Application Note

Analysis Of Mixed Microbial Populations And Deep Immunotyping Of Peripheral Blood On The ZE5™ Cell Analyzer

Source: Bio-Rad Laboratories, Inc.

Angie Green PhD, Kristin Lind Ph.D., Michelle Scott, and Kelly Kroeger Ph.D.

Red Blood Cells

With up to five lasers and 30 analysis parameters, the ZE5 Cell Analyzer is well equipped for complex multiplex
analysis of cells. Excellent fluorescence sensitivity (<100 MESF values in FITC, PE, and APC channels) enables detection of antigens expressed at low levels. The three scatter options — forward scatter (FSC), side scatter (SSC), and forward scatter small particle detector (FSCspd) — provide optimal resolution of particles >0.3 to 50 μm in size. Furthermore, the option to set a dual trigger using any scatter or fluorescence channels allows for optimal background reduction.

The ZE5 Cell Analyzer was used to successfully perform immunophenotyping of multiple cellular subsets in human blood using multicolor panels. Peripheral blood mononuclear cells (PBMCs) from normal healthy donors were stained using a 19-color panel to identify T cell, B cell, NK cell, dendritic cell, and monocyte subpopulations. Results showed that the panel data were compensated accurately on the ZE5 Cell Analyzer and that each cellular subpopulation was well identified.

The ZE5 Cell Analyzer also effectively differentiated size beads and mixed microbial populations. Beads sized ≥0.2 μm were discriminated from noise when using scatter or fluorescence as the trigger. Analysis of mixed microbial populations clearly identified the unique populations within the mixture.