Elimination of polarization bias in fluorescence intensity measurements

Abstract

It is proposed theoretically and confirmed experimentally that, if the fluorescence emission from a polarized sample is viewed in the horizontal plane at an angle of 45° (or 135°) to the direction of propagation of the exciting radiation with a polarizer set at 54.75° (or 125.25°) from the vertical direction, then, the reading obtained is proportional to the total flux emitted by the sample and is independent of the state of polarization of the exciting radiation and the emission anisotropy of the sample. The polarization sensitivity of the emission detection system then becomes a factor which is included in the spectral sensitivity curve of the emission detection system.