Eppendorf BioSpectrometer fluorescence - Manual (Page 36)

Methods
Eppendorf BioSpectrometer
® fluorescence
English (EN)
36
Calibration curve
• Measurement of the RFU values and evaluation with a series of 2 to 12 standards
• Various evaluation procedures, including linear regression ("Curve fit") and nonlinear regression, can
be selected.
• Graphical and tabular display of the standard results.
• The last saved standard evaluation can be used.
• A method for calibration curve evaluation is preprogrammed.
6.4 Method parameters
This chapter illustrates the parameters for programming the methods. The order of the parameters in the
device display may slightly differ from the order in the table in order to display the parameters more clearly.
The table displays all parameters available for the various methods. Only a small portion of these
parameters are required for the corresponding method and will be shown in the display.
Parameter Entry Explanation
Cuvette Selection:
10 | 5 | 2 | 1 | 0.5 | 0.2 |
0.1 mm Optical path length of the cuvette. The device always
automatically converts absorbance values to the 10 mm path
length of a standard cuvette (see Absorbance values on p. 87).
Therefore, there is no need to change factors such as "50" for
the calculation of dsDNA concentrations when modifying the
Cuvette parameter.
No. of
wavelengths Value input:
Range: 2 to 6. Only for the Multi λ method group.
Number of wavelengths at which the measurement is to be
performed.
Wavelength Value input:
Measurement
wavelength in nm.
Range: 200 to 830 nm. Measurement wavelength: The concentration is calculated
based on the absorbance measured with this wavelength.
For the
Multi λ and Dual wavelength method groups, enter
more than one wavelength. For some method groups (e.g.,
Nucleic acids and Proteins direct UV), the wavelengths are
preprogrammed.
For the
Dye labels method group the measuring wavelengths
are not entered into the method procedure one by one. They
are automatically imported simply by selecting the biomolecule
and dye from the
General Method Parameters function.
Wavelength
(em) Selection:
520 nm | 560 nm Only for the Basic fluorimetric method group:
Measurement wavelength: The concentration is calculated
based on the fluorescence measured with this wavelength
(emission wavelength).
For the methods of the
Routine group the wavelength is
preprogrammed.
Wavelength (ex) No entry possible.
Wavelength: 470 nm
Only for fluorimetric method groups:
Excitation wavelength 470 nm is displayed.