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ARTEFACTS

CONTENTS
Introduction
Classification
Types of artefacts
Remedies
Summary
References
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INTRODUCTION

Artefact is referred to as an artificial structure


or tissue alteration in a prepared microscopic
section which is a result of an extraneous
factor
Importance of biopsy specimen
Tissue from body - processed
Microscope by pathologists
Various chemicals processing and staining
Artefacts produced at various stages
Clinician induced defects
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Diagnosis

ARTEFACTS

2 principle circumstances
artefacts
I)Handling of the tissue at the time of biopsy or
during fixation process
II) Histo technical procedures used for embedding
and staining the tissue

ARTEFACTS

Classified/ categorized according to the stage


in preparation of the section, from the
removal of the tissue from the body to the
processing of the tissues for examination.
Some overlap b/n the groups exist and it is
not always possible to say which stage or
procedure was responsible for a given
appearance
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ARTEFACTS

Artefacts created during biopsy


Fixation artefacts
Tissue processing artefacts
Artefacts during microtomy
Staining artefacts
Cover slip/ mounting artefacts
Artefacts in cytology
Artefacts of resin bound tissues
Artefacts in immunohistochemistry
Artefacts during frozen sections

ARTEFACTS

Biopsy is the removal of part, or all, of a lesion to


enable histopathological examination and
definitive diagnosis
Biopsies establish evolutive control of disease
processes, and are able to document healing or
relapse

Clinician induced artefact


Heat artefact
Crush artefact
Monsels solution
Injection artefact
Pseudo-cyst formation
Specimen marking dyes
Foreign body artefacts

Heat artefact
Caused by laser generated heat
Heated metal instrument
Over heated molten wax,
Heating on hot plate

Crush artefact

Tissue distortion resulting


from tissue compression
caused
by improper handling of
tissue by forceps
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ARTEFACTS

Necrosis due to Monsels solution

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Intra-lesional injection
of anesthetic solution
causing vacuolization

Section showing pseudocyst formation


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Foreign body artefact


Silk sutures
Gel foam artefact
Tatoo pigments
Starch contamination
Cellulose contamination

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Silk suture artefact

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Tatoo artefact

Starch contamination

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Cellulose artefact

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Artefacts during Transport

Specimen containers
Rust
Specimen marking dyes
Foam sponges
Freezing artefacts

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Specimen
marking dye

Foam sponges/ biopsy pads

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Freezing artefacts

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Artefacts due to specimen contamination

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Artefacts during Fixation

Curling artefact
Formalin haem pigment
Formalin pigment
Improper fixation

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Curling artefact

Streaming
artefact

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Formalin-Haeme
pigment artefact

Improperly fixed
tissue

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Improperly fixed
tissue
Autolysis and
putrefaction

Fixation in water or
saline

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Fixation in mercuric chloride

Formalin pigment

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Artefacts during Processing

Various steps in processing


Dehydration
Clearing
Impregnation\ infiltration
Embedding

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Poor Processing

Loss of soluble
structures

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Artefacts during Microtomy

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Coarse chatter

Holes from roughing


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Sectioning artefact Knicks formed


in tissue due to faulty blade

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Venetian blind

Knife lines

Displacement of
components
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Bubbles under
section

Tidemark due to
adhesive
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Dirt

Folds

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Artefacts during Staining

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Artefacts during Staining

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Contaminated
staining solution

Residual
wax

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Mucosal
contamination

Water
contamination
Drying of the
section
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Artefacts during Mounting\ Cover


slipping

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Fungal
contamination

Cellulose fibers

Presence of
hair
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Sneezing artefact
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Air bubble seen in section due to improper


mounting technique or improper mountant solution

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Mountant
breakdown artefact

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Bleaching artefact

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Foreign body artefact caused


due to presence of a strand of cotton wool

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Artefacts in routine histopathology


Clinician induced artefact How do they appear

Causes and Remedies

Use of preparation sites

If possible are to be
avoided

Interfere with tissue


processing and
staining procedures

Improper surgical
removal

Pemphigus vulgaris
longer, broader,
shallower biopsy
specimen is required
Verrucous carcinoma
take a narrow deep
specimen
epithelium should be
thick to see for the
dysplastic changes if
deep seated.

Split artefact
Fragmentation
artefact

Scalpel blades
splitting of both
superficial and at the
edge of the tissue
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During traction by the

Artefacts in routine histopathology


Clinician induced artefact How do they appear

Causes and Remedies

Pseudocyst formation

Numerous spaces
lined with surface
epithelium false
impression of cysts in
connective tissue

Teeth of the
instrument force
epithelium through
the connective tissue

Heat artefacts/
fulgration

Amorphous
appearance to the
epithelium and the
connective tissue
Epithelial cells appear
detached, fusiform,
nuclei show
hyperchromatism,
spindled, palisading
configuration
Connective tissue and
muscle appear
opaque and
amorphous

Coagulation of the
tissue protein
Avoided by making
the incision margin far
away from the
interface of the lesion
with the normal tissue
Prevent accidental
contact of the cutting
tip with the metal
instruments to hold
the specimen
Initial incision with
scalpel

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Artefacts in routine histopathology


Clinician induced artefact

How do they appear

Causes and Remedies

Injection artefact

Haemorrhage with
extravasation mask
the normal cellular
architecture
Seperation of
connective tissue
bands with
vacuolization

Injection of anesthetic
solution directly into
the lesion
Injection deep to the
lesion or immediately
after performing biopsy
is effective for
haemostasis

Forceps/ crush/
squeeze artefact

Loss of cytological and


nuclar dimensions and
relations are affected

Grasping with
excessive force teeth of
the instrument
penetrate the tissue
Adequate retraction
and visualization
minimal force in
grasping
Use of small Adsons
forceps with or without
teeth l
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Lesion proper should

Artefacts in routine histopathology


Clinician induced artefact

How do they appear

Causes and Remedies

Starch artefact

Starch granules
resemble epithelial
cells
Refractile, glassy,
polygonal, PAS +ve
cells or spore like
structures
A central dark area
pyknotic nucleus or a
mitotic figure
Light blue in H & E
Blue black in Lugols
solution
Deep iliac red with PAS
Resistant to diastase

Starch powder from the


surgical gloves

Specimen containers

Crush/ tear/
compression

Small mouthed jars/


bottles
Wide mouthed flat
bottomed containers

Freezing artefact

Interstitial vacuoles

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Temp 12.2F or -11C

Artefacts in routine histopathology


processing artefact

How do they appear

Causes and Remedies

Fixation

Shrinkage
Curling
Brittleness
Pigmentation

Placing in fixative and


retaining its invivo shape or
placing the specimen on a
cardboard to maintain its
shape before immersion in
formalin
Deeper elliptical biopsies
avoided

Foam sponges

Triangular artefacts
Misinterpreted as
vascular spaces

Wrapped in perm paper

Processing
a) Dehydration

Over dehydration
brittle
Interfere with staining
properties
Under dehydration
hard to section
tearing artefacts and
holes in the sections

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Artefacts in routine histopathology


Artefacts during processing How do they appear

Causes and Remedies

Embedding

Uneven staining and


spreading
Wrinkle formation
Moth eaten effect
Venetian blind

Inadequate infiltration
insufficient time in
molten wax
Vibration of the tissue
in the wax block coz of
air entrapment
Vacuum oven used
Eliminated by soaking
in the block in moist
cloth

Microtomy

Cracks, streaks,
compresion artefacts
Squamous cell
contamination highly
eosinophilic and have
large nucleus
resembling skin
epidermal cells
Scratches appear as
straight slashes or
ragged tears

Dull or blunt knife


Rubbing of the
technicians fingers
Flaws or dirt on the
cutting edge

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Artefacts in routine histopathology


Artefact

How do they appear

Causes and Remedies

Incomplete staining
Precipitation
Contaminated taining
solution
Mucous contamination

Absence of the stain


Deposits
Microorganisms

Inadequately filled
staining dish
Filtering or freshly
prepared stains
Using slide staining jars
Washing of the slide

Preservation artefact
Water in section
Mountant break down

Minute bubbles
entrapped in the nuclei
Leaching of the stains
Crazing or cracking
crystallization
Bleached area

Remounting the section


Washed in a clearant
Polysterene based
mountants
Remount
Exposure to light for
excessive periods
Stored in darker areas

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Artefact Decalcifications

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Incomplete
decalcification artefact

Bone dust artefact

Over decalcification
artefact
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Limitations of paraffin
wax embedding for dense
bone artefact

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Artefact cytology

Propellent contamination

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Cardboard imprint

Drying

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THANK YOU

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