Flow & Imaging Cytometry Core Facility

National Institute Of Neurological Disorders And Stroke

Investigators

Linked publications & trials

Abstract

NINDS Flow and Imaging Cytometry Core Facility (Facility) was established in 2001 by NINDS DIR as a crucial resource to NINDS and other intramural investigators to support their work in both basic and clinical research programs. The Facility continuously provides seamless use of flow cytometry and imaging equipment managed by the Facility as well as the crucial expertise to train investigators in proper and optimal use of this equipment. The Facility staff is also available to provide constructive input in designing and optimization of a variety of novel and customized multiplex staining protocols using unique combinations of appropriate cell surface, cytoplasmic, nuclear, protein, gene expression and physiological indicator dye biomarkers that are of specific interest to initiating investigator(s) for any given project. When requested, the Facility staff routinely carry out multi-color flow and in situ cytometry experiments and analyze the data from single or multiple cell populations of interest. Applying user-defined Boolean logic gating strategies based on different combinations of biomarkers tested, the Facility provides quantitative accounts and interpretations of the complex signal distributions in individual cells or cell populations of interest to each investigator. Using the preparative cell sorting capability of flow cytometers, the Facility staff is available to assists with isolation of highly purified cell populations and subpopulations from heterogeneous cell preparations, as requested by the initiating investigator. These include sorting of neural stem and progenitor cells, specific neuronal and glial cell types, as well as non-neural cell types (microglia, endothelial cells) from both CNS and PNS tissues. In addition, the Facility routinely sorts non-CNS-derived stem cells (hematopoietic, mesenchymal and Hoechst dye-excluding side populations) and different leukocyte phenotypes (lymphocytes, monocytes, neutrophils, basophils, eosinophils, mast cells) from the bone marrow, peripheral blood, spleen, and cerebrospinal fluid. These highly-purified cells of interest are then routinely used by NINDS and other intramural investigators for further in vitro and in vivo studies including cell culture, transplantation, time-lapse and physiological indicator dye imaging, and for transduction or transfection with specific gene constructs of interest. Additionally, FACS-purified cells have been utilized as an invaluable source of proteins for Western blotting and immunoprecipitation experiments as well as a source of RNA for RT-PCR, microarray, RNA-Seq, and microRNA assays. These cells have also provided unprecedented insights into specific loss-of-function experiments using antisense and siRNA probes. The major significance in using FACS-purified homogeneous cells in the abovementioned applications is that this approach reduces the ambiguity in the findings inherently present when studying heterogeneous cell preparations. Taken together, the routine applications of quantitative flow and in situ cytometry assays and preparative cell sorting services provided by this Facility has produced numerous insights into cell biology for NINDS and other intramural investigators, which could not otherwise be demonstrated. Since inception in 2001, the Facility has provided these invaluable services to more than 500 NINDS and other intramural investigators, fellows, students, and scientific staff and supported the multidisciplinary scientific research in over 700 basic, translational, and clinical research projects from 105 laboratories across 15 NIH institutes and centers, culminating in publication of over 240 peer-reviewed articles in highly acclaimed biomedical and biological scientific journals. Over the past 12 months, the Facility supported the research needs of 112 investigators, fellows, students, and laboratory staff contributing to 104 basic science, clinical and translational research projects in 28 NINDS and other intramural laboratories and facilitated multidisciplinary collaborative projects between NINDS and 6 other NIH institutes and centers. Contributions by this Facility over the past 12 months were incorporated in publications of 11 peer-reviewed articles in highly acclaimed scientific journals and expedited the preparation of 15 more articles currently in preparation for journal submission or in journal peer review.