Analysis of the hybrid light field reconstruction and comparison with Richardson-Lucy Light Field Deconvolution.

Conventional microscopes have a high spatial resolution and a low depth-of-field. Light field microscopes have a high depth-of-field but low spatial resolution. A new hybrid approach uses information from both systems to reconstruct a high-resolution light field [Appl. Opt.58, A142 (2019)APOPAI0003-693510.1364/AO.58.00A142]. The resolution of the resulting light field is said to be limited only by diffraction and the size of the pixels. In this paper, we evaluate this method. Using simulation data we compare the output of the hybrid reconstruction algorithm with its simulated ground truth. Our analyses reveal that the observed improvement in the light field quality is not a consequence of data fusion or incorporation of information from a conventional camera, but rather the results of an intermediate interpolation step within the light field itself. This suggests that the required information is already inherent to the light field. By employing the Richardson-Lucy Light Field Deconvolution algorithm, we demonstrate that existing algorithms have already utilized this information.

Higher order wavefront correction and axial scanning in a single fast and compact piezo-driven adaptive lens.

We present a compact adaptive glass membrane lens for higher order wavefront correction and axial scanning, driven by integrated segmented piezoelectric actuators. The membrane can be deformed in a combination of rotational symmetry providing focus control of up to ± 6 m-1 and spherical aberration correction of up to 5 wavelengths and different discrete symmetries to correct higher order aberrations such as astigmatism, coma and trefoil by up to 10 wavelengths. Our design provides a large clear aperture of 12 mm at an outer diameter of the actuator of 18 mm, a thickness of 2 mm and a response time of less than 2 ms.

Fully refractive telecentric f-theta microscope based on adaptive elements for 3D raster scanning of biological tissues.

Various techniques in microscopy are based on point-wise acquisition, which provides advantages in acquiring sectioned images, for example in confocal or two-photon microscopy. The advantages come along with the need to perform three-dimensional scanning, which is often realized by mechanical movement achieved by stage-scanning or piezo-based scanning in the axial direction. Lateral scanning often employs galvo-mirrors, leading to a reflective setup and hence to a folded beam path. In this paper, we introduce a fully refractive microscope capable of three-dimensional scanning, which employs the combination of an adaptive lens, an adaptive prism, and a tailored telecentric f-theta objective. Our results show that this microscope is capable to perform flexible three-dimensional scanning, with low scan-induced aberrations, at a uniform resolution over a large tuning range of X=Y=6300 µ m and Z=480 µ m with only transmissive components. We demonstrate the capabilities at the example of volumetric measurements on the transgenic fluorescence of the thyroid of a zebrafish embryo and mixed pollen grains. This is the first step towards flexible aberration-free volumetric smart microscopy of three-dimensional samples like embryos and organoids, which could be exploited for the demands in both lateral and axial dimensions in biomedical samples without compromising image quality.

Reliability of tunable lenses: feedback sensors and the influence of temperature, orientation, and vibrations.

We compare different aspects of the robustness to environmental conditions of two different types of piezo-actuated fluid-membrane lenses: a silicone membrane lens, where the piezo actuator indirectly deforms the flexible membrane through fluid displacement, and a glass membrane lens, where the piezo actuator directly deforms the stiff membrane. While both lenses operated reliably over the temperature range of 0°-75°C, there was a significant effect on their actuation characteristics, which can be well described through a simple model. The silicone lens in particular showed a variation in focal power of up to 0.1m-1 ∘C-1. We demonstrated that integrated pressure and temperature sensors can provide feedback for focal power, however, limited by the response time of the elastomers in the lenses, with polyurethane in the support structures of the glass membrane lens being more critical than the silicone. Studying the mechanical effects, the silicone membrane lens showed a gravity-induced coma and tilt, and a reduced imaging quality with the Strehl ratio decreasing from 0.89 to 0.31 at a vibration frequency of 100 Hz and an acceleration of 3g. The glass membrane lens was unaffected by gravity, and the Strehl ratio decreased from 0.92 to 0.73 at a vibration of 100 Hz, 3g. Overall, the stiffer glass membrane lens is more robust against environmental influences.

In-line refractive index measurement: a simple method based on image detection.

We present a simple method to determine the refractive index of fluids that is suitable for real-time integrated measurements by imaging a collimated beam through a fluid volume and determining its diameter on a CMOS sensor. Our experimental results agree with the prediction of our analytical model, and the resulting refractive index agrees with the measurements obtained with a commercial refractometer with an RMS deviation of just ±0.003. This method requires only inexpensive components: a light source, two lenses, and a camera sensor; it is suitable for real-time monitoring, and it is essentially unlimited in the range of refractive indices.

Self-Sensing of a Magnetically Actuated Prism.

We demonstrate a method for self-sensing of a magnetically actuated prism that can be used, e.g., in a feedback-loop without the need of additional sensors. In order to use the impedance of the actuation coils as a measurement parameter, we first obtained the optimal measurement frequency that is well separated from the actuation frequencies and at the same time provides the best compromise between sensitivity to the position and robustness. We then developed a combined actuation and measurement driver, and correlated its output signal to the mechanical state of the prism using a defined calibration sequence. We demonstrate that we can reliably measure the state of each actuator and determine the tilt angle of the prism with an accuracy of ±0.1∘ in the polar angle over a range of ±4∘ and ±20 mrad in the azimuthal angle.

Tunable doublets: piezoelectric glass membrane lenses with an achromatic and spherical aberration control.

We present two versions of tunable achromatic doublets based on each two piezoelectrically actuated glass membranes that create the surface of fluid volumes with different dispersions: a straightforward back-to-back and a more intricate stack of the fluid volumes. In both cases, we can control the chromatic focal shift and focal power independently by a suitable combination of actuation voltages on both active membranes. The doublets have a large aperture of 12 mm at an outer diameter of the actuator of 18 mm, an overall thickness of 3 mm and a short response time of around 0.5 ms and, in addition, provide spherical aberration correction. The two designs have an achromatic focal power range of ±2.2 m-1 and ±3.2 m-1 or, for the purpose of actively correcting chromatic errors, a chromatic focal shift at vanishing combined focal power of up to ±0.08 m-1 and ±0.12 m-1.

A quick and accurate method to determine the Poisson's ratio and the coefficient of thermal expansion of PDMS.

We present a new and accurate method to determine the Poisson's ratio of PDMS, using thermal expansion and an optical surface profilometer. The Poisson's ratio of Sylgard 184 was found to be ν = 0.4950 ± 0.0010 and for Sylgard 182, ν = 0.4974 ± 0.0006. Furthermore, we found that for both PDMS types, the coefficient of thermal expansion depends approximately linearly on the curing temperature. This method can be used for almost any kind of soft polymer that can be cured from a liquid at elevated temperatures.

Aspherical high-speed varifocal mirror for miniature catadioptric objectives.

We present a varifocal mirror based on a piezo-actuated glass membrane that can be used as a secondary mirror in miniature Cassegrain-type mirror- or catadioptric objectives. The mirror section has a diameter of 10 mm on a clear membrane diameter of 23 mm, with a focal range of ±8 m-1 and a response time on the millisecond-scale. The two piezo layers enable an aspherical tuning range that covers the elliptic, parabolic and hyperbolic regime over most of the focal range. We demonstrate the application of the mirror in a simple catadioptric telefocus objective with a focal length of 68 mm at an aperture of 22 mm and a thickness of 16.6 mm.

Segmented Bessel beams.

We investigate segmented Bessel beams that are created by placing different ring apertures behind an axicon that is illuminated with a plane wave. We find an analytical estimate to determine the shortest possible beam segment by deriving a scale-invariant analytical model using appropriate dimensionless parameters such as the wavelength and the axicon angle. This is verified using simulations and measurements, which are in good agreement. The size of the ring apertures was varied from small aperture sizes in the Frauhofer diffraction limit to larger aperture sizes in the classical limit.

Phase gratings with tunable diffraction efficiency.

We present a new principle for tuning the diffraction efficiency of an optical grating and its implementation in a micro-optical device. The overlap of two phase gratings is used to vary the effective phase shift and hence the diffraction efficiency. We study the working principle using Fourier Optics to simulate the diffraction pattern in the far field and design and realize a device based on integrated piezo actuation. We find good agreement between simulation and experiment and observe a suppression of the first diffraction order intensity by more than 97% and response times of less than 3 ms.

Volumetric HiLo microscopy employing an electrically tunable lens.

Electrically tunable lenses exhibit strong potential for fast motion-free axial scanning in a variety of microscopes. However, they also lead to a degradation of the achievable resolution because of aberrations and misalignment between illumination and detection optics that are induced by the scan itself. Additionally, the typically nonlinear relation between actuation voltage and axial displacement leads to over- or under-sampled frame acquisition in most microscopic techniques because of their static depth-of-field. To overcome these limitations, we present an Adaptive-Lens-High-and-Low-frequency (AL-HiLo) microscope that enables volumetric measurements employing an electrically tunable lens. By using speckle-patterned illumination, we ensure stability against aberrations of the electrically tunable lens. Its depth-of-field can be adjusted a-posteriori and hence enables to create flexible scans, which compensates for irregular axial measurement positions. The adaptive HiLo microscope provides an axial scanning range of 1 mm with an axial resolution of about 4 µm and sub-micron lateral resolution over the full scanning range. Proof of concept measurements at home-built specimens as well as zebrafish embryos with reporter gene-driven fluorescence in the thyroid gland are shown.

Quasi-Bessel beams from asymmetric and astigmatic illumination sources.

We study the spatial intensity distribution and the self-reconstruction of quasi-Bessel beams produced from refractive axicon lenses with edge emitting laser diodes as asymmetric and astigmatic illumination sources. Comparing these to a symmetric mono-mode fiber source, we find that the asymmetry results in a transition of a quasi-Bessel beam into a bow-tie shaped pattern and eventually to a line shaped profile at a larger distance along the optical axis. Furthermore, we analytically estimate and discuss the effects of astigmatism, substrate modes and non-perfect axicons. We find a good agreement between experiment, simulation and analytic considerations. Results include the derivation of a maximal axicon angle related to astigmatism of the illuminating beam, impact of laser diode beam profile imperfections like substrate modes and a longitudinal oscillation of the core intensity and radius caused by a rounded axicon tip.

Bio-inspired variable imaging system simplified to the essentials: modelling accommodation and gaze movement.

A combination of an aspherical hybrid diffractive-refractive lens with a flexible fluidic membrane lens allows the implementation of a light sensitive and wide-aperture optical system with variable focus. This approach is comparable to the vertebrate eye in air, in which the cornea offers a strong optical power and the flexible crystalline lens is used for accommodation. Also following the natural model of the human eye, the decay of image quality with increasing field position is compensated, in the optical system presented here, by successively addressing different tilting angles which mimics saccadic eye-movements. The optical design and the instrumental implementation are presented and discussed, and the working principle is demonstrated.

Axial scanning in confocal microscopy employing adaptive lenses (CAL).

In this paper we analyze the capability of adaptive lenses to replace mechanical axial scanning in confocal microscopy. The adaptive approach promises to achieve high scan rates in a rather simple implementation. This may open up new applications in biomedical imaging or surface analysis in micro- and nanoelectronics, where currently the axial scan rates and the flexibility at the scan process are the limiting factors. The results show that fast and adaptive axial scanning is possible using electrically tunable lenses but the performance degrades during the scan. This is due to defocus and spherical aberrations introduced to the system by tuning of the adaptive lens. These detune the observation plane away from the best focus which strongly deteriorates the axial resolution by a factor of ~2.4. Introducing balancing aberrations allows addressing these influences. The presented approach is based on the employment of a second adaptive lens, located in the detection path. It enables shifting the observation plane back to the best focus position and thus creating axial scans with homogeneous axial resolution. We present simulated and experimental proof-of-principle results.

Fast and robust piezoelectric axicon mirror.

In this Letter, we demonstrate the first high-speed piezoelectric axicon mirror. We achieve a usable aperture of 10 mm up to the maximum radius of the robust, 300 µm thick mirror substrate using a floating boundary condition. The highly aspheric, conical shape is programmed into the device by ring-shaped electrodes, for which we have developed an automated optimization strategy for their individual electrode potentials. In addition, we developed a simple control circuit, in which the conical profile can be programmed and adjusted with just one control signal. The device is fabricated by rapid prototyping to avoid cleanroom processing. The tunable mirror features a resonance frequency of 10 kHz and a static deflection of 5.8 µm at a surface deviation of 63 nm, and is thus able to generate a quasi-Bessel beam.

Tunable MEMS axicon mirror arrays.

Highly aspheric reflective micro-optics for the generation of quasi-nondiffracting beams are of great interest for a wide variety of applications. However, up to now it was impossible to fabricate tunable arrays of these elements. In this Letter, we demonstrate the first array of purely reflective tunable microelectromechanical systems (MEMS) microaxicon mirrors with a conical shape and a continuous surface. The actuation is achieved by thermal expansion in a solid state design and the tuning range allows for large conical angles and is able to form concave as well as convex axicons. The deflection of the mirror surface and the propagation of the resulting quasi-Bessel beams have been characterized to prove the functionality of the device.

Sub-3-cycle vortex pulses of tunable topological charge.

Novel types of reflective spiral micro-electro-mechanical systems were used to generate few-cycle vortex pulses of variable topological charge from a Ti:sapphire laser oscillator. The phase profile of these components was controlled by varying the temperature. The temporal properties of the pulses were characterized with spatially resolved nonlinear autocorrelation. The beam structure resembles a slightly distorted Laguerre-Gaussian distribution. The different topological charges were indicated by detecting Poynting-vector maps with a programmable Shack-Hartmann sensor of enhanced angular sensitivity.

A Miniaturized Piezo Stack Impact Actuation Mechanism for Out-of-Plane Freely Moveable Masses.

We present the prototype and analytical model of a miniaturized impact actuation mechanism, providing a fast out-of-plane displacement to accelerate objects against gravity, allowing for freely moving objects and hence for large displacements without the need for cantilevers. To achieve the necessary high speed, we chose a piezoelectric stack actuator driven by a high-current pulse generator, connected to a rigid support and a rigid three-point contact with the object. We describe this mechanism with a spring-mass model and compare various spheres with different masses and diameters and from different materials. As expected, we found that larger flight heights are achieved by harder spheres, achieving, e.g., approx. 3 mm displacement for a 3 mm steel sphere using a 3 × 3 × 2 mm3 piezo stack.

Antireflective "moth-eye" structures on tunable optical silicone membranes.

Flexible silicone membranes are key components for tunable optical lenses. The elastic operation of the membranes impedes the use of classical layer systems for an antireflective (AR) effect. To overcome this limitation, we equipped optical elastomer membranes with "moth-eye" structures directly in the flexible silicone substrate. The manufacturing of the AR structures in the flexible membrane includes a mastering process based on block copolymer micelle nanolithography followed by a replication method. We investigate the performance of the resulting AR structures under strain of up to 20% membrane expansion. A significant transmittance enhancement of up to 2.5% is achieved over the entire visible spectrum, which means that more than half of the surface reflection losses are compensated by the AR structures.