Luminex 100 IS Version 2.2 - Manual (Page 12)

Luminex 100 IS Developer Guide to x MAP Technology Version 2.2 x MAP Technology
6 PN 89-00002-00-032 Rev. A
Table 2 displays acceptable reporter fluorochromes wavelengths
excitation/emission.
Sample Size
Sampling volume or sample size can range from 20 to 200
microliters. Assays must have enough volume in the wells so that
after this sampling volume is taken up, some sample remains. Make
the volume in the wells at least 25 uL greater than the sample
volume. This amount may vary depending on the type of plate used.
After acquisition, the instrument washes the sample lines resulting in
ejection of approximately 160 microliters of sheath fluid back into
the well. Make sure that there is room to add this amount to the well
without overflowing the well and contaminating other wells.
Note: The volume restrictions that the amount expelled back
function places on the assay design can be expounded by the
following formula.
Table 2: Reporter Fluorochromes
R-
Phycoerythrin
Alexa
532
Formula weight (Daltons) 240,000 470
Absorbance max (nm) 480, 546, 565 531
Extinction max (M -1 cm -1 )
1,960,000 83,800
Emission max (nm) 578 554
Quantum yield 0.82 0.8
[“Total well volume” (uL ) – “Sample uptake volume”(uL)] + 160(uL)
Where:
Total well volume The starting sample volume of a well
before the unit samples for acquisition.
Sample uptake
volume The volume the unit uptakes for
acquisition (program this in the template
as sample vol.)
160uL The amount expelled back volume stated
in this section.
Maximum Well
volume plate The maximum volume capacity of the
user’s selected 96 well microtiter plate.