SpectraMax L Luminescence Microplate Reader User Guide
29
0112-0174 C
9. Select the
Sensitivity
category.
a. Click the
PMT Settings
drop-down:
Select
Auto Range
to combine the benefit of both Analog Only (at a higher
range) and Photon Counting modes. PMT signals are simultaneously
converted to both analog current and digital pulses to achieve high
sensitivity and wide dynamic range.
Select
Analog Only
for very bright signals above 1.6 x 10
7
RLU. PMT voltage is
set low, and the measurement is based on current.
Select
Photon Counting
for very dim and medium signals below 25 x 10
6
RLU.
This allows discrimination between the signal and the noise generated in the
dynode chain. PMT signals are converted into digital pulses, which can be
measured above a set threshold.
Select
Max Range
to combine Auto Range and Analog Only to maximize the
dynamic range (10 to 1 x 10
9
RLU). Detection occurs in Auto Range mode until
the crossover point, then switches to Max Range mode with its low PMT
voltage setting when samples are extremely bright. Select for assays with
massive dynamic range.
b. If you select Auto Range, Analog Only, or Max Range in the previous step, you
must select the
Target Calibration Wavelength
option that most closely
corresponds to the peak emission wavelength of your assay:
Note:
The sensitivity of the photomultiplier tube (PMT) depends on the
wavelength of the light. The instrument is calibrated at several
wavelengths. Select the Target Calibration Wavelength that is close to your
sample's emission wavelength for best RLU comparability. Note that if you
want only specific wavelengths to be detected, you must install optical
filters. See
Installing the Filters on page 19
.
Select
395 nm (Acridinium Ester, DeepBlueC)
Select
470 nm (Dual Firefly/Renilla Luciferase, Renilla Luciferase, CSPS,
CDPstar, Aequorin, Vibrio Luciferase, AMPPD, Luminol, Coelentrazine
Select
527 nm (Lumi Phos 530)
Select
570 nm (Firefly Luciferase)
c. If you select Photon Counting, select a
Correction Option
:
Select
No Correction
if you do not want to apply a custom calibration.
Select
Custom Calibration
and click the
Custom Calibration #
drop-down
and select
1
or
2
. One and two correspond to the user-defined custom
calibrations that you define on the Calibration dialog. See
