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Modified rapid immunohistochemical staining for intraoperative diagnosis of malignant brain tumors
Akane Suzuki1,2, Takashi Maruyama1,3, Masayuki Nitta3
1Faculty of Advanced Techno-Surgery, Tokyo Women's Medical University, 8-1 Kawada-Cho, Shinjuku-Ku, Tokyo, 162-8666, Japan.
Insights
A modified rapid immunohistochemistry (R-IHC) technique enhances specimen stability and diagnostic accuracy for brain tumors. This improved method offers reliable results comparable to permanent specimens, aiding in faster and more precise tumor diagnosis.
Area of Science:
- Neuropathology
- Molecular Pathology
- Diagnostic Pathology
Background:
- Rapid immunohistochemistry (R-IHC) is crucial for timely central nervous system tumor diagnosis.
- Existing R-IHC methods face challenges with specimen preparation and immunostaining instability.
- A need exists for a more robust and reliable R-IHC technique.
Observation:
- A modified R-IHC protocol was developed using 4% paraformaldehyde fixative and a Polymer Refine Detection Kit.
- The modified R-IHC was evaluated on 94 brain tumor cases, assessing markers like GFAP, p53, pHH3, ATRX, CD20, and CD3.
- Stability and stainability were compared against standard R-IHC and permanent specimens.
Findings:
- The modified R-IHC demonstrated superior specimen stability compared to standard R-IHC for GFAP and p53 markers.
- Stainability achieved by the modified method was comparable to permanent specimens.
- High percentages of correct classification (PCC) were observed: 86.6% for pHH3, 82.8% for ATRX, and 100% for CD20/CD3 discrimination in lymphomas and gliomas.
Implications:
- The modified R-IHC technique offers enhanced reliability and accuracy for rapid brain tumor diagnosis.
- This advancement can improve diagnostic workflows, leading to quicker treatment decisions.
- The method shows potential for broader application in neuropathological diagnostics requiring rapid immunostaining.
Abstract:
Rapid immunohistochemistry (R-IHC) has been developing mainly as a support technique in the rapid diagnosis of central nervous system tumors; however, there have been problems regarding instability in specimen preparation and immunostaining. To overcome the weakness of this technology, the instability of immunostaining, we developed a modified R-IHC. This was achieved by switching to 4% paraformaldehyde as the fixative solution and utilizing a commercially available Polymer Refine Detection Kit, as a high-sensitivity kit, in place of the secondary antibodies. In this study, we tested the modified R-IHC by evaluating rapid immunostaining on new staining items in 94 brain tumor removal cases, which took place at Tokyo Women's Medical University from 2014 to 2015. The results showed that, based on GFAP and p53 markers, the modified method obtained a higher stability in specimens than the standard rapid immunostaining method. It also achieved stainability on the same level as that of a permanent specimen. The modified method tested 86.6% (46/53) and 82.8% (24/29) in pHH3 and ATRX, respectively, in the percentage of correct classification (PCC) against the permanent specimens, and 100% (7/7) in the PCC against malignant lymphomas and gliomas that used CD20/CD3 for discrimination. We concluded that the modified R-IHC method indicated a higher stainability and PCC against the permanent specimens in comparison to the standard method in GFAP, p53, CD20/CD3, pHH3, and ATRX.
