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  • Protocol for the Systematic Fixation, Circuit-Based Sampling, and Qualitative and Quantitative Neuropathological Analysis of Human Brain Tissue.

Protocol for the Systematic Fixation, Circuit-Based Sampling, and Qualitative and Quantitative Neuropathological Analysis of Human Brain Tissue.

Methods in molecular biology (Clifton, N.J.) (2022-11-19)
Caitlin S Latimer, Erica J Melief, Jeanelle Ariza-Torres, Kim Howard, Amanda R Keen, Lisa M Keene, Aimee M Schantz, Trevor M Sytsma, Angela M Wilson, Thomas J Grabowski, Martin Darvas, Kristen Dams O'Connor, Amber L Nolan, Brian L Edlow, Christine L Mac Donald, C Dirk Keene
摘要

Human brain tissue has long been a critical resource for neuroanatomy and neuropathology, but with the advent of advanced imaging and molecular sequencing techniques, it has become possible to use human brain tissue to study, in great detail, the structural, molecular, and even functional underpinnings of human brain disease. In the century following the first description of Alzheimer's disease (AD), numerous technological advances applied to human tissue have enabled novel diagnostic approaches using diverse physical and molecular biomarkers, and many drug therapies have been tested in clinical trials (Schachter and Davis, Dialogues Clin Neurosci 2:91-100, 2000). The methods for brain procurement and tissue stabilization have remained somewhat consistently focused on formalin fixation and freezing. Although these methods have enabled research protocols of multiple modalities, new, more advanced technologies demand improved methodologies for the procurement, characterization, stabilization, and preparation of both normal and diseased human brain tissues. Here, we describe our current protocols for the procurement and characterization of fixed brain tissue, to enable systematic and precisely targeted diagnoses, and describe the novel, quantitative molecular, and neuroanatomical studies that broadly expand the use of formalin-fixed, paraffin-embedded (FFPE) tissue that will further our understanding of the mechanisms underlying human neuropathologies.

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Sigma-Aldrich
抗-亨廷顿蛋白抗体,克隆mEM48, culture supernatant, clone mEM48, Chemicon®
Sigma-Aldrich
抗-Tau(3-重复亚型 RD3)抗体,克隆8E6/C11, culture supernatant, clone 8E6/C11, Upstate®
Sigma-Aldrich
抗-Tau(4-重复亚型 RD4)抗体,克隆1E1/A6, culture supernatant, clone 1E1/A6, Upstate®
Sigma-Aldrich
抗朊病毒蛋白抗体,a.a.109-112,克隆3F4, clone 3F4, Chemicon®, from mouse
Sigma-Aldrich
抗-NOTCH 3/N3ECD抗体,克隆1E4, clone 1E4, 1 mg/mL, from mouse