Luminex 100 IS Version 2.2 - Manual (Page 11)

x MAP Technology General Guidelines
PN 89-00002-00-032 Rev. A 5
Bead Concentration
The concentration of beads in an assay determines system speed. To
analyze a 96-well plate in one hour, we recommend you use at least
5000 beads per target and do not exceed a bead concentration of 2.5
x 10
6
microspheres/mL.
Repetitive Microsphere
Measurements
The reporter, bound to reagents at the surface of the microspheres,
provides raw analytical data. Because a microsphere suspension
provides near liquid phase reaction kinetics, each microsphere of a
particular signal theoretically binds an equal number of reporter
molecules. Equal binding results in a statistically even distribution of
reporter on each microsphere in a set. Therefore, when the Luminex
100 IS system registers numerous events for each set, the system
reports individual population values based on an even coating of
reporter. This means that numerous replicates for each microsphere
population are measured from a single reaction vessel. The
confidence in a given measurement strengthens with increased
replicate measurement. The median value for the microspheres
having a specific signal is reported as the result.
Classification and
Reporter Fluorochromes
Each microsphere is dyed to emit light in a certain classification
channel. All microspheres of a given emission represent a distinct
assay within a multiplex of assays. A reporter channel is used to
detect fluorescence bound to the surface of each microsphere, and
each reporter emission quantitates each of the distinct assays.
Table 1: Bead Selection
033 034 035 036 037 038 042 043 044 045
046 047 051 052 053 054 055 056 061 062
063 064 065 066 072 073 074 075 076 077
017 018 019 020 021 024 025 026 027 028
029 032 041 011 012 013 014 006 007 008
083 084 085 086 087 088 089 090 048 057
058 067 068 069 078 079 080 002 003 004
009 015 022 030 039 049 050 059 060 070
071 081 082 001 005 010 016 023 031 040
091 092 093 094 095 096 097 098 099 100