Add tags 3 Jan 2021, 8:46 AM

‫ﺑﺴﻢ ﷲ اﻟﺮﺣﻤﻦ اﻟﺮﺣﻴﻢ‬

University of Alfashir
Faculty of medical Laboratory science
Department of Haematology
&
immune-haematology

Quality control in preparation of

blood smear and staning

Prepared by/ Ayman Abdallah Ahmed

Supervised by / Mrs. Maysa Abdulhadi
1/Introduction to blood smear

A blood smear, also referred to as a peripheral smear

for morphology, is an important test for evaluating
blood-related problems, such as those in red blood
cells, white blood cells, or platelets. It has a wide
range of uses, including distinguishing viral infections
from bacterial infections, evaluating anemia, looking
for causes of jaundice, and even diagnosing malaria

2/Preparation of blood smear

STEP ONE:
Place clean glass slide on a flat surface. Add one small
drop of blood to one end.

STEP TWO:
Take another clean slide, and holding at an angle of
about 45 deg, touch the blood with
one end of the slide so the blood runs along the edge
of the slide by capillary action. Push
carefully along the length of the first slide to produce
a thin smear of blood.

STEP THREE:
Make 2 smears, allow to air dry, and label clearly.
Once dried place in the provided slide
transport containers.

3/Quality control of smearing

Making great blood films takes practice; don’t be

discouraged by less than perfect smears. Submit
all blood smears, perfect or otherwise, as areas
of the film may be suitable for examination.
• Collect blood in an EDTA tube and make the
smears when back at the clinic.

• Use clean, high-quality microscope slides.

• Aim for a blood droplet size of 4mm diameter.
• Optimise spreading speed for length and a
good feathered edge.
• Hold the spreader slide at 30-40 degrees to
achieve optimal smear length.
• Maintain even contact throughout the
spreading motion.

4/troubleshooting
For short smear
Use a larger droplet of blood.
Decrease the angle of the spreader slide.
Decrease the speed of the spreader slide.
For wavy and radegs smear
Maintain even contact and a smooth motion.
Increase the speed of the spreader slide.
Relax the wrist, reduce downward pressure on
the spreader slide.

5/staining of blood smear

The stained peripheral blood film is one of the world’s
most widely and frequently used tests. Since its
introduction in the late nineteenth century, basic
elements of the blood film preparation and analysis
have changed little. Modern technology improvements
and refinements have enhanced the availability of
good quality commercial Romanowski stains,
automated stainers and semi–automated slide
makers.

6/Staining Quality Control

Optimal results are obtained by fixing and staining

directly after the blood film is completely air-dried.
Fixation of blood films before staining is recom-
mended, although many laboratories have a practice
of staining immediately after air-drying of blood films.
This usually yields acceptable results. If slides
cannot be stained immediately, however, fixation in
methanol is necessary within4 hours, but preferably
less than or equal to 1 hour, after air-drying;
otherwise,the plasma causes gray-blue background
effects. Staining and fixing solutionsmust be free
from water as much as possible (-3%) to prevent
morphological artifacts. If manual staining procedures
are used it is generally recommended that slides be
immersed in reagent-filled (Coplin) jars rather than by
coveringslides with staining solution because this may
lead to the formation of precipitateby evaporation. If
staining is performed under extremely hot conditions,
caremust be taken to prevent evaporation during
staining, for instance by per-
forming staining in a closed jar or in a closed petri
dish.Automated staining devices have their own
specific staining procedures asprovided by the device
manufacturers. Users may modify these procedures
to satisfy local requirements in terms of cell staining.
Staining protocols varybetween laboratories and no
generally accepted routine staining method or
result is available. Guidelines for staining and staining
results can be found inhematology and laboratory
guidelines and textbooks. The International Council
for Standardization in Haematology has published a
reference staining methodfor blood films, based on
purified azure B and eosin Y solutions.21
As a general rule for judging the quality of a stained
blood film thelaboratory must ensure that all cell
types in a blood film can be identifiedreliably by the
staining procedure followed. This applies equally to
manual andautomated methods. Blood films are
typically stained by Romanowsky dyes
(consisting of a variety of thiazines and eosins). A
number of methods havebeen described and include
Wright, Wright-Giemsa, May-Gru¨nwald-Giemsa,
and Leishman.