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- W289989476 abstract "The Flow Cytometry Research Group (FCRG) is the latest addition to the ABRF RG family. The RG is currently in its first year and has 9 members; many of whom are flow cytometrists new to the ABRF. The initial goal of the FCRG is to describe a method for the evaluation of cell stress or other deleterious perturbations caused by cell sorting across a wide range of cell types.As an example, reports have been published demonstrating the susceptibility of dendritic cells (DCs) to phenotypic and functional changes after manipulation and isolation using techniques such as magnetic bead enrichment. To evaluate the effects of cell sorting on DCs, populations were enriched on both jet-in-air (MoFlo and FACSVantage ) and cuvette-based (FACSAria II) cell sorters, and then assessed in vitro for their ability to function as antigen presenting cells. This study showed that DC populations sorted on a cuvette sorter had decreased numbers of viable cells and decrease ability to induce antigen specific T cell proliferation, while cells sorted on jet-in-air sorters were able to induce proliferation and deemed functional.The FCRG is expanding upon this work with a 3-year research plan study which will utilize microarray analysis as an aid to identify optimal cell sorting conditions for a wide variety of cell types. This year's pilot study utilized Jurkat cells sorted through a Beckman Coulter MoFlo Legacy flow cytometer with two nozzle sizes at their respective pressures ( 50um nozzle at 70psi compared to 100um nozzle at 20psi), with and without UV laser exposure. Cells were sorted and allowed to incubate for 3 hr in RPMI media in parallel with a matched unsorted cellular control. Total RNA was extracted using Trizol and analyzed using the new Affymetrix Human Gene ST 2.0 microarrays. Results of this pilot study will be presented." @default.
- W289989476 created "2016-06-24" @default.
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- W289989476 date "2013-05-01" @default.
- W289989476 modified "2023-09-27" @default.
- W289989476 title "Overview of the New Flow Cytometry RG and Proposed Cell Sorting (FACS) Microarray study" @default.
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