Optimizing immunohistochemical neuropathological protocols for human brain banking

Accurate neuropathological diagnosis is essential for human brain banking, yet inconsistent immunohistochemical (IHC) protocols across different banks have long compromised diagnostic reliability and sample comparability.

Using formalin‑fixed, paraffin‑embedded archival tissues from the China Brain Bank of Zhejiang University, Juan‑Li Wu and colleagues systematically optimized four critical stainings for hyperphosphorylated Tau, β‑amyloid, α‑synuclein, and phosphorylated TDP‑43. Through comparative testing of antigen retrieval conditions, they identified optimal parameters: for p‑Tau and p‑TDP43, boiling in citric acid buffer (pH 6.0) for 15 minutes; for α‑synuclein, boiling in Tris‑EDTA buffer (pH 9.0) for 15 minutes followed by 80% formic acid for 10 minutes; and for β‑amyloid, boiling in citric acid buffer (pH 6.0) for 15 minutes plus 80% formic acid for 10 minutes, with the 6F3D antibody clone delivering the clearest signals.

Importantly, after optimization, staining results from the China Brain Bank of Zhejiang University were fully consistent with those from the Netherlands Brain Bank, a global leader in open‑access brain banking. This cross‑validation confirms the reliability, repeatability, and international compatibility of the optimized protocols. The study fills a critical gap in the China Human Brain Bank Consortium’s standard operating procedures, providing a much‑needed unified IHC framework for quality control and accurate neuropathological diagnosis. This work, entitled “Optimizing Immunohistochemical Neuropathological Protocols for Human Brain Banking” was published on Human Brain (published on Apr. 1, 2026).