Molecular Devices FlexStation II, Руководство оператора

Страница: 87 / 134

3.10 Optimizing Fluorescence Assays in Spectrum Mode
(Follow the plot to the left of the peak until the RFU value falls to approximately
90% of the maximum and then drop a line from the 90% point on the plot to the x-
axis – refer to the figure below.)
RFU
Wavelength
RFU at 90% of max
λ
max
λ
90% of max
λ
Plot of RFU vs. Wavelength
2 Preliminary Emission Scan (Scan Figure 2)
The preliminary emission scan without filters allows you to see the actual emission
spectrum peak.
a
In SOFTmax PRO, set up a second Plate section for a fluorescence read, spectrum
mode, Ex Fixed/Em Scan, with no cutoff filter (default), and medium PMT.
b
Set excitation wavelength to the value determined in
1f
above.
c
Set emission scan to start/stop approximately 50 nm below/above the tentative
emission value obtained from the literature (or existing filter pair).
d
Set step increment to 1–2 nm (or do a preliminary scan with a 10-nm increment to
determine the approximate peak location and then repeat the scan over a narrower
wavelength range using a 1–2 nm increment.)
e
Perform scan and view results as a plot of fluorescence vs. emission wavelength.
Scan Figure 2
FlexStation II Operator’s Manual – Rev. D 81