Single Cell and Immunoanalysis Core

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Project Summary/Abstract: The Single Cell and Immunoanalysis Core Research into human skin biology and inflammatory skin diseases has been slowed by the lack of transgenic model systems and the limited samples that can be obtained from living patients. However, advanced analytic techniques that provide maximal information from small samples have been developed that have the potential to advance our understanding of human skin disease. This Core makes critical groundbreaking techniques including single cell RNA sequencing (scRNA-seq) with the ability to batch samples, discover TCR genes and tag cells with antibodies (CITE-seq), as well as single cell ATAC-seq, single cell imaging by flow cytometry, cytometry by time of flight (CyTOF) and high throughput T cell receptor sequencing available to investigators at any institution who wish to carry out human skin disease research. The Center also provides assistance with experimental planning, key reagents and assistance with data analysis for each technique. For example, the Center provide protocols that allow skin biopsies to be shipped from distant locations and viably cryopreserved before scRNA-seq studies; this advance makes single cell profiling available to investigators to study samples from clinical trials and from understudied skin diseases. The Center also provides CyTOF staining protocols, validated panels of CyTOF antibodies, custom conjugation of CyTOF antibodies, advice on experimental design and data analysis, and all services needed for outside investigators to send their cells to the Center for CyTOF studies. The Center provides comprehensive support in the experimental design, DNA extraction protocols, DNA sample preparation services, shipping, and data analysis assistance that will enable researchers to successfully utilize high throughput TCR sequencing to profile skin immune responses and track pathogenic T cell clones across multiple timepoints and tissues. We have included 36 research projects from investigators who wish to utilize the services of this Core. In summary, this core provides access to single cell approaches that have the potential to provide unprecedented insights into the biology of human skin disease.