Bio-Rad Bio-Plex Pro™ Human Isotyping Assays - Manual (Page 5)

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Fig. 1. Bio-Plex sandwich immunoassay.
Data Acquisition and Analysis
Data from the reactions are acquired using a Bio-Plex system or similar
Luminex-based reader. When a multiplex assay suspension is drawn into
the Bio-Plex 200 reader, for example, a red (635 nm) laser illuminates the
fluorescent dyes within each bead to provide bead classification and thus
assay identification. At the same time, a green (532 nm) laser excites PE
to generate a reporter signal, which is detected by a photomultiplier tube
(PMT). A high-speed digital processor manages data output, and Bio-Plex
Manager
™ software presents data as median fluorescence intensity (MFI) as
well as concentration. The concentration of analyte bound to each bead is
proportional to the MFI of the reporter signal.
Biomarker
of Interest
Magnetic Bead
Capture
Antibody
Biotinylated
Detection
Antibody
Phycoerythrin
Fluorescent
Reporter
Streptavidin