Fluorometry of turbid and absorbant samples and the membrane fluidity of intact erythrocytes.
01 January 1985
In employing intrinsic or extrinsic fluorophores in the study of whole cells, or other strongly absorbant and/or scattering samples, the measured fluorescence intensity and polarization is seriously affected by absorption and scattering within the sample cuvet. These artifacts are analyzed and simple protocols are provided for overcoming them. The validity of the method is confirmed by experiments in which the emission anisotropies and fluorescence yields of membrane probes in intact erythrocytes was measured with precision. It is also shown that the rotational mobility of 1-phenyl-3- (2-naphthyl)-2-pyrazoline is the same for intact erythrocytes and ghosts.