The lower limit of the lag time was chosen to exclude scattered waves near the receiver, and the upper limit corresponds to the time at which the amplitudes of the ACFs converged to their noise levels.
Exposure of specimens to the film was conducted in a lead-shielded surrounding to exclude scattered radiation at −20°C.
Moreover, owing to the short life span of normal granulocytes, the purity of the MM cell populations increased over culture days and, eventually, the cultures became homotypic, excluding scattered macrophages (Supplementary Figure 1).
The blocking bar, employed in all flow cytometers to exclude direct laser exposure and reflection into the detectors, also excludes light scattered below a certain critical angle.
The cells were threshold on Forward Scatter (log amplified) to exclude dust particles.
The small active area (∼180 μm in diameter) of the silicon APD detector serves as a limiting aperture to exclude laser light scattered from the cuvette windows and the sheath fluid.
Dead cells and debris were excluded by forward scatter, side scatter, and Amine Aqua Viability Dye measurement.
For all analyses, dead cells and debris were excluded by forward scatter (FSC), side scatter (SSC) and amine aqua viability dye.
Doublets were excluded using side-scatter height (SSC-H) vs side-scatter width (SSC-W) and forward-scatter height (FSC-H) vs forwardscatter width (FSC-W) parameters.
Cells were analysed using the LSR Fortessa (BD Biosciences) and for all cell subsets doublets were excluded by forward/side scatter width compared with height and side scatter-width compared with height and only CD45+ events included in the analysis.
