Bio-Rad Bio-Plex Pro™ Human Cancer Biomarker 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 (pg/ml). 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