Molecular Devices SpectraMax Paradigm User Manual page 126

SpectraMax Paradigm Multi-Mode Detection Platform User Guide
About Fluorescence Polarization
Fluorescence polarization (FP) mode measures the relative change of polarization of emitted
fluorescent compared to excitation light.
Fluorescence polarization detection is similar to fluorescence intensity, with the important
difference that it uses plane-polarized light, rather than non-polarized light. Plate readers
measure FP of the sample by detecting light emitted both parallel and perpendicular to the
plane of excitation.
By using a fluorescent dye to label a small molecule, its binding to a different molecule of
equal or greater size can be monitored through its speed of rotation.
When molecules are excited with polarized light, the polarization of the emitted light
depends on the size of the molecule to which the fluorophore is bound. Larger molecules
emit a higher percentage of polarized light, while smaller molecules emit a lower percentage
of polarized light because of their rapid molecular movement. For this reason FP is generally
used for molecular binding assays in high-throughput screening (HTS).
For more information on fluorescence polarization, see
Mode on page
Microplate Recommendations
The following information is specific to this detection cartridge. For general microplate
selection and handling guidelines, see
Table 5-20: Microplate Selection Guidelines for the Fluorescence Polarization (FP)
Detection Cartridge
Read Mode
Fluorescence
Polarization (FP)
126
49.
Microplate Other Considerations
Type
Solid black When an application specifies a surface treatment, use only
microplates with the correct treatment.
Molecular Devices recommends using unlidded plate for
fluorescence polarization measurements.
Fluorescence Polarization Read
Selecting Suitable Microplate Types on page 91.
5014038 E