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Immunocytochemical staining of unstained versus previously stained cytologic preparations
1Department of Pathology, College of Medicine, Pennsylvania State University, Hershey, USA.
Insights
Immunoperoxidase staining on various fine needle aspiration smear preparations, including previously Papanicolaou-stained slides, yields comparable diagnostic results. These methods are equally useful for adjunctive immunocytochemical studies in cytodiagnosis.
Area of Science:
- Cytopathology
- Immunohistochemistry
- Surgical Pathology
Background:
- Immunoperoxidase (IP) staining aids cytodiagnosis using fine needle aspiration (FNA) specimens.
- IP staining can be performed on unstained smears, cell blocks, or previously stained Papanicolaou smears.
- The adequacy of IP staining on previously stained smears requires further investigation.
Purpose of the Study:
- To compare the efficacy of IP staining on different FNA smear preparations.
- To evaluate IP staining on unstained, alcohol-fixed/air-dried, Papanicolaou-stained, and decolorized Papanicolaou-stained smears.
Main Methods:
- Compared IP staining results using antibodies against keratin, epithelial membrane antigen, CEA, desmin, actin, vimentin, leukocyte common antigen, B/T cell markers, and PSA.
- Utilized 75 sets of smears from surgical specimens, grading results semiquantitatively.
- Evaluated expected positive staining, background staining, and cellularity preservation.
Main Results:
- All tested methods demonstrated comparable results for expected positive staining.
- Background staining and cellularity preservation were similar across all preparation techniques.
- No significant differences were observed in the utility of the different smear preparations.
Conclusions:
- IP staining on unstained, alcohol-fixed/air-dried, Papanicolaou-stained, or decolorized Papanicolaou-stained FNA smears are of equal utility.
- These methods provide comparable results for adjunctive immunocytochemical studies in cytodiagnosis.
Abstract:
Immunoperoxidase (IP) staining of fine needle aspiration specimens may provide useful adjunct information in cytodiagnosis. IP may be used on unstained direct smears or cell block material if available. In the absence of such material, IP is sometimes used on previously stained (Papanicolaou) smears. Few studies have examined the adequacy of such preparations, however. We compared IP staining results on unstained, alcohol-fixed or air-dried smears versus alcohol-fixed, previously Papanicolaou stained smears and versus alcohol-fixed, decolorized, Papanicolaou-stained smears, using antibodies to keratin (CAM 5.2, AE1/AE3 and K903), epithelial membrane antigen, monoclonal carcinoembryonic antigen, desmin, muscle-specific actin (HHF-35), vimentin, leukocyte common antigen, B and T cell markers L26 and UCHL-1, and prostate-specific antigen. Seventy-five sets of smears were made from various neoplastic or nonneoplastic tissues received as surgical specimens, and antibodies were selected for anticipated positive staining. The results were graded in a semiquantitative fashion. All the methods gave comparable results regarding expected positive staining, background staining and preservation of cellularity. We conclude that these methods of smear preparation are of equal utility in adjunctive immunocytochemical studies.