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URL: https://dx.doi.org/DOI:10.17504/protocols.io.zxbf7in

Authors: Ryoji Amamoto, Emanuela Zuccaro, Paola Arlotta, Constance L. Cepko

Summary: Thousands of frozen, archived tissues from postmortem human central nervous system (CNS) are currently available in brain banks. As single cell and single nucleus technologies are beginning to elucidate the cellular diversity present within the human CNS, it is becoming clear that transcriptional analysis of the human CNS requires cell type specificity. Single cell and single nucleus RNA profiling provide one avenue to decipher this heterogeneity. An alternative, complementary approach is to profile isolated, pre-defined cell types and use methods that can be applied to many archived human tissue samples. Here, we developed FIN-Seq (Frozen Immunolabeled Nuclei Sequencing), a method that accomplishes these goals. FIN-Seq uses immunohistochemical isolation of nuclei of specific cell types from frozen human tissue, followed by RNA-Sequencing. We applied this method to frozen postmortem samples of human cerebral cortex and retina and were able to identify transcripts, including low abundance transcripts, in specific cell types.

Affiliations: Harvard Medical School, University of Padova, Harvard University, Harvard Medical School

External URL: https://www.biorxiv.org/content/10.1101/602847v1

Version: 1

Publication Date: 2019

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