@article{d7fdd626735c46d2bb6e16130edaeea4,
title = "Nuclei isolation from surgically resected human hippocampus",
abstract = "Single-nucleus RNA sequencing (snRNA-seq), where nuclear transcriptomes are a proxy to cellular transcriptomes, has been used to profile human brain. snRNA-seq is sensitive to tissue processing, tissue quality, postmortem interval time, and cellular debris. This protocol outlines steps for the isolation of high-quality nuclei from surgically resected human brain tissue followed by a sucrose gradient yielding neuronal and non-neuronal nuclei enabling unbiased analysis of various cell types. For complete details on the use and execution of this protocol, please refer to Ayhan et al. (2021).",
keywords = "Cell isolation, Cell separation/fractionation, Genomics, Microbiology, Neuroscience, RNAseq, Single Cell",
author = "Fatma Ayhan and Connor Douglas and Lega, {Bradley C.} and Genevieve Konopka",
note = "Funding Information: The authors thank the patients for consenting to participate in this research and Haley Moore for providing the images shown in Figure 1 A. G.K. is a Jon Heighten Scholar in Autism Research at UT Southwestern. This work was supported by the NIH ( NS106447 ), a UT BRAIN Initiative Seed Grant ( 366582 ), the Chilton Foundation , and the National Center for Advancing Translational Sciences of the NIH under the Center for Translational Medicine{\textquoteright}s award number UL1TR001105 to G.K. and B.C.L., the NIH ( T32DA007290 and T32HL139438 ) to F.A., the NIH ( NS107357 ) to B.C.L., and the Chan Zuckerberg Initiative, an advised fund of Silicon Valley Community Foundation ( HCA-A-1704-01747 ) and the James S. McDonnell Foundation 21 st Century Science Initiative in Understanding Human Cognition – Scholar Award ( 220020467 ) to G.K. Some images in the Graphical Abstract are reprinted with permission from Ayhan et al. (2021) . Funding Information: The authors thank the patients for consenting to participate in this research and Haley Moore for providing the images shown in Figure 1A. G.K. is a Jon Heighten Scholar in Autism Research at UT Southwestern. This work was supported by the NIH (NS106447), a UT BRAIN Initiative Seed Grant (366582), the Chilton Foundation, and the National Center for Advancing Translational Sciences of the NIH under the Center for Translational Medicine's award number UL1TR001105 to G.K. and B.C.L. the NIH (T32DA007290 and T32HL139438) to F.A. the NIH (NS107357) to B.C.L. and the Chan Zuckerberg Initiative, an advised fund of Silicon Valley Community Foundation (HCA-A-1704-01747) and the James S. McDonnell Foundation 21st Century Science Initiative in Understanding Human Cognition ? Scholar Award (220020467) to G.K. Some images in the Graphical Abstract are reprinted with permission from Ayhan et al. (2021). B.C.L. resected the hippocampal tissue. F.A. designed and optimized the protocol. C.D. assisted with tissue collection and banking. F.A. and G.K. wrote the manuscript. G.K. supervised the study. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2021 The Author(s)",
year = "2021",
month = dec,
day = "17",
doi = "10.1016/j.xpro.2021.100844",
language = "English (US)",
volume = "2",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "4",
}