BD Influx

Unique properties revealed with polarization-sensitive detectors

Unique properties revealed with polarization-sensitive detectors

A. Viewed with polarization-sensitive detectors on the BD Influx™ 200-mW, 488-nm laser, liths (scales) and lithed cells of the phytoplankton E. huxleyi can be discriminated using perpendicular and parallel FSC.

B. Polystyrene beads serve as a negative control.

Prochlorococcus and polystyrene bead scatter using the Small Particle Option
Prochlorococcus and polystyrene bead scatter using the Small Particle Option

A. Small cells of the cyanobacteria Prochloroccocus are visible when acquired with the BD Influx system with the Small Particle Option.

B. The 100-nm polystyrene beads would not be visible without the Small Particle Option.

Hierarchical gating strategy

Hierarchical gating strategy

Human lymphocyte subpopulations were identified with fluorochrome-conjugated antibodies excited by the 488-nm, 561-nm, 640-nm, and 405-nm lasers. A hierarchical gating strategy identified peripheral blood mononuclear cells (PBMCs) (forward scatter vs side scatter), singlets (forward scatter vs forward scatter width), lymphocytes (CD45-PerCP-Cy™5.5 vs side scatter) and lymphocyte subset populations (fluorescence plots).

Four-way sorting

Four-way sorting

The BD Influx system was set up for a 4-way sort using an 86-μm nozzle (30 psi with a frequency of 60 kHz). Lymphocyte subset populations were sorted in purity mode at a rate of 10,000 events per second. Sort purity was 98% or greater.


 

Real-time viewing

Real-time viewing

Real-time viewing of the sort in process is enabled by live videos of the side streams, drop breakoff point, and laser intercept on the stream.

Density plots: Density plots show two variables plotted against each other using color to illustrate heavily concentrated events.

Contour plots: Like density plots, contour plots show two variables plotted against each other, but instead use topographic lines to show the boundaries of populations.

Analysis of contour plot: Employs topographic lines to define subpopulations or move the cursor over a topographic line for instantaneous subpopulation statistics.

Discrimination of small particles can be further improved by measuring fluorescence

Discrimination of small particles can be further improved by measuring fluorescence

Representative Hoechst staining of Pelagibacter ubique.


 
Analysis of aquatic samples

Analysis of aquatic samples

Aquatic samples contain a large variety of very small particles. Data shows forward scatter (200 mW, 488 nm) vs fluorescence (692/40) in samples taken at different depths from the Sargasso Sea. Distinction of Prochlorococcus and Synechococcus cyanobacterium clusters by flow cytometry illustrates the exceptional sensitivity of the standard BD Influx forward scatter detector.

Measurement of macrophage cell line stimulation using GFP

Measurement of macrophage cell line stimulation using GFP

Murine RAW macrophage cells were transfected with a Green Fluorescent Protein (GFP) and stimulated with varying concentrations of the TLR ligand PAM2. Degradation of GFP fluorescence was measured over a 50-minute time period using a BD Influx system equipped with a 200-mW 488-nm laser. The data was binned and averaged to produce the mean fluorescence for each time point. The BD Influx system is able to make many measurements over short time scales to increase the accuracy of time course studies.

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