Cytometry Core

点击翻译按钮获取中文摘要

Flow cytometry is an important tool to quantify and phenotype cells that define pathways relevant to diabetes. Previously limited to the analysis of circulating immune cells, advances in cell extraction from tissues has allowed flow cytometry to expand its capabilities to the study of developmental, metabolic and signaling programs in a wide variety of cell types. The Cytometry & Cell Sorting Core (CCSC) is a vital component of the Columbia University Diabetes Research Center (DRC), as it addresses the research needs of many DRC investigators who study immunological, developmental, and metabolic aspects of diabetes. The Core was created in 2010 in partnership with the Columbia Center for Translational Immunology and the Department of Medicine, with substantial University investment as well as NIH support (S10 and P30 grants) and expanded in 2017 by incorporating the Cancer Center’s flow cytometry core. The core expansion from 5 to 11 instruments (4 coming from the Cancer Center and 2 new spectral flow cytometers) has provided greater choice and flexibility for DRC members. We expect DRC member usage of the core to remain steady and strong, given attractive new technologies afforded by spectral flow cytometry. CCSC will assist investigators in two ways: Aim 1 – To quantify and phenotypically characterize cell populations that contribute to the metabolic, immunological, and developmental programs of diabetes and its complications. CCSC will leverage advanced technologies in fluorescent imaging at the single cell resolution to support the analysis of different cell types in human and animal tissues, including humanized mouse models of Type 1 diabetes. Aim 2 – To purify populations of cells of relevance to diabetes and its complications. Using fluorescence-activated cell sorting, the CCSC will support DRC investigators to purify individual populations of cells for culture, in vivo implantation, or molecular characterization including genetic and transcriptional profiling, protein purification and signaling studies, and for functional analysis including T cell activation, proliferation, and migration studies. To achieve these aims, the CCSC has established standard operating procedures to: (i) assure quality control and reproducibility; (ii) prioritize investigator use; (iii) monitor Core usage; and (iv) adapt to new technologies and to the needs of the Columbia Research Base.