Three-dimensional imaging with reflection synthetic confocal microscopy.

Biomedical imaging lacks label-free microscopy techniques able to reconstruct the contour of biological cells in solution, in 3D and with high resolution, as required for the fast diagnosis of numerous diseases. Inspired by computational optical coherence tomography techniques, we present a tomographic diffractive microscope in reflection geometry used as a synthetic confocal microscope, compatible with this goal and validated with the 3D reconstruction of a human effector T lymphocyte.

Numerical approach for reducing out-of-focus light in bright-field fluorescence microscopy and superresolution speckle microscopy.

The standard two-dimensional (2D) image recorded in bright-field fluorescence microscopy is rigorously modeled by a convolution process involving a three-dimensional (3D) sample and a 3D point spread function. We show on synthetic and experimental data that deconvolving the 2D image using the appropriate 3D point spread function reduces the contribution of the out-of-focus fluorescence, resulting in a better image contrast and resolution. This approach is particularly interesting for superresolution speckle microscopy, in which the resolution gain stems directly from the efficiency of the deconvolution of each speckle image.

Versatile inversion tool for phaseless optical diffraction tomography.

Estimating three-dimensional complex permittivity of a sample from the intensity recorded at the image plane of a microscope for various angles of illumination, as in optical Fourier ptychography microscopy, permits one to avoid the interferometric measurements of classical tomographic diffraction microscopes (TDMs). In this work, we present a general inversion scheme for processing intensities that can be applied to any microscope configuration (transmission or reflection, low or high numerical aperture), scattering regime (single or multiple scattering), or sample-holder geometries (with or without substrate). The inversion procedure is tested on a wide variety of synthetic experiments, and the reconstructions are compared to that of TDMs. In most cases, phaseless data yield the same result as complex data, thus paving the way toward a drastic simplification of TDM implementation.

Coherent anti-Stokes Raman Fourier ptychography.

We present a theoretical and numerical study of coherent anti-Stokes Raman scattering Fourier ptychography microscopy (CARS-FPM), a scheme that has not been considered so far in the previously reported CARS wide-field imaging schemes. In this approach, the distribution of the Raman scatterer density of the sample is reconstructed numerically from CARS images obtained under various angles of incidences of the pump or Stokes beam. Our inversion procedure is based on an accurate vectorial model linking the CARS image to the sample and yields both the real and imaginary parts of the susceptibility, the latter giving access to the Raman information, with an improved resolution.

Versatile inversion tool for phaseless optical diffraction tomography: publisher's note.

This publisher's note corrects a typo in the title of J. Opt. Soc. Am. A36, C1 (2019)JOAOD60740-323210.1364/JOSAA.36.0000C1.

Multi-wavelength multi-angle reflection tomography.

We have developed a reflection tomographic microscope in which the sample is reconstructed from different holograms recorded under various angles and wavelengths of incidence. We present an iterative inversion algorithm based on a rigorous modeling of the wave-sample interaction that processes all the data simultaneously to estimate the sample permittivity distribution. We show that using several wavelengths permits a significant improvement of the reconstruction, especially along the optical axis.

Two-photon speckle illumination for super-resolution microscopy.

We present a numerical study of a microscopy setup in which the sample is illuminated with uncontrolled speckle patterns and the two-photon excitation fluorescence is collected on a camera. We show that, using a simple deconvolution algorithm for processing the speckle low-resolution images, this wide-field imaging technique exhibits resolution significantly better than that of two-photon excitation scanning microscopy or one-photon excitation bright-field microscopy.

Phase imaging and synthetic aperture super-resolution via total internal reflection microscopy.

Total internal reflection microscopy is mainly used in its fluorescence mode and is the reference technique to image fluorescent proteins in the vicinity of cell membranes. Here, we show that this technique can easily become a phase microscope by simply detecting the coherent signal resulting from the interference between the field scattered by the probed sample and the total internal reflection. Moreover, combining several illumination angles permits generating synthetic aperture reconstructions with improved resolutions compared to standard label-free microscopy techniques.

Improving the axial and lateral resolution of three-dimensional fluorescence microscopy using random speckle illuminations.

We consider a fluorescence microscope in which several three-dimensional images of a sample are recorded for different speckle illuminations. We show, on synthetic data, that by summing the positive deconvolution of each speckle image, one obtains a sample reconstruction with axial and transverse resolutions that compare favorably to that of an ideal confocal microscope.

Superresolution with full-polarized tomographic diffractive microscopy.

Tomographic diffractive microscopy is a three-dimensional imaging technique that reconstructs the permittivity map of the probed sample from its scattered field, measured both in phase and in amplitude. Here, we detail how polarization-resolved measurements permit us to significantly improve the accuracy and the resolution of the reconstructions, compared to the conventional scalar treatments used so far. An isotropic transverse resolution of about 100 nm at a wavelength of 475 nm is demonstrated using this approach.

High-resolution tomographic diffractive microscopy in reflection configuration.

Tomographic diffractive microscopy (TDM) is a label-free imaging technique that reconstructs the 3D refractive index map of the probed object with an improved resolution compared to confocal microscopy. In this work, we consider a TDM implementation in which the sample is deposited on a reflective substrate. We show that this configuration requires calibration and inversion procedures that account for the presence of the substrate for getting highly resolved quantitative reconstructions.

Discrete dipole approximation in time domain through the Laplace transform.

We present a form of the discrete dipole approximation for electromagnetic scattering computations in time domain. We show that the introduction of complex frequencies, through the Laplace transform, significantly improves the computation time. We also show that the Laplace transform and its inverse can be combined to extract the field inside a scatterer at a real resonance frequency.

Discrete dipole approximation for time-domain computation of optical forces on magnetodielectric scatterers.

We present a general approach, based on the discrete dipole approximation (DDA), for the computation of the exchange of momentum between light and a magnetodielectric, three-dimensional object with arbitrary geometry and linear permittivity and permeability tensors in time domain. The method can handle objects with an arbitrary shape, including objects with dispersive dielectric and/or magnetic material responses.

High-resolution total-internal-reflection fluorescence microscopy using periodically nanostructured glass slides.

We compare the performance of a total-internal-reflection fluorescence microscope under varying illumination and substrate conditions. The samples are deposited on a standard homogeneous glass slide or on a grating and illuminated by one or two interfering beams at various incident angles. A conjugate gradient with positivity a priori information is used to reconstruct the fluorophore density from the images. Numerical studies demonstrate that when the sample lies on an optimized grating, the lateral resolution of the microscope is greatly improved, up to fourfold, the best result being obtained when the grating is illuminated by two interfering beams.

Experimental demonstration of quantitative imaging beyond Abbe's limit with optical diffraction tomography.

Optical diffraction tomography (ODT) is a recent imaging technique that combines the experimental methods of phase microscopy and synthetic aperture with the mathematical tools of inverse scattering theory. We show experimentally that this approach permits us to obtain the map of permittivity of highly scattering samples with axial and transverse resolutions that are much better than that of a microscope with the same numerical aperture.

Coupled-dipole method in time domain.

We present a time-domain formulation of electrodynamics based on the self-consistent derivation of the electromagnetic field in a linear, dispersive, lossy object via the coupled dipole method.

Subdiffraction resolution in total internal reflection fluorescence microscopy with a grating substrate.

We propose a fluorescence surface imaging system that presents a power of resolution beyond that of the diffraction limit without resorting to saturation effects or probe scanning. This is achieved by depositing the sample on an optimized periodically nanostructured substrate in a standard total internal reflection fluorescence microscope. The grating generates a high-spatial-frequency light grid that can be moved throughout the sample by changing the incident angle. An appropriate reconstruction procedure permits one to recover the fluorescence amplitude from the images obtained for various incidences. Simulations of this imaging system show that the resolution is not limited by diffraction but by the period of the grating.

Phaseless imaging with experimental data: facts and challenges.

Two-dimensional target characterization using inverse profiling approaches with total-field phaseless data is discussed. Two different inversion schemes are compared. In the first one, the intensity-only data are exploited in a minimization scheme, thanks to a proper definition of the cost functional. Specific normalization and starting guess are introduced to avoid the need for global optimization methods. In the second scheme [J. Opt. Soc. Am. A21, 622 (2004)], one exploits the field properties and the theoretical results on the inversion of quadratic operators to derive a two-step solution strategy, wherein the (complex) scattered fields embedded in the available data are retrieved first and then a traditional inverse scattering problem is solved. In both cases, the analytical properties of the fields allow one to properly fix the measurement setup and identify the more convenient strategy to adopt. Also, indications on the number and types of sources and receivers to be used are given. Results from experimental data show the efficiency of these approaches and the tools introduced.

Two-dimensional inverse profiling problem using phaseless data.

We discuss the characterization of two-dimensional targets based on their diffracted intensity. The target characterization is performed by minimizing an adequate cost functional, combined with a level-set representation if the target is homogeneous. One key issue in this minimization is the choice of an updating direction, which involves the gradient of the cost functional. This gradient can be evaluated using a fictitious field, the solution of an adjoint problem in which receivers act as sources with a specific amplitude. We explore the Born approximation for the adjoint field and compare various approaches for a wide variety of objects.

Influence of multiple scattering on three-dimensional imaging with optical diffraction tomography.

Optical diffraction tomography is an imaging technique that permits retrieval of the map of permittivity of an object from its scattered far field. Most reconstruction procedures assume that single scattering is dominant so that the scattered far field is linearly linked to the permittivity. In this work, we present a nonlinear inversion method and apply it to complex three-dimensional samples. We show that multiple scattering permits one to obtain a power of resolution beyond the classical limit imposed by the use of propagative incident and diffracted waves. Moreover, we stress that our imaging method is robust with respect to correlated and uncorrelated noise.