RESUMO
Structured illumination microscopy (SIM) is a powerful technique for obtaining super-resolved fluorescence maps of samples, but it is very sensitive to aberrations or misalignments affecting the excitation patterns. Here, we present a reconstruction algorithm that is able to process SIM data even if the illuminations are strongly distorted. The approach is an extension of the recent blind-SIM technique, which reconstructs simultaneously the sample and the excitation patterns without a priori information on the latter. Our algorithm was checked on synthetic and experimental data using distorted and nondistorted illuminations. The reconstructions were similar to that obtained by up-to-date SIM methods when the illuminations were periodic and remained artifact-free when the illuminations were strongly distorted.
Assuntos
Algoritmos , Artefatos , Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/métodos , Iluminação/métodos , Microscopia de Fluorescência/métodosRESUMO
Tomographic diffractive microscopy is a recent imaging technique that reconstructs quantitatively the three-dimensional permittivity map of a sample with a resolution better than that of conventional wide-field microscopy. Its main drawbacks lie in the complexity of the setup and in the slowness of the image recording as both the amplitude and the phase of the field scattered by the sample need to be measured for hundreds of successive illumination angles. In this Letter, we show that, using a wavefront sensor, tomographic diffractive microscopy can be implemented easily on a conventional microscope. Moreover, the number of illuminations can be dramatically decreased if a constrained reconstruction algorithm is used to recover the sample map of permittivity.
RESUMO
We demonstrate that the axial resolution of a reflection tomographic diffractive microscope is drastically improved when the sample is placed in front of a perfect mirror. We show analytically and with rigorous simulations that this approach permits us to obtain images with the same isotropic resolution as that obtained when the sample is illuminated and observed from every possible angle. The main difficulty lies in accounting properly for the mirror in the inversion algorithm.