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Signals were amplified with the logarithmic mode for side scattering, forward scattering, and FITC fluorescence.
Samples were analysed on a CyAn ADP high-performance flow cytometer (Becton Dickinson, Oxford, UK); cells were gated according to size scattering, forward scattering and pulse width so only single cells were analysed.
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The cell sizes were inversely proportional to the amount of light scattered forward and at right angles.
Side scatter, forward scatter, and pulse-width gates excluded debris and aggregates.
Inflammatory cell populations (monocytes, neutrophils, eosinophils, and lymphocytes) in blood were identified and gated based on light scatter properties (forward scatter and side scatter).
Side scatter versus forward scatter dot plots were used to locate and gate nuclear populations by particle size.
Cells were gated in a SSC (side scatter)/FSC (forward scatter) plot with the scatter gate for monocytes partially extending into lymphocytes [ 25].
The dot plots in Fig. 3A show differences in side scatter and forward scatter profiles between unprimed and primed cells cultured with infected PL's for 72hrs.
The following parameters were used: FL1 for Annexin V, FL2 for PI, side scatter and forward scatter.
DC were gated based on side scatter and forward scatter followed by doublet discrimination gating (Fig. 1A).
The subpopulations with high fluorescence had similar side scatter and forward scatter profiles to the whole population indicating that the cells were intact.
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