Fluorescence Polarisation

The fluorescence polarisation option uses a calcite prism polariser, with transparency down to 220 nm, to deliver polarised light to the observation cell. Dichroic sheet polarisers allow only light of a particular plane through to the photomultipliers either side of the cell (parallel and perpendicular to the parallel excitation light). The sensitivities of the two channels are balanced allowing the Kinetic Studio software to accurately calculate and report the fluorescence, polarisation and anisotropy of the cell solution.

When configured for fluorescence polarisation, the sample handling unit is configured with two photomultipliers mounted on the cell block in a T format. Two emission polarizers are used, one for parallel and one for perpendicular emission. The software automatically processes the data to calculate anisotropy and polarisation making the procedure straightforward and intuitive. Single or multiple shots can be acquired using the Kinetic Studio ‘series’ feature to automate and if required average a number of shots.

Relevant Application Notes

• AN.012.S60

  Binding studies of a DNA repair enzyme using the SFA-20 Stopped-Flow fluorescence anisotropy

  Keywords

  1. Fluorescence polarisation
  2. stopped-flow
  3. anisotropy
  4. DNA-protein interactions