Artifacts in histopathology: An experimental study
Introduction
Histopathology is the art and science dealing with identification and diagnosis of lesions on stained representative tissue sections under the microscope produced after a series of sensitive laboratory procedures. It is of prime importance to identify unwanted artifacts and correlate them to the faulty laboratory step and avoid erroneous slide interpretation.
Aims
To correlate artifacts in hematoxylin and eosin stained tissue sections to various errors in fixation, processing and staining.
Methodology
It was a double blind study where, tongue specimens were collected from sacrificed pigs and various fixation, processing and staining errors were performed. Two observers reported the slides individually and the findings were correlated to the errors performed in the laboratory.
Results
Macroscopically tissue was hard and gritty when fixed using higher concentration of formaldehyde (50 and 100%) and 100% isopropyl alcohol (IPA). The tissue specimens were soft and friable with separation of epithelium from the connective tissue evident when immersed in saline and water. It was impossible to section tissues when processed in weak concentration of IPA (25 and 50%) and when clearing step was skipped.
Conclusion
Identification of artifacts is an important aspect in the practice of histopathology to avoid misinterpretation in reporting. As the available literature in this subject is sparse, this experimental study is an attempt to understand the effects of faulty laboratory procedures on the final stained tissue sections. These artifacts can also act as indicators of faulty laboratory techniques and guide us towards ideal standardized operative procedures in laboratory.
Histopathology is the art and science dealing with identification and diagnosis of lesions on stained representative tissue sections under the microscope produced after a series of sensitive laboratory procedures. It is of prime importance to identify unwanted artifacts and correlate them to the faulty laboratory step and avoid erroneous slide interpretation.
Aims
To correlate artifacts in hematoxylin and eosin stained tissue sections to various errors in fixation, processing and staining.
Methodology
It was a double blind study where, tongue specimens were collected from sacrificed pigs and various fixation, processing and staining errors were performed. Two observers reported the slides individually and the findings were correlated to the errors performed in the laboratory.
Results
Macroscopically tissue was hard and gritty when fixed using higher concentration of formaldehyde (50 and 100%) and 100% isopropyl alcohol (IPA). The tissue specimens were soft and friable with separation of epithelium from the connective tissue evident when immersed in saline and water. It was impossible to section tissues when processed in weak concentration of IPA (25 and 50%) and when clearing step was skipped.
Conclusion
Identification of artifacts is an important aspect in the practice of histopathology to avoid misinterpretation in reporting. As the available literature in this subject is sparse, this experimental study is an attempt to understand the effects of faulty laboratory procedures on the final stained tissue sections. These artifacts can also act as indicators of faulty laboratory techniques and guide us towards ideal standardized operative procedures in laboratory.
