Anisoplanatic adaptive optics in parallelized laser scanning microscopy

Inhomogeneities in the refractive index of a biological microscopy sample can introduce phase aberrations, severely impairing the quality of images. Adaptive optics can be employed to correct for phase aberrations and improve image quality. However, conventional adaptive optics can only correct a single phase aberration for the whole field of...

Physical and Computational Approaches to Aberration Correction In Fluorescence Microscopy

The goal of this thesis, called Physical and Computational Approaches to Aberration Correction In Fluorescence Microscopy, concerns itself with the development of new techniques to control adaptive fluorescence microscopes, so that they can adapt and image with increased resolution, contrast and speed inside complex three-dimensional biological...

Practical guidelines for implementing adaptive optics in fluorescence microscopy

In life sciences, interest in the microscopic imaging of increasingly complex three dimensional samples, such as cell spheroids, zebrafish embryos, and in vivo applications in small animals, is growing quickly. Due to the increasing complexity of samples, more and more life scientists are considering the implementation of adaptive optics in...

Use of digital micromirror devices as dynamic pinhole arrays for adaptive confocal fluorescence microscopy

In this work, we present a new confocal laser scanning microscope capable to perform sensorless wavefront optimization in real time. The device is a parallelized laser scanning microscope in which the excitation light is structured in a lattice of spots by a spatial light modulator, while a deformable mirror provides aberration correction and...

Pupil mask diversity for image correction in microscopy

Three-dimensional microscopy suffers from sample-induced aberrations that reduce the resolution and lead to misinterpretations of the object distribution. In this paper, the resolution of a three-dimensional fluorescent microscope is significantly improved by introducing an amplitude diversity in the form of a binary amplitude mask positioned...

Optimal model-based sensorless adaptive optics for epifluorescence microscopy

We report on a universal sample-independent sensorless adaptive optics method, based on modal optimization of the second moment of the fluorescence emission from a point-like excitation. Our method employs a sample-independent precalibration, performed only once for the particular system, to establish the direct relation between the image...

Hybrid adaptive and computational light-sheet fluorescence microscopy

The light-sheet fluorescence microscopy is an excellent tool for the investigation of large three dimensional microscopy samples at the cellular level, however, the ability to resolve features is strongly affected by the presence of scattering and aberrations. These effects are two fold in light-sheet microscopy, as the illumination path...

Rapid identification of coherent pupil functions from multiple intensity measurements

By taking multiple input-output measurements, it is shown how to determine the input to an optical system that corrects unknown phase aberrations without interferometric measurements or online iterative optimization within a couple of seconds. It is shown to work in simulations and experiment. This technique may also be used to acquire the...

Blind multi-frame deconvolution by tangential iterative projections (TIP)

A methodology for retrieving the unknown object distribution and point-spread functions (PSFs) from a set of images acquired in the presence of temporal phase aberrations is presented in this paper. The method works by finding optimal complimentary linear filters for multi-frame deconvolution. The algorithm uses undemanding computational...

Light-sheet optimization for microscopy

Aberrations, scattering and absorption degrade the performance light-sheet fluorescence microscopes (LSFM). An adaptive optics system to correct for these artefacts and to optimize the light-sheet illumination is presented. This system allows a higher axial resolution to be recovered over the field-of-view of the detection objective. It is...

Adaptive optics in digital micromirror based confocal microscopy

This proceeding reports early results in the development of a new technique for adaptive optics in confocal microscopy. The term adaptive optics refers to the branch of optics in which an active element in the optical system is used to correct inhomogeneities in the media through which light propagates. In its most classical form, mostly used in...