High-Speed Tapping Mode AFM Utilizing Recovery of Tip-Sample Interaction

We propose to use the State Estimation by Sum-of-Norms Regularisation (STATESON-)algorithm for recovering the tip-sample interaction in high-speed tapping mode atomic force microscopy (AFM). This approach enables accurate sample height estimation for each independent cantilever oscillation period, provided that the tip-sample interaction...

Adaptive optics in single objective inclined light sheet microscopy enables three-dimensional localization microscopy in adult Drosophila brains

Single-molecule localization microscopy (SMLM) enables the high-resolution visualization of organelle structures and the precise localization of individual proteins. However, the expected resolution is not achieved in tissue as the imaging conditions deteriorate. Sample-induced aberrations distort the point spread function (PSF), and high...

Particle detection using closed-loop active model diagnosis

We demonstrate a novel closed-loop input design technique on the detection of particles in an imaging system such as a fluorescence microscope. The probability of misdiagnosis is minimized while constraining the input energy such that for instance phototoxicity is reduced. The key novelty of the closed-loop design is that each next input is...

Adaptive optics plug-and-play setup for high-resolution microscopes with multi-actuator adaptive lens

Adaptive Optics (AO) has revealed as a very promising technique for high-resolution microscopy, where the presence of optical aberrations can easily compromise the image quality. Typical AO systems however, are almost impossible to implement on commercial microscopes. We propose a simple approach by using a Multi-actuator Adaptive Lens (MAL)...

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...

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...

Adaptive optics for confocal laser scanning microscopy with adjustable pinhole

The pinhole plays an important role in confocal laser scanning microscopy (CLSM) for adaptive optics (AO) as well as in imaging, where the size of the pinhole denotes a trade-off between out-of-focus rejection and wavefront distortion. This contribution proposes an AO system for a commercial CLSM with an adjustable square pinhole to cope with...

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...

Lensless coherent imaging by sampling of the optical field with digital micromirror device

We have experimentally demonstrated a lensless coherent microscope based on direct registration of the complex optical field by sampling the pupil with a sequence of two-point interferometers formed by a digital micromirror device. Complete registration of the complex amplitude in the pupil of the imaging system, without any reference beam,...

Optimization-based wavefront sensorless adaptive optics for multiphoton microscopy

Optical aberrations have detrimental effects in multiphoton microscopy. These effects can be curtailed by implementing model-based wavefront sensorless adaptive optics, which only requires the addition of a wavefront shaping device, such as a deformable mirror (DM) to an existing microscope. The aberration correction is achieved by maximizing a...

Snapshot coherence-gated direct wavefront sensing for multi-photon microscopy

Deep imaging in turbid media such as biological tissue is challenging due to scattering and optical aberrations. Adaptive optics has the potential to compensate the tissue aberrations. We present a wavefront sensing scheme for multi-photon scanning microscopes using the pulsed, near-infrared light reflected back from the sample utilising...