Elimination of imaging artifacts in second harmonic generation microscopy using interferometry.

Conventional second harmonic generation (SHG) microscopy might not clearly reveal the structure of complex samples if the interference between all scatterers in the focal volume results in artefactual patterns. We report here the use of interferometric second harmonic generation (I-SHG) microscopy to efficiently remove these artifacts from SHG images. Interfaces between two regions of opposite polarity are considered because they are known to produce imaging artifacts in muscle for instance.

Effect of refractive index mismatch on forward-to-backward ratios in SHG imaging.

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.

Gouy phase shift measurement using interferometric second-harmonic generation.

We report on a simple way to directly measure the Gouy phase shift of a strongly focused laser beam. This is accomplished by using a recent technique, namely, interferometric second-harmonic generation. We expect that this method will be of interest in a wide range of research fields, from high-harmonic and attosecond pulse generation to femtochemistry and nonlinear microscopy.

Effects of refractive index mismatch on SRS and CARS microscopy.

An inhomogeneous linear refractive index profile, such as that occurring in biological tissues, is shown to significantly alter stimulated Raman scattering (SRS) and coherent anti-Stokes Raman scattering (CARS) microscopy images. Our finite-difference time-domain simulations show that near-field enhancement and microlensing can lead to an increase of an object's perceived molecular density by a factor of nine and changes in its perceived position by 0.4 μm up to 1.

The Impact of Collagen Fibril Polarity on Second Harmonic Generation Microscopy.

In this work, we report the implementation of interferometric second harmonic generation (SHG) microscopy with femtosecond pulses. As a proof of concept, we imaged the phase distribution of SHG signal from the complex collagen architecture of juvenile equine growth cartilage. The results are analyzed in respect to numerical simulations to extract the relative orientation of collagen fibrils within the tissue.

Analysis of forward and backward Second Harmonic Generation images to probe the nanoscale structure of collagen within bone and cartilage.

Collagen ultrastructure plays a central role in the function of a wide range of connective tissues. Studying collagen structure at the microscopic scale is therefore of considerable interest to understand the mechanisms of tissue pathologies. Here, we use second harmonic generation microscopy to characterize collagen structure within bone and articular cartilage in human knees.