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Immunocytochemistry on the Thinprep processor
D J Dabbs1, C S Abendroth, R T Grenko
1Department of Pathology, Pennsylvania State University, College of Medicine, Hershey 17033, USA.
Insights
Immunocytochemistry (ICC) on ThinPrep Processor (TPP) slides showed equal or greater staining intensity and distribution compared to direct smears. TPP offers advantages like cleaner backgrounds and adaptability for multiple antibody tests.
Area of Science:
- Cytopathology
- Immunohistochemistry
- Diagnostic Pathology
Background:
- Immunocytochemical studies (ICC) are crucial for diagnosing non-gynecologic cytologic specimens.
- The ThinPrep processor (TPP) is approved for non-gynecologic specimen preparation, but its utility with ICC is understudied.
- Existing literature lacks comprehensive data on ICC methodology applied to TPP-processed cytology specimens.
Purpose of the Study:
- To evaluate the efficacy of immunocytochemistry (ICC) on specimens processed using the ThinPrep Processor (TPP).
- To compare ICC staining results from TPP-prepared slides with traditional direct smears (DS).
- To assess the advantages and limitations of TPP for ICC in cytopathology.
Main Methods:
- Collected 41 fresh surgical specimens for ThinPrep (TL) and direct smear (DS) preparation.
- Utilized both Papanicolaou-stained (P) and unstained (US) slides, with some US TL slides undergoing antigen retrieval (AR).
- Performed ICC using a panel of 33 antibodies and graded staining semiquantitatively, with each sample serving as its own control.
Main Results:
- ThinPrep slides (TL) demonstrated equal or superior staining intensity and distribution compared to direct smears (DS).
- TLUS slides showed better staining than DSUS slides in 44% of cases.
- No significant difference in immunostaining was observed between Papanicolaou-stained and unstained TL slides; antigen retrieval offered no advantage.
Conclusions:
- ICC on TPP-processed slides is effective, yielding results comparable or superior to direct smears.
- TPP offers advantages including cleaner backgrounds, reduced antibody usage, and flexibility for multiple antibody tests.
- TPP is a viable and advantageous method for performing ICC on cytologic specimens.
Introduction:
For cytologic specimens, the vast majority of immunocytochemical studies (ICC) are performed on non-gynecologic specimens for diagnostic purposes, and they can be performed on unstained or previously stained direct smears. Although the ThinPrep processor (TPP) has been approved for the preparation of non-gynecologic specimens, there is scant literature describing the utility of ICC methodology on cytology specimens fixed and processed by this method.
Materials And Methods:
Forty-one fresh specimens were obtained from the surgical gross room and aspirated or scraped to collect cells for thin layer (TL) and direct smears (DS). Specimens included a variety of neoplastic and nonneoplastic samples that were either Papanicolaou (P) stained or unstained (US). One group of US TL slides was subjected to antigen retrieval (AR). Staining was graded semiquantitatively. Each sample acted as its own control. Antibodies (abs) included: CAM5.2, AE1/3, K903, vimentin, MSA, desmin, s-100, HMB45, PSA, PAP, chromogranin, NSE, insulin, synaptophysin, pCEA, mCEA, mCEAD14, LCA, L26, UCHL-1, OPD-4, thyroglobulin, GCDFP, ER/PR, laminin, collagen IV, PLAP, HCG, CD68, HAM56, and MAC387.
Results:
Semiquantitative staining overall results comparisons: TLP > DSP TLP < DSP TLP = DSP TLUS > DSUS 11/25 (44%) 6/25 (24%) 8/25 (32%) 9/24 (38%) TLUS < DSUS TLUS = DSUS 3/24 (12%) 12/24 (50%) TLP Vs. TLUS TLP > TLUS TLP < TLUS TLP = TLUS 8/41 (20%) 9/41 (22%) 24/41 (58%) There were five false-negative results, 2 with TL and 3 with DS, and 1 false-positive TL.
Discussion:
Immunocytochemistry performed on the ThinPrep Processor showed equal or greater intensity and distribution of proper staining when compared to direct smears with the following advantages: (1) cleaner background, easier to interpret; (2) less abs required in a smaller area; (3) IPX can be done on Papanicolaou-stained thin layer slides; (4) thin layer slides can be modified for multiple abs tests; (5) additional thin layer slides can be prepared for ICC bases on needs. No significant differences of immunostaining were seen when comparing thin layer Papanicolaou-stained and unstained slides. Antigen retrieval offered no advantage in this study.