An Improved Approach to Prepare Human Brains for Research

We describe two protocols for preparing human brains collected for research and diagnosis. In both protocols, one half brain is processed for research and the other for neuropathological evaluation. Clinical, neuropathological and tissue mRNA retention data are used for sample categorization. In protocol</it> /, coronal, whole hemisphere slices cut at standardized landmarks are frozen with a cooling device at −90°C, which yields discrete anatomical structures. In selected instances, small blocks of brain are frozen at −160°C in liquid nitrogen vapor. Cooling device or liquid nitrogen vapor frozen samples are suitable for in situ</it> hybridization, protein blotting or immunohistochemistry. Morphological freezing artifacts are minimal. In protocol 2</it>, one half brain is frozen en bloc</it> on dry ice; this tissue is suitable for regional evaluation of gene expression or neurochemistry. Morphological freezing artifacts are severe. In both protocols, the other half brain is fixed in formalin prior to sectioning and diagnostic evaluation. The standardized selection of paraffin blocks from each brain allows precise diagnoses to be established, including identification of dangerous infectious processes; moreover, it makes it possible to produce a set of uniformly selected blocks and slides for comparative studies. These protocols lead to standardized tissue preparation for research and reduce variables impairing interpretation and comparison of data.