4: Sample Experiments
► Procedure
Follow the steps below to take an absorbance measurement using OOIBase32:
1. Place OOIBase32 in scope mode by either clicking the scope mode icon on the toolbar, or
selecting Spectrum | Scope Mode from the menu.
2. Ensure that the signal is on scale. The peak intensity of the reference signal should be about
50,000 counts.
3. Ensure that nothing is blocking the light path from the sample to the spectrometer, and then take a
reference spectrum. You must take a reference spectrum before calculating absorbance.
Note
The material to measure must be absent while taking a reference spectrum.
4. Click the Store Reference spectrum icon on the toolbar or selecting Spectrum | Store Reference
from the menu to take the reference. This command merely stores a reference spectrum. You
must select File | Save | Reference from the menu to permanently save the spectrum to disk.
5. Block the light path to the sample and 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. Storing a dark spectrum is required before the software
can calculate absorbance spectra.
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. If you must turn off your light source to store
a dark spectrum, make sure to allow enough time for the lamp to warm up before
continuing your experiment.
6. Ensure that the sample is in place and the light path is clear, and then take an absorbance
measurement. Click the Absorbance Mode icon on the toolbar or select Spectrum | Absorbance
Mode from the menu. To save the spectrum, click the save icon on the toolbar or select File |
Save | Processed from the menu.
Note
If at any time any sampling variable changes (including integration time, averaging,
boxcar smoothing, etc.), you must store a new reference and dark spectrum.
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