Intraoperative immunocytochemistry of cytologic scrape specimens. A rapid immunoperoxidase method for triage and
D J Dabbs1, L Hafer, C S Abendroth
1Department of Pathology, Pennsylvania State University College of Medicine, Hershey.
Insights
A rapid immunoperoxidase (IPX) method for cytologic scrapings offers a faster alternative to frozen sections for intraoperative diagnosis. This technique efficiently triages tumors for further specialized studies, saving valuable tissue samples.
Area of Science:
- Surgical Pathology
- Cytopathology
- Immunohistochemistry
Background:
- Intraoperative frozen sections are crucial for timely surgical diagnosis.
- Current methods can be time-consuming and consume small tissue samples.
- There is a need for rapid, tissue-sparing diagnostic techniques.
Purpose of the Study:
- To evaluate a rapid immunoperoxidase (IPX) staining method for cytologic scrapings.
- To assess its utility as an adjunct to intraoperative frozen sections.
- To determine its efficiency in triaging tumors for special studies.
Main Methods:
- Cytologic scrapings from fresh tissues and tumors were prepared on glass slides.
- Specimens were fixed in 95% alcohol and stained using the streptavidin-biotin technique.
- A panel of 14 antibodies, including keratins, vimentin, desmin, and LCA, were investigated.
- Staining was visualized using aminoethylcarbazole localization.
Main Results:
- The rapid cytologic IPX method yielded results in 15-20 minutes, comparable to frozen sections.
- Keratins, vimentin, desmin, and Leukocyte Common Antigen (LCA) showed optimal reactivity.
- The method is more rapid and convenient than frozen sections, preserving tissue for ancillary studies.
- HMB45 showed weak reactivity in one melanoma case.
Conclusions:
- Rapid cytologic IPX is a valuable adjunct for intraoperative diagnosis.
- It offers a faster and more tissue-sparing alternative to frozen sections.
- This method aids in the efficient triage of poorly differentiated tumors for special studies.
Abstract:
We investigated a rapid immunoperoxidase (IPX) method utilizing a wide variety of antibodies for use on cytologic scrapings of fresh tissue that were submitted for intraoperative frozen section. The handling, fixation and IPX staining of these cytologic scrape specimens are more rapid and convenient than frozen tissue sections and spare tissues that may be of small quantity for appropriate special diagnostic studies (flow cytometry, cell markers, genetic studies, electron microscopy, and so forth). Fresh tissues from normal organs and tumors were scraped with a scalpel blade, smeared on an uncoated glass slide and immersed in 95% alcohol for one minute. Antibodies investigated by this method included: CAM 5.2, AE1/3, keratin 903, desmin, vimentin, HHF-35, CEA, PLAP, NSE, S-100, HMB45, LCA, L26 and UCHL-1. Twenty-three fresh epithelial and mesenchymal tissues from the uterus, colon, tonsil, lymph node, kidney and adrenal gland were examined, along with 29 tumors. There were 21 carcinomas, 3 melanomas, 3 sarcomas and 2 lymphomas. Using the streptavidin-biotin technique with aminoethylcarbazole localization, IPX results were available in 15-20 minutes, comparable to the time required for the reporting of a frozen section diagnosis. Keratins, vimentin, desmin and LCA gave the best results. HMB45 was weakly reactive in one case. This rapid cytologic IPX method may be used as an adjunct to frozen sections and cytologic imprints for intraoperative diagnosis or for efficient triage of poorly differentiated tumors for special studies at the time of frozen section.
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