Three-dimensional imaging through patterned type-1 microscopy

We report a scanning non-confocal fluorescence microscopy scheme that provides images with optical sectioning and with a lateral resolution that surpasses by a factor of two the diffraction resolution limit. This technique is based on the type-1 microscopy concept combined with patterned illumination. The method does not require the application of phase-shifting or post-processing algorithms and provides artifact-free superresolved 3D images. We have validated the theory by means of experimental data

Fast and robust phase-shift estimation in two-dimensional structured illumination microscopy

A method of determining unknown phase-shifts between elementary images in two-dimensional Structured Illumination Microscopy (2D-SIM) is presented. The proposed method is based on the comparison of the peak intensity of spectral components. These components correspond to the inherent structured illumination spectral content and the residual compo- nent that appears from wrongly estimated phase-shifts. The estimation of the phase-shifts is carried out by finding the absolute maximum of a function defined as the normalized peak intensity difference in the Fourier domain. This task is performed by an optimization method providing a fast estimation of the phase-shift. The algorithm stability and robustness are tested for various levels of noise and contrasts of the structured illumination pattern. Furthermore, the proposed approach reduces the number of computations compared to other existing techniques. The method is supported by the theoretical calculations and validated by means of simulated and experimental results

3D Integral Microscopy based in far-field detection

Lately, Integral-Imaging systems have shown very promising capabilities of capturing the 3D structure of microscopic and macroscopic scenes. The aim of this work is to provide an optimal design for 3D-integral microscopy with extended depth of eld and enhanced lateral resolution. By placing an array of microlenses at the aperture stop of the objective, this setup provides a set of orthographic views of the 3D sample. Adopting well known integral imaging reconstruction algorithms it can be shown that the depth of eld as well as spatial resolution are improved with respect to conventional integral microscopy imaging. Our claims are supported on theoretical basis and experimental images of a resolution test target, and biological samples

Integral imaging with Fourier-plane recording

Integral Imaging is well known for its capability of recording both the spatial and the angular information of threedimensional (3D) scenes. Based on such an idea, the plenoptic concept has been developed in the past two decades, and therefore a new camera has been designed with the capacity of capturing the spatial-angular information with a single sensor and after a single shot. However, the classical plenoptic design presents two drawbacks, one is the oblique recording made by external microlenses. Other is loss of information due to diffraction effects. In this contribution report a change in the paradigm and propose the combination of telecentric architecture and Fourier-plane recording. This new capture geometry permits substantial improvements in resolution, depth of field and computation time

Evaluation of the use of wavefront encoding to reduce depth-induced aberration in structured-illumination microscopy

Three-dimensional imaging is affected by depth-induced spherical aberration (SA) when imaging deep into an optically thick sample. In this work, we evaluate the impact of SA on the performance of incoherent grating-projection structured illumination microscopy (SIM). In particular, we analyze the reduction of the contrast in the structured pattern and compare the reconstructed SIM images for different amounts of SA. In order to mitigate the impact of SA, we implement and evaluate in SIM a wavefront encoded imaging system using a square cubic (SQUBIC) phase mask, an approach shown previously to be successful in conventional microscopy