Bio-Rad VersaFluor™ Fluorometer - Manual (Page 19)

5.2 Instrument Parameter Set-Up
1. The VersaFluor Fluorometer instrument must warm up for 20 minutes prior to use. If the
instrument is not fully warmed up, the reading may be irreproducible.
2. Select an excitation and emission filter which are to be inserted in the filter holders. Orient
the filters so that the notched groove on the filter holder faces the cuvette holder.
Caution: Use filters which are tailored to the sample being measured. Using incorrect
filters will greatly degrade the accuracy of the fluorescence measurement.
3. Press the SETUP button on the main menu to access the print averaging and gain set-
tings.
4. Press the AVRG to select the print averaging time (1, 2, or 4 seconds).
5. Press GAIN to select the gain setting (LOW, MED, or HIGH). Then press EXIT to return
to the main menu.
6. Add the standards, unknown samples, and blanks to the disposable cuvettes.
7. Follow the sample measurement guidelines below to insure accurate results:
• Warm up the instrument for at least 20 minutes.
• Always use calibrated pipets to insure accurate pipetting.
• Mix standards and sample completely by using a disposable transfer pipet.
• Remove any air bubbles in the cuvettes.
• Hold fluorometer cuvettes by the upper edges since the cuvettes have four optically
clear sides.
• Clean the cuvette sides with a lint-free tissue.
• Read all standards and samples at ambient temperature.
• While taking a reading, keep the samples in the fluorometer only. This helps reduce
sample photobleaching.
5.3 Setting Up a Standard Curve
The range should be set on the fluorometer with a known standard and a fluorescent dye.
Depending on the protocol used, a single-point or multi-point standard curve is required.
1. Set the range of the instrument to 00000 by pressing the RANGE button. Adjust the range
to 00000. Press EXIT.
2. Zero the instrument by placing the baseline cuvette (blank) in the cuvette holder and clos-
ing sample compartment lid. Wait approximately 10 seconds for the detector to adjust to
the light conditions. Press the SET ZERO button; when blinking stops, read the fluores-
cence display. If fluorescence display is 0 ± 5, the instrument is zeroed. If not, re-zero
the instrument by pressing the SET ZERO button.
3. Set the range of the instrument by placing the highest concentration standard cuvette in
the cuvette holder and closing the sample compartment lid. Press the RANGE button.
Adjust the range to the desired setting. The maximum setting is 19,999 RFU. Press EXIT.
The fluorescence display should read the set range value. Read or print the RFU number
in the fluorescence display. Remove the highest concentration standard cuvette.
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