FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
112 0112-0127 B
Optimizing Luminescence Assays
Luminescence can be read from the top or the bottom of a microplate.
Solid white plates or white plates with clear bottoms are recommended
for luminescence reads.
For standard luminescence, a separate light path without
monochromators carries the emitted light to a dedicated PMT. The
optimum emission wavelength is between 360 nm and 630 nm. Under
reader setup the emission says All.
For wavelength-selectable luminescence, the emission monochromator
is used to differentiate the wavelengths being emitted from the well. Up
to four emission wavelengths between 250 nm and 850 nm can be
specified. If reading only one luminescent event in the well, best
sensitivity should be achieved using the standard luminescence
measurement, without a wavelength selected.
Luminescence read times are not designated by multiple reads per well,
but rather by choosing the total integration time desired between 1 ms
and 1500 ms. Typical luminescence assays require between 500 ms and
1000 ms integration.
If wells have been incubating for a long period of time, it is a good idea to
mix the plate before reading. This can be done using Automix in the
reader.
If it appears that the signal is always higher in the first wells read (such
as, column A), the plate might need to be “dark adapted” to reduce the
auto-luminescence of the white plastic. The auto-luminescence
decreases quickly, so manually load the plate from the control panel and
wait for 1 to 2 minutes before initiating the read and then determine if
the read-out is more consistent across the plate.
FlexStation_manual.book Page 112 Tuesday, June 25, 2013 10:22 AM
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