. Failure to push the spreader slide completely across the slide
6. Irregular spread with ridges and long tail: edges of spreader dirty or
chipped ; dusty slide
7. Holes in film – slide contaminated with fat or grease and air bubbles
8. Cellular degenerative changes: delay in fixing inadequate fixing time or
methanol contaminated with water

22. 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.

23. Examples of unacceptable smears

24. 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.

25. Even with naked eye a blood smear can
already indicate different physiological
conditions of organism