SpectraMax Mini User Guide
32
5090339 A
500
550
600
650
0
0.5
1.0
Excitation maximum
of fluorophore
Emission maximum
of fluorophore
R
el
ati
ve
F
lu
or
esce
n
ce
Wavelength (nm)
Excitation
reading wavelength
Emission
reading wavelength
Fluorescence intensity data is dependent on several variables.
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 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.
Luminescence Read Mode
Luminescence is the emission of light by processes that derive energy from essentially non-
thermal changes, the motion of subatomic particles, or the excitation of an atomic system by
radiation. Luminescence detection relies on the production of light from a chemical reaction in a
sample.
In luminescence (LUM) read mode, no excitation is necessary as the measured species emit
light naturally. For this reason, the lamp does not flash, so no background excitation
interference occurs.
For the luminescence read mode, the instrument provides measurements in Relative Light Units
(RLUs).
To help eliminate background luminescence from a plate that has been exposed to light, you
should dark adapt the plate by placing the sample-loaded plate inside the instrument for
several minutes before you start the read.
