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Rapid polymer-based immunohistochemistry for intraoperative CNS tumor diagnosis: A validation study

Julia Schuler, Sandra Baur, Federico Fusco, Felix Schicktanz, Benedikt Wiestler, Julian Canisius, Christian Mawrin, Denise Bernhardt, Arthur Wagner, Bernhard Meyer, Carolin Mogler, and Claire Delbridge

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Volume 45 (2026) p. 98 - 106

Abstract

Clinical Neuropathology, Vol. 45 – No. 3/2026 (98-106)

Rapid polymer-based immunohistochemistry for intraoperative CNS tumor diagnosis: A validation study

Julia Schuler¹, Sandra Baur¹, Federico Fusco¹, Felix Schicktanz¹, Benedikt Wiestler², Julian Canisius², Christian Mawrin³, Denise Bernhardt⁴, Arthur Wagner⁵, Bernhard Meyer⁵, Carolin Mogler¹, and Claire Delbridge¹

¹Institute of Pathology, ²Department of Neuroradiology, TUM Klinikum, TUM School of Medicine and Health, Technical University of Munich, Munich, ³Department of Neuropathology, Neurosurgery, and Neurology, Otto-von-Guericke University, Magdeburg, ⁴Department of Radiation Oncology, and ⁵Department of Neurosurgery, TUM Klinikum, TUM School of Medicine and Health, Technical University of Munich, Munich, Germany
Intraoperative diagnostics in neuro-oncology face a critical gap between the morphological limitations of hematoxylin-eosin staining or toluidine blue staining and the time-intensive nature of conventional immunohistochemistry (IHC). This delay hinders real-time surgical decision-making, particularly in distinguishing gliomas, metastases, and lymphomas. To address this, we validated a novel, rapid IHC protocol utilizing directly conjugated polymerized horseradish peroxidase (pHRP) antibodies, designed to deliver results in under 30 minutes. We evaluated the performance of pan-cytokeratin (pan-CK), glial fibrillary acidic protein (GFAP), and cluster of differentiation 20 (CD20) antibodies on 115 intraoperative frozen tissue samples. Staining quality was assessed using a 4-tiered scoring system (from 0 to 3+). The cytoplasmic markers GFAP and pan-CK demonstrated excellent robustness and reliability, achieving mean scores of 2.6 and 2.47, respectively, with over 68% of cases showing strong, specific staining (3+). The membrane-bound marker CD20 showed more variability (mean score 2.07), highlighting the influence of antigen localization on this rapid protocol. Our findings demonstrate that this rapid IHC method is a viable and highly effective tool for the intraoperative differentiation of central nervous system tumors, particularly for cytoplasmic antigens. By providing crucial morphological information in near real-time, this approach has the potential to significantly enhance diagnostic precision during surgery, enabling immediate, tailored therapeutic strategies and improving patient outcomes.Correspondence to:
Dr. med. Claire Delbridge
or
Julia Schuler
Institute of Pathology
TUM School of Medicine and Health
Technical University of Munich
Munich, Germany
Email: [email protected]; [email protected]

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