Ocean Optics OOIBase32 - Manual (Page 100)

A: Experiment Tutorials
2. Ensure that the entire signal is on scale. The intensity of the reference signal should peak at about
3500 counts. If necessary, adjust the integration time until the intensity is approximately 3500
counts.
3. Place a sample of the solvent into a cuvette and take a reference spectrum. You must take a
reference spectrum before measuring absorbance.
Note
Do not put the sample itself in the path when taking a reference spectrum, only the
solvent.
Click the Store Reference spectrum icon on the toolbar or select Spectrum | Store Reference
from the menu bar to store the reference. This command merely stores a reference spectrum in
memory. You must select File | Save | Reference from the menu bar to permanently save the
spectrum to disk.
4. Block the light path to the spectrometer. Then, take a dark spectrum by clicking the Store Dark
Spectrum icon on the toolbar or by selecting Spectrum | Store Dark from the menu bar. This
command merely stores a dark spectrum in memory. You must select File | Save | Dark from the
menu to permanently save the spectrum to disk.
Note
If possible, do not turn off the light source when taking a dark spectrum. If you must turn
off your light source to store a dark spectrum, allow enough time for the lamp to warm up
again before continuing your experiment. After the lamp warms up again, store a new
reference (Step 3).
You must take a dark spectrum before measuring absorbance.
5. Put the sample in place and ensure that the light path is clear. Then, take an absorbance
measurement by clicking on the Absorbance Mode icon on the toolbar or selecting Spectrum |
Absorbance Mode from the menu. To permanently save the spectrum to disk, click the Save
icon on the toolbar or select File | Save | Processed from the menu bar.
Note
If you change any sampling variable (integration time, averaging, smoothing, fiber size,
etc.), you must store a new dark and reference spectrum.
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