Artifacts in Histopathology
Artifacts in Histopathology
Artifacts in Histopathology
179]
Review Article
Abstract Histopathological examination is considered as gold standard procedure for arriving at a final diagnosis of
various lesions of the human body. However, it is limited by a number of alterations of normal morphologic
and cytological features that occur as a result of presence of artifacts. These artifacts may occur during
surgical removal, fixation, tissue processing, embedding and microtomy and staining and mounting
procedures. They can even lead to complete uselessness of the tissue. It is therefore essential to identify
the commonly occurring artifacts during histopathological interpretations of tissue sections. This article
reviews the common artifacts encountered during slide examination alongside the remedial measures which
can be undertaken to differentiate between an artifact and tissue constituent.
Address for correspondence: Dr. Syed Ahmed Taqi, Department of Oral and Maxillofacial Sciences, Division of Oral Pathology, Najran University College of
Dentistry, Najran, Saudi Arabia.
E‑mail: [email protected]
Received: 22.04.2017, Accepted: 27.02.2018
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Figure 3: Histopathological image shows tissue tear due to rough Figure 4: Histopathological image shows tissue tear and folds due to
handling by forcep (H&E, ×10) rough handling by forcep (H&E, ×10)
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proteins and vacuolization of cytoplasm. Nuclear changes They are removed from tissue sections by immersion in
include pyknosis, karyolysis and karryohexis [Figure 7].[7] saturated alcoholic picric acid. The use of buffered neutral
formalin will minimize this problem.[9]
Improper prefixation
Solutions such as normal saline do not fix the tissue and Mercury pigments
tissue will undergo autolysis. Microscopically, such a section Mercuric chloride containing fixative usually, but not
will show features of autolysis artifact as well as separation invariably, produces dark brown granular deposits. Mercuric
of epithelium from the connective tissue (simulate chloride pigment is extracellular and can be removed by
vesiculobullous lesions such as pemphigus) [Figure 8].[8] treatment in alcoholic iodine.[9]
Figure 5: Histopathological image shows crush artifact (H&E, ×10) Figure 6: Histopathological image shows sutural artifact (H&E, ×10)
Figure 7: Histopathological image shows tissue autolysis due to Figure 8: Histopathological image shows separation of epithelium from
delayed fixation (H&E, ×10) the connective tissue simulating vesiculobullous lesion (H&E, ×10)
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Decalcification
Acid‑decalcifying fluids have hydrolytic actions that can
cause digestion of cellular and other tissue components
more rapidly when the tissue is unfixed or partially fixed.
Impairment or loss of tissue basophilia, inactivation of
enzymes, loss of iron and ribonucleic acid and destruction
of tooth enamel can occur. This can be reduced by Figure 9: Histopathological image shows bone trabeculae stained
thorough fixation of specimen before decalcification.[13] strongly with hematoxylin due to incomplete decalcification (H&E, ×10)
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Chatters
Chatters refer to thick and thin zones parallel to the knife
edge.
Figure 10: Histopathological image shows cholesterol clefts in Figure 11: Histopathological image shows scoring and tearing of
odontogenic cysts (H&E, ×10) section due to nick in knife edge (H&E, ×10)
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Air bubbles
As the tissue sections are flattened in the water bath,
bubbles of air may become trapped beneath them.
Collapsed bubble artifact occurs due to collapsing of
air bubbles entrapped beneath the sections leaving
cracked areas when dry, which fail to adhere to the
glass slide properly and show altered staining. This can
be prevented by using freshly boiled water in floatation
bath.[20]
Staining artifacts
Residual wax
Failure to remove the wax from parts of a section before Figure 16: Histopathological image shows wrinkles and folds due to
uneven stretching of tissue sections (H&E, ×10)
staining will result in incomplete or partial staining in that
area. This can result in the Pink disease artifact. This artifact
was studied in detail by Nedzel et al. in 1951. This author
described the presence of intranuclear birefringent (paraffin
wax) inclusions, particularly in the lymphocytes.[21]
b. Highly refractile lines outlining cells and tissue 3. Margarone JE, Natiella JR, Vaughan CD. Artifacts in oral biopsy
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This artifact can be corrected by removing coverslip in applications. Br Dent J 2004;196:329‑33.
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cytologic specimens. Oral Surg Oral Med Oral Pathol 1985;60:195‑6.
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processing on the content and integrity of nucleic acids. Am J Pathol
Artifact related to excessive use of mounting media 2002;161:1961‑71.
Excessive use of mounting media or the mounting media 8. Abbey LM, Sweeney WT. Fixation artifacts in oral biopsy specimens.
will result in foggy appearance. This artifact can be Va Dent J 1972;49:31‑4.
9. Pizzolato P. Formalin pigment (acid hematin) and related pigments. Am
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CONCLUSION 2008. p. 53‑105.
11. Okun MR, Ellerin P, Piotrowicz MA. Prevention of ice crystal damage
Tissue artifacts can be introduced into tissue specimen to biopsy specimens in transport. Arch Dermatol 1972;105:458‑9.
12. Carleton HM, Drury RA, Wallington EA. Carleton’s Text Book of
during any one of the many steps through which a specimen Histological Techniques. 5th ed. Vol. 41. The University of Michigan:
is carried before its microscope features are examined by Oxford University Press; 1967. p. 269.
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with prompt fixation and careful tissue processing will bone. J Histochem Cytochem 1962;10:560‑3.
14. Krishnanand PS, Kamath VV, Nagaraja A, Badni M. Artefacts in oral
minimize the artifacts. The first article regarding artifacts mucosal biopsies: A review. J Orofac Sci 2010;2:57-62.
was published by Zegarelli in 1978. Since then, very few 15. Chatterjee S. Artefacts in histopathology. J Oral Maxillofac Pathol
articles appeared on this subject.[23] Through this review 2014;18:S111‑6.
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Vol. 78. London, Boston: Butterworths; 1985. p. 18, 78, 611.
that prevent proper diagnosis as well as suggested some 17. Culing CF, Allison RT, Barr WT, Culling CA. In microtome and
techniques of minimizing these problems. microtomy. Cellular Pathology Techniques. 4th ed. United Kingdom:
Oxford University Press; 1985.
Financial support and sponsorship 18. Bindhu P, Krishnapillai R, Thomas P, Jayanthi P. Facts in artifacts. J Oral
Nil. Maxillofac Pathol 2013;17:397‑401.
19. Woods AE, Ellis RC, editors. Laboratory Histopathology: A Complete
Reference. Edinburg: Churchill Livingstone; 1994.
Conflicts of interest 20. Niemann TH, Tranovich JG, De Young BR. Biopsy bag artifact. Am J
There are no conflicts of interest. Clin Pathol 1998;110:224‑6.
21. Faoláin EO, Hunter MB, Byrne JM, Kelehan P, Lambkin HA, Byrne HJ,
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