Tomography of fluctuating biological interfaces using defocusing microscopy

Abstract

We show that a bright-field defocused microscope is effectively a phase-contrast microscope, but with advantages over the conventional one and maintaining the same optical resolution. In a multilayered transparent object, the height amplitude (static and dynamic) of each interface can be measured separately with nanometer sensitivity. By scanning the position of the objective focal plane in relation to the surfaces of a red blood cell, we obtain quantitative information on height fluctuations from each surface individually, which can be analyzed with our model of a defocused microscope and compared with theoretical models.