A

SEMINAR PRESENTATION
ON
ARTIFACTS IN
CYTOPATHOLOGY
BY
ADAGA SAMUEL EKEH
12/04144004

OUTLINE

Introduction
Overview of cytopathology
Artifacts in cytopathology
Role of cytopathology
Overcoming artifacts in cytopathology
Conclusion.

INTRODUCTION

Artifacts are undesired objects or

phenomena influencing the appearance
of a smear and obstructing or even
preventing proper microscopic analysis of
diagnostic factors of a cytological sample.
They are usually products or formations
found in preparations of fixed tissues
caused by manipulations or reagents and
is not indicative of the actual structural
relationship.

INTRODUCTION CONTD

Artifacts in cytologic smears have been

considered as a potential diagnostic pitfall in
cytodiagnosis.
Several artifacts have been found to be
inherent to the cytological smears.
The accuracy of the cytologic examination
from any body site depends greatly on the
quality of collection, preparation, staining and
interpretation of the cytological material.
Inadequacy in any of these steps will
adversely affect the quality of diagnostic
cytology as a result of the presence of
misleading artifacts.

The three pre-requisites for a meaningful

cytodiagnosis are;
Proper technique: collection procedure,
preparation of smears, fixation and staining.
Microscopic examination of stained
smearspicture (history, clinical features,
physical examination, radiological and
laboratory findings).
Diagnostic cytology can be carried out by
different methods which includes the following;

METHODS OF CYTOPATHOLOGY
Diagnostic cytology can be carried out by different
methods which includes the following;

Exfoliative cytology: In this method, cells are

collected after they have been spontaneously
shed by the body or manually scrapped, brushed,
swabbed, aspirated or washed from the epithelial
surface of various body organs. In malignant
condition or during infection, the exfoliation
becomes exaggerated and the epithelial cells
show variation in morphology. Such exfoliated
cells when collected and appropriately stained,
give information on the epithelium from which
they are derived. These characteristic cellular and
nuclear appearances in cells thrown off from
healthy epithelium differ distinctly from those
derived from inflamed or malignant lesions.

METHODS CONTD

Fine needle aspiration cytology: This

is now a widely accepted diagnostic
procedure which has largely replaced
open biopsy. It is a technique used to
obtain material from organs that do not
shed cells spontaneously. Thus, it is
valuable in the diagnosis of palpable
lesions of the breast, thyroid gland, lymph
nodes and skin.
Sediment cytology: In this method, the
sample is collected from the fixative that
was used for processing the biopsy or
autopsy specimen

METHODS CONTD

The fixative is mixed properly in a centrifuge tube

and centrifuged. The sediment is used for
smearing. These sediments are cells that are
shed by the autopsy and biopsy specimen during
the processsing
Imprint cytology: This is indicated in the case of
tumours especially of lymph nodes. Soon after an
excision biopsy of lymph node, the specimen is
cut using a sharp scalpel blade. If there is blood
oozing from the outer surface, touch the surface
with a cotton ball soaked in normal saline. Then
take imprint smears by touching the cut surface
with a clean microscope slide and fix immediately

THE ROLE OF DIAGNOSTIC CYTOLOGY

Diagnostic cytology has been widely applied

as a diagnostic tool in the following areas;
As a screening tool to detect precancerous and
cancerous lesions.
It is also commonly used to investigate thyroid
lesions and diseases involving sterile body
cavities and a wide range of other body sites.
It is used for hormonal assessment in cases of
infertility in females.
It is used for genetic sex determination.
It aids in the diagnosis of certain infectious
diseases and other imflammatory conditions
(e.g florid tuberculosis, rheumatoid pleurisy).

ARTIFACTS IN CYTOLOGICAL
SMEARS

The presence of artifacts in cytological

smears is important, as this finding
although sometimes accidental may have
implications for diagnosis, prognosis and
therapy. However, on numerous
occasions the presence of these artifacts
which are varied in nature may give rise
to confusion and/or incorrect
interpretation

Artifacts found in routine cytological smears

can result from technical error in any of the
following stages of cytodiagnosis;
Methods of specimen collection.
Fixation and fixatives.
Preservation of fluid specimens prior to
processing.
Preparation of material for microscopic
examination.
Staining and mounting of the cell sample

The following artifacts can be found in routine cytological smears;

Fixation
artifacts
These artifacts
occur in too thick
cytological smears
resulting in
inadequate fixation
and stabilization of
the cellular
structures

Drying artifacts
These groups of artifacts are observed in
cytological smears stained with
haematological stains like May-Grunwald. In
this method, the smears are intentionally
air-dried, but if the smears are not correctly
These artifacts can be found in pap smears
requiring alcohol fixation when the time
between smearing and wet fixation is
prolonged resulting in rapid evaporation
and air-drying. Thus, from the instant the
smear is made, air-drying proceed
extremely rapidly, hence the urgency for
fixation. The cytoplasm takes up more eosin
and nuclear details are less clear resulting
in pale-stained nuclei, lack of differential
cytoplasmic staining, cytoplasmic and
nuclear eosinophilia

Crush/smearing artifacts
These artifacts are found in
cytological smears prepared
from specimens diluted
with blood. The specimen is
spread like a peripheral
smear and the particles
tend to come to the edge of
the smear. Crushing these
larger particles with undue
pressure can result in
crush/smearing artifacts

Iatrogenic artifacts

These artifacts are a group of

physician-related changes in
morphologic pattern involving
specimen acquisition
(instrumentation) or
therapeutic interventions such
as radiation, chemotherapy,
tissue ablation and surgery.
Such induced changes may
mimic those of neoplasia and
cause considerable diagnostic
confusion. Starch crystals,
suture materials and
lubricants are examples of
iatrogenic artifacts.

Pollen Grains
These are aerial
contaminants. They
generally present
with a thick refractile
cell wall.

Cornflake artifacts
This brown artifact is
thought to occur
when there is a delay
between removing
the slides from the
last xylene and
applying the
mounting media. This
may be due to air
trapped within the
surface grooves of
mature squamous
cells

Stain precipitate artifacts

These are highly amorphous and granular

precipitates seen in stained smears. They
result from improper staining procedures.
They are common if saturated solution of
stain is allowed to evaporate or dry up on
the smear (Ochei and Kolhatkar, 2000).

Worm-like artifacts

The presence of worms in cytological

smears is occasionally reported, although
various other structures exist that may be
confused with such parasites. Recognition
of these structures is important to avoid
overvaluation or confusion with true
worms. Morphological criteria are required
to differentiate these worm-like artifacts
regularly abserved in smears from clinically
significant parasites. These artifacts are
considered sample contaminants (intrinsic
contamination) or contaminants of
cytological smears during processing

OVERCOMING THE POTENTIAL

DIAGNOSTIC PITFALLS OF ARTIFACTS
In order to ensure accurate and reliable
cytodiagnosis from the microscopic
examination of an artifact-free cytological
smear, the following procedures are necessary.
1. Proper patient preparation prior to
sample collection:
Proper patient preparation is the beginning of
good cervical cytology. For instance, prior to
cervical smear collection, the patient should
be instructed before coming for smear
collection that:

She should not douche the vagina for at

least a day before the examination.
The patient should abstain from coitus for
one day before the examination.
Smear should not be taken during
menstrual bleeding, because of
contamination with blood, endometrial
component, debris and histiocytes. No
intravaginal drugs or preparations should
be used for at least one week before the
examination.

Strict adherence to standard operating

procedures in sample collection,
preservation, processing and
microscopic examination:
Immediate fixation of smears is essential.
Smears should never be allowed to dry
before placing the coverslip.
Haematoxylin should be filtered every day
before use.
All solutions and other stains are filtered
daily after use, to keep them free of
sediment.

Avoid contamination from one smear to

another.
Keep stains and solutions covered when
not in use.
All dishes are washed daily.
Stains are discarded and replaced as the
quality of the stain deteriorates.
Avoid contamination during placing of the
coverslip, with the dropper used to
dispense the mounting medium.
Dipping should be done gently to avoid
cell loss and the slide carrier should not
hit the bottom of the staining dish.

Agitation of the slides by occasional

dipping is necessary to remove excess
dye.
Agitation of the slides by occasional
dipping is necessary to remove excess
dye.
Dipping should be done gently to avoid
cell loss and the slide carrier should not
hit the bottom of the staining dish

3. Clinical correlation
Correlation of cytomorphology with the
clinical picture (history, clinical features,
physical examination, radiological and
laboratory findings) is important in order
to avoid misinterpretation of artifacts.

CONCLUSION
Diagnostic cytology has become a widely accepted
diagnostic tool in the diagnosis of malignant,
infectious, inflammatory, genetic and
endocrinological disorders. Artifacts resulting from
both intrinsic and extrinsic contamination of
cytological smears remain a major diagnostic
pitfall in cytodiagnosis. This has resulted in
increased difficulty in accurate microscopic
interpretation of cytomorphological structures in
cytological smears. familiarity with these artifacts
and strict adherence to standard operating
procedures is essential to avoid misinterpretations
that can lead to false positive or false negative
diagnosis.