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. 2021 Jun 15;14:161. doi: 10.1186/s12920-021-01011-z

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — A method for cryopreserving and processing adult human heart tissue for single nuclei RNA-sequencing. a Workflow for cryopreserving adult human heart tissue and subsequent isolation and processing of nuclei for use in 10× Genomics microfluidics platform. b Demographics of the four unused donor hearts.cBatch-corrected Uniform Manifold Approximation and Projection (UMAP) of the integrated dataset. Each dot represents a single nucleus and is colored by cluster identity. Clusters are numbered in descending order of the number of nuclei assigned to that cluster. d Separated UMAP of each individual donor dataset demonstrating that every cell type was present in each donor and batch effects have been removed. e Heatmap of the top ten differentially expressed genes in each cluster across all the clusters. Each row is a different gene, colored by the expression level of that gene in each individual nucleus, with each column representing an individual nucleus. The expression of all of the top ten genes for each cluster across all clusters is displayed for each nucleus in the dataset. The nuclei are grouped by cluster, and the colored bar at the top indicates cluster number and corresponds to the colors in c