Molecular Devices SpectraMax Paradigm User Manual page 30

SpectraMax Paradigm Multi-Mode Detection Platform User Guide
The instrument permits scanning of both excitation and emission wavelengths, using
separate tunable dual monochromators. One benefit of scanning emission spectra is that
you can determine more accurately whether the emission is, in fact, the expected
fluorophore, or multiple fluorophores, and not one generated by a variety of background
sources or by contaminants. One more benefit is that you can find excitation and emission
wavelengths that prevent interference when interfering fluorescent species are present.
For this reason, it is desirable to scan emission for both an intermediate concentration of
labeled sample, as well as the background of unlabeled sample. The optimum setting is
where the ratio of the sample emission to background emission is at the maximum.
Fluorescence intensity data is dependent on a number of variables. See
Fluorescence Intensity Data on page
Applications of Fluorescence Intensity
Fluorescence intensity is used widely in applications such as fluorescent ELISAs, protein
assays, nucleic acid quantitation, reporter gene assays, cell viability, cell proliferation, and
cytotoxicity. One more major application of this mode is to study the kinetics of ion release.
Some assays use a fluorescent label to selectively attach to certain compounds. The quantity
or concentration of the compound can then be quantified by measuring the fluorescence
intensity of the label, which is attached to the compound. Such methods are often used to
quantify low concentrations of DNA or RNA, for example.
You can use the Protocol Manager in the SoftMax Pro Software to quickly find and open a
predefined protocol.
More protocols and updated protocols can be downloaded from the Knowledge Base on the
Molecular Devices support web site (www.moleculardevices.com/support) or from the
protocol sharing web site (www.softmaxpro.org).
The following detection cartridges have fluorescence intensity read mode capability:
Tunable Wavelength (TUNE) Detection Cartridge, see page 97
Multi-Mode (MULTI) Detection Cartridge, see page 102
Fluorescence Intensity (FI) Detection Cartridges, see page 116
Fluorescence Intensity (FI) GeneBLAzer Detection Cartridge, see page 119
Fluorescence Intensity Dual Label (FI-DL) (MultiTox-Fluor) Detection Cartridge, see page
122
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Analyzing
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