horiba FluoroMax-3, Operation Manual, Page: 66 / 220

FluoroMax-3 v. 3.1 (3 Mar 2006) Data Acquisition
4-14
e
Set the scan parameters.
Use the emission maximum determined above for the excitation, use 250 nm for
starting, enter the emission maximum minus 15 nm for the end of the scan, and
select two acquisition modes,
S
and
S/R
.
S
collects raw signal from the emis-
sion detector, and
S/R
ratioes the signal to the reference detector.
f
Set excitation and emission slits identical to the emission scan.
Be sure that the emission scan did not exceed 20 µA in the emission scan.
g
Enter the
Experiment File
name.
h
Click Run .
The scan starts.
The resulting spectrum shows maximum excitation wavelength. If the raw S-
channel signal > 20 µA, reduce slits, and rescan.
i
Note the excitation peak.
This is the optimum excitation peak position.
3
Find the optimal emission peak.
a
Use the optimum excitation wavelength determined in step 2.
b
When complete, both excitation and emission peaks are found.
Optimized excitation and emission spectra of a 1 × 10
–8
- M anthracene solution
are shown below. Because the acquisition modes were different for the excita-
tion and emission scans, the data intensity had to be normalized. After normali-
zation, the excitation and emission scans are virtually mirror images of one an-
other.
300 350 400 450 500
Wavelength (nm)
Intensity (1
0
6
counts s
–1
)
4
3
2
1
0
Excitation
Emission
Normalized excitation and emission spectra of a 1 × 10
–8
-M
anthracene solution.