![]() Briefly, cell products were washed, and resuspended in PBS-EDTA (2.5. The function fitting algorithm applied by FacsKin is suitable to provide a common basis for evaluating and comparing flow cytometry kinetic data.Ĭopyright © 2013 International Society for Advancement of Cytometry. Flow cytometry was performed on a MACSQuant. With the algorithm of function fitting instead of smoothing, more statistically significant differences of potassium channel inhibition between the two RA groups could be demonstrated. Our skilled staff provide a variety of flow cytometric approaches for. While initially Th1 cells are less sensitive to the inhibition of Kv1.3 and IKCa1 channels in RA, their sensitivity increases along with the duration of the disease. The University of Kentucky Flow Cytometry and Immune Monitoring Core Facility offers state-of-the-art flow cytometry cell analysis, high-speed sorting, and immune functional analysis to the research community at UK, and has been in continuous operation since 1983. The facility also has a site license for FlowJo. Th2 cells of patients with established RA react slower to activating stimuli, whereas CD8 cells show a faster reaction than in patients with recently diagnosed RA. The workstations in the lab have either Mac-based CellQuest analytical software or PC-based Summit software. We assessed the sensitivity of the above subsets to specific inhibition of the Kv1.3 and IKCa1 potassium channels. Flow Cytometry Core Influxes (Jet-in-Air technology) FACSAria Fusion (Fixed alignment cuvette) BD FACS Symphony A3 LSRII-1 and LSRII-2 Canto II MACSQuant. We evaluated calcium influx kinetics following activation in CD4, Th1, Th2, and CD8 cells applying an approach based on smoothing of median fluorescence values (FlowJo) and an algorithm based on function fitting (FacsKin). Commercially Available Software FlowJo FlowJo CellEngine CellCarta FCS Express 7 - DeNovo Software NovoExpress - ACEA Biosciences WinList - Verity. We took peripheral blood samples from nine patients with recently diagnosed and six patients with established RA. We aimed to compare peripheral T lymphocyte calcium influx kinetics upon activation in patients with recently diagnosed and established RA, and to demonstrate the differences in analysis of kinetic flow cytometry data when using two different algorithms. Kv1.3 and IKCa1 potassium channels are important regulators of the maintenance of calcium influx during lymphocyte activation and present a possible target for selective immunomodulation. ![]() The transient increase of the cytoplasmic free calcium level in T lymphocytes plays a key role in initiating and maintaining the autoimmune reaction in rheumatoid arthritis (RA). ![]()
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