Eppendorf BioSpectrometer fluorescence - Manual (Page 96)

Evaluation procedure
Eppendorf BioSpectrometer
® fluorescence
English (EN)
96
12.8 Fluorimetry
12.8.1 RFU values
Relative Fluorescence Unit: RFU values are a measure of the measured fluorescence. Unlike the
absorbance values in photometry, the RFU values are not comparable from device to device, therefore they
always need to relate to known concentration or fluorescence standards.
12.8.2 Blank
All RFU values always relate to the last measured blank (value).
Therefore, a blank measurement is compulsory at the start of every series of measurements and can be
completed at any time during a series of measurements. Ideally, the blank measurement should be able to
compensate for any influences on the RFU value of the measuring solution. The blank should therefore be
measured with the same buffer that was used for the sample measurement and the same cuvette that was
used to measure the sample value – unless the cuvettes used for the blank and sample measurements are
optically aligned and thus have the same RFU value at the measuring wavelength.
12.8.3 Evaluation with standard and standard curve/line, dilution
The evaluation with a standard or with standard curve/line is similar to the evaluation of photometric
methods (see Evaluation with factor or standard on p. 88).
If evaluations are made with more than one standard, different evaluation procedures for the standard
curve/line can be selected with the [Curve fit] in the
measure standards/new method step (see Evaluation
with standard curve/line on p. 89).
The way the results of the dilution is calculated is identical to that of photometric methods (see Dilution on
p. 90).