Abstract

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.

Author Details

Authors

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

Departments

• ¹ Institute of Pathology,
• ² Department of Neuroradiology, TUM Klinikum, TUM School of Medicine and Health, Technical University of Munich, Munich,
• ³ Department of Neuropathology, Neurosurgery, and Neurology, Otto-von-Guericke University, Magdeburg,
• ⁴ Department of Radiation Oncology, and
• ⁵ Department of Neurosurgery, TUM Klinikum, TUM School of Medicine and Health, Technical University of Munich, Munich, Germany

Address

Dr. med. Claire Delbridge, Institute of Pathology, TUM School of Medicine and Health, Technical University of Munich, Munich, Germany, or Julia Schuler, Institute of Pathology, TUM School of Medicine and Health, Technical University of Munich, Munich, Germany
Email: [email protected]

Citation