Bio-Rad Bio-Plex Software® Upgrades and Conversions - Manual (Page 240)

Bio-Plex Manager Software 6.1 User Guide | Appendix
228
Fluidics
There are two fluidic paths in the array reader. The first path includes a
syringe-driven mechanism that controls the sample uptake. This mechanism
permits small sample uptake volumes from small reaction volumes. The
syringe-driven system transports a user-specified volume of sample from a
microplate well to the cuvette. The sample is injected into the cuvette at a
steady rate, and the microspheres in the sample are aligned in a single file
through the path of the two lasers.
Following analysis, the first sample path is automatically purged with sheath
buffer fluid via the second fluidics path. This process effectively removes
residual sample in the tubing, valves, and sample needle.
Excitation
There are two lasers in the array reader: one for classifying each microsphere
and its associated analyte, and the other for quantitating the amount of
analyte bound to each microsphere. The first laser, the classification or "red"
laser, excites the dyes embedded in each microsphere; the fluorescent signal
is discriminated with selective emission filters and converted into intensity
units by fluorescence detectors and a digital signal processor, and the
microsphere is classified. The second laser, the reporter or "green" laser,
excites the fluorochromes bound to the target analytes on the surface of the
microspheres; again, the fluorescent signal is discriminated with selective
emission filters and converted into intensity units by fluorescence detectors
and a digital signal processor, and the amount of analyte is quantitated.
BioPlex_6.book Page 228 Friday, September 23, 2011 11:55 AM