A microscope configuration for nanometer 3-D movement monitoring accuracy

In this paper we present a new microscopy configuration based upon temporal tracking of a secondary reflected speckle by imaging the speckle through properly defocused optics. The configuration is used to monitor three-dimensional (3-D) spontaneous contraction of rat cardiac muscle cells while achieving nanometer tracking accuracy at a rate of 30 frames per second (fps) without using interferometric recording. Estimation of the change in the optical path of accuracy of 50 nm in the transverse direction and of 200 nm in the axial direction was achieved.