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Holographic microscopy has developed into a powerful tool for 3D particletracking, yielding nanometer-scale precision at high frame rates. However,current particle tracking algorithms ignore the effect of the microscopeobjective on the formation of the recorded hologram. As a result, particletracking in holographic microscopy is currently limited to particles well abovethe microscope focus. Here, we show that modeling the effect of anaberration-free lens allows tracking of particles above, near, and below thefocal plane in holographic microscopy, doubling the depth of field. Finally, weuse our model to determine the conditions under which ignoring the effect ofthe lens is justified, and in what conditions it leads to systematic errors.

Large depth-of-field tracking of colloidal spheres in holographic microscopy by modeling the objective lens