Fluorolog-3 v. 2.2 (31 Jul 2002)
Producing Correction Factors
11-15
Calculating excitation correction factors
The photodiode reference detector handles the bulk of excitation correction from 240–
600 nm when a ratio acquisition mode is selected (e.g.,
S
/
R
for single-beam and T-box
sampling modules). More accurate measurements require that compensation be applied
for the difference in optical path between the detector and the sample. This can be ac-
complished by a simple excitation scan with rhodamine-B placed in the sample posi-
tion.
1
Fill a cuvette with a solution of rhodamine-B.
Use 8 g L
–1
of laser-grade rhodamine-B in 1,2-propanediol.
2
Place the cuvette in the sample compartment.
For single-beam sampling modules, place the cuvette in the standard cell holder
and select right-angle detection.
3
Enter the
Real
Time Display
.
4
Set the excitation
and emission
monochromators
to 467 nm and 630
nm, respectively.
The largest lamp peak occurs
at 467 nm.
5
Set the two slits
on the excitation
spectrometer to
0.5 mm.
6
Make sure the
shutter is open.
7
Set the excitation monochromator to 560 nm
and the emission monochromator to 630 nm.
8
Adjust the slits on the emission monochromator.
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