Abstract

Nonlinear optical imaging in the epi-direction is used to image subresolution features. We find that a refractive index mismatch between the object to be imaged and the background medium can change the far-field intensity image. As an example, we study second harmonic generation (SHG) microscopy where the forward-to-backward (F/B) ratio is used to quantify subresolution features. We show both theoretically and experimentally that the inhomogeneous refractive index in collagen tendon tissue creates near-field effects, which can change the F/B ratio by ∼20%-25%, even though the effect is negligible for most of the individual fibrils in the tissue. This is caused by the sensitivity of the backward signal on phase matching conditions.

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