Human brain tissue collection and preservation for cortical neuronal cell type classification

The research project described in this plan is critically important to achieving an Allen Institute goal to characterize the functional architecture of the human neocortex at the level of cell types and local circuits and create a publicly available scientific resource of this information to advance the understanding of the brain. Such characterization includes molecular, anatomical and physiological analysis at the single cell level, using tissues from human brain.
 

More specifically, characterization is expected to include:

• Transcriptional analysis (e.g., PCR, RNA sequencing).
• Filling of single neurons with dyes (e.g., biocytin, Lucifer yellow) for morphological reconstruction and quantitative anatomy.
• In vitro slice electrophysiology for physiological characterization and correlated analysis of functional, morphological and molecular properties of individual cells, and for studies of population dynamics using calcium-sensitive dyes.
• Analysis of the molecular composition of synapses (e.g., array tomography).
• Extended maintenance of cortical tissues in culture for virally-mediated molecular genetic studies of circuit function.

In this collaboration, the expertise and resources of University of Washington will be applied in the areas of consenting, scheduling, arrangement of tissue collection, provision of facilities for minimal tissue processing, and management of IRB approvals, to provide human brain tissue and de-identified patient information to the Allen Institute.

Investigators from both parties will work together to establish appropriate exclusionary criteria, determine scientifically useful patient information that can be shared with the Allen Institute, establish requirements for minimal Institute use of UW Neurosurgery facilities for tissue processing, and establish logistical and administrative processes that enable rapid communication, transfer and processing of tissue to meet tissue viability needs for robust characterization of neuronal cell types.

The parties anticipate that the collaboration will result in profiling of tissues on a regular basis throughout the year, up to once a week.