Smear Report
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ScribdTranslationsUNIVERSITY IGUAÇU - UNIG
FACULTY OF BIOLOGICAL AND HEALTH SCIENCES - FaCBS
Pharmaceutical Sciences Course
Report of the practical class of biochemical analysis
Nova Iguaçu
2014
ALINE CARDOSO DE ARAUJO LIMA
TAMARA DO NASCIMENTO SANTOS
Report of the practical class of Homeopathy
Report required as part of
second evaluation in the subject
Biochemical Analysis, taught
by teacher XXXXXXX, in
course of Sciences
Pharmaceuticals of the University
Iguaçu.
New Iguaçu
2014
Summary
This report refers to the internship carried out at the Laboratory
discipline of the Iguaçu University, within the scope of biochemical analyses.
The internship had as its main objective the integration into the Service of
biochemical analysis of the University, providing the acquisition of
practical skills in executing laboratory techniques, always
associated with a theoretical knowledge that allows for the interpretation of
results.
The areas explored in this report are Immunology and
Hematology, with only a generic reference to Biochemistry and to
Microbiology.
1. INTRODUCTION
The practical class conducted had as its main objective the integration of
theory and practical analysis within the disciplinary laboratory of the University, under
the guidance of Professor XXXXX. This integration in the laboratory practice was
fundamental for the perception of all the responsibilities that one has
biochemical analyst. Thus, it was useful for the practical application of
scientific knowledge acquired throughout the classes.
The area of Biochemical Analyses is constantly expanding and
development is one of the fundamental areas within the sciences
of health. The techniques used are increasingly tending towards effectiveness,
thus ensuring the quality of the results.
This practice was carried out during the period of 1 class. The time was
distributed between the aspects of a theoretical and practical class.
The biochemical analysis procedures are referred to, such as
blood smear.
The preparation of the blood smear is undoubtedly the point
crucial for the achievement of a reliable blood count and therefore, its
standardization should be one of the main requirements of a good laboratory
hematology. The smears can be performed by hand (method of
preference, if it is necessary to make a smear in the field), the support
on the table or something like that can still be used, and are used
a blade (clean, without traces of grease or other materials) and a
transparent glass distensor (an extender can be assembled with a
small slide glued to a slide with adhesive tape). The ideal smear should be
free from failures and stops, not too thick, nor too thin, and without flaws in the
tail. In the microscopic observation, the two edges where the
Counts should show the erythrocytes more separated and the leukocytes well.
distributed.
2. OBJECTIVE
The objective of this experiment was to observe through the optical microscope.
the components of human blood and their characteristics.
3. MATERIAL
Lance or injection needle
Cotton
Rectified alcohol
Blades washed with coconut soap or detergent, rinsed in
distilled water, dried and placed in a glass with alcohol, sealed.
Test tube
Test tube rack
Optical microscope
Pipette
Sample
Obtained by puncturing the fingertip with a previously used lancet.
disinfected. The blood collection will be made considering that the
The drop of blood extracted must be placed on the inverted slide, which only touches
in the emerging drop, thus avoiding contact with the skin.
4. METHODS
Execution of peripheral blood smears
The peripheral blood smears were prepared and stained manually.
Procedure
EXECUTION OF THE SCRAPING
1. Mix the blood by gently inverting the tube. Remove the cap.
2. Withdraw a small amount of blood from the tube with the pipette and place a small
drop of blood on a blade.
3. With one hand, hold the blade at the opposite corners to the end that
contains the blood.
With the other hand hold another blade or lamella, which will be used to spread the
blood, and place over the first in such a way as to make an angle of about
30º. This angle is maintained during the execution of the smear.
4. Pull the upper blade/lamella back until it contacts the drop of blood.
The blood will spread along the edge of this blade.
5. Perform the smear by sliding the upper blade/lamella gently but
quickly along about 4 cm of the lower blade.
6. Air dry the smear.
Figure 1 - Blood smear technique
Source - PUC - RS
Conditions for a good smear
1. Thin, that is, the different globules must be spread out in a
single layer without overlap, nor grain or flake formation.
2. Complete: the drop of blood must be spread out in its entirety, hence that
it should not be big. It should be taken into account that in a swipe
the richest parts in cytological elements are the edges.
FIXATION
The smear was covered or the slide was introduced into a container with alcohol.
absolute ethanol P.A. for 3 to 5 minutes. Dried in still air.
EXAM
A 100x objective was used with immersion oil. In the smear of
thin layer, examined 'ZIG-ZAG' passing through the fringe.
The number of red blood cells ranges from 4.5 to 6 million per mm3 of blood, the
white blood cell count of 8,000 to 10,000 per mm3 of blood and that of platelets
from 150,000 to 400,000 per mm3 of blood.
COLORATION
Used the GIEMSA solution, diluted in the proportion of 2 drops of
Giemsa solution for each ml of buffer solution (or distilled water).
Introduced the slide into a container with the diluted GIEMSA solution, left
run for 30 minutes. The blade was washed in running water to remove the
excess dye. Dried in still air.
Action of the dyes
Eosin - Acid dye. Stains basic cellular components
designated by eosinophils or acidophils, orange.
Methylene blue - Basic dye. Stains acidic cellular components.
designated by basophils, of bluish purple.
Methylene blue - Cora granulations designated as granulations
azureophiles, in red.
Polychromatophilic cells - Grayish color due to their proportions
identical acidic and basic components.
Neutrophil granulations - Result from the combined action of eosin and
pinkish methylene blue.
Observation of peripheral blood smears
In the microscopic observation of the stained smear, are sought
aspects that justify or complement the information provided by
hemogram. Qualitative or quantitative alterations of the
white blood cells, red blood cells, or platelets, being referred to in the report
of analysis the indications that are deemed pertinent from a clinical point of view.
EXAMPLES of information obtained from the observation of the smear of
peripheral blood that may be indicated in the analysis report:
Study of platelets.
Presence of aggregation;
Anisocytosis and changes in platelet granulation;
Study of the leukocyte series.
Research on atypical cells;
maturation deviations;
morphological changes;
presence of blasts or other young forms.
Study of the erythrocytic series:
Anisocytosis - different sizes.
Anisocytosis - normochromic cells coexisting with hypochromic.
Polychromatophilia - cells more bluish and larger than a mature erythrocyte.
The bluish color is due to the fact that they have less Hb than a red blood cell.
mature and still have RNA remnants in the cytoplasm.
Poikilocytosis - different shapes, with no predominance of any particular one.
Predominant abnormal forms: dianyocytes; dacryocytes; spherocytes;
elliptocytes; acanthocytes; echinocytes; schistocytes; etc.
Erythrocytic inclusions:
Basophilic stippling - small dark blue structures similar to
points, evenly distributed throughout the hemoglobinized area
of erythrocyte. The stippling results from the precipitation of ribosomes and RNA.
during the coloring.
Cabot rings - structures in a ring or figure-eight shape that are colored by
reddish-purple. They result from remnants of nuclear membrane or from
a mitotic spindle.
Howell-Jolly bodies - small round inclusions of color
reddish, considerably larger than the basophilic stippling.
Usually found one per erythrocyte and deviated to one side
from the cell. They correspond to nuclear remnants (e.g. nuclear membrane).
5. RESULT/ DISCUSSION
In the experiment with the drop of blood on the slide, it was observed through
of optical microscopy, countless red blood cells, practically the same way and
some types of leukocytes, basophil granulocytes were observed and
neutrophil granulocytes (multiple nuclei). It was also observed that
presence of lymphocytes (large nucleus). Finally, one can observe a
platelet aggregate, whose details could hardly be observed.
6. CONCLUSION
With the practical class based on the microscopic observation of cells from
human blood tissue, it was possible to conclude that a little is needed
technique and practice so that the blood smear is satisfactory, and that the
dyes are of fundamental importance for visualization of the
leukocytes.
Observing the slide, one can notice that the red blood cells are
present in greater numbers than the other cells. The leukocytes are
the constituent cells of blood tissue that show greater
dimensions, although they are relatively few in number. These can
to be classified into different types, according to shape and dimensions
of the nucleus.
It was also concluded that the cells of the blood tissue are immersed.
no plasma, and the platelets in the stained smear tend to appear in
groups.
7. BIBLIOGRAPHY
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LIMA, A. Oliveira; SOARES, J. Benjamin; GRECO, J.P; GALEZZI, João
CANÇADO, J. Romeu. Laboratory Methods Applied to Clinic. 5th ed.
Guanabara Koogan, Rio de Janeiro.
MOURA, Roberto A. de Almeida. Collection of material for examinations
laboratories. Atheneu. São Paulo, 1998;
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