Matthias Hillenbrand

Thin-sheet laser imaging microscopy for optical sectioning of thick tissues

We report the development of a modular and optimized thin-sheet laser imaging microscope (TSLIM) for nondestructive optical sectioning of organisms and thick tissues such as the mouse cochlea, zebrafish brain/inner ear, and rat brain at a resolution that is comparable to wide-field fluorescence microscopy. TSLIM optically sections tissue using a thin sheet of light by inducing a plane of fluorescence in transparent or fixed and cleared tissues. Moving the specimen through the thinnest portion of the light sheet and stitching these image columns together results in optimal resolution and focus across the width of a large specimen. Dual light sheets and aberration-corrected objectives provide uniform section illumination and reduce absorption artifacts that are common in light-sheet microscopy. Construction details are provided for duplication of a TSLIM device by other investigators in order to encourage further use and development of this important technology.

Tunable hyperchromatic lens system for confocal hyperspectral sensing

A new approach for confocal hyperspectral sensing based on the combination of a diffractive optical element and a tunable membrane fluidic lens is demonstrated. This highly compact lens system is designed to maximize the longitudinal chromatic aberration and select a narrow spectral band by spatial filtering. Changing the curvature of the fluidic lens allows the selected band to be scanned over the whole given spectrum. A hybrid prototype with an integrated electro-magnetic micro-actuator has been realized to demonstrate the functionality of the system. Experimental results show that the spectrum transmitted by the system can be tuned over the entire visible wavelength range, from 450 to 900 nm with a narrow and almost constant linewidth of less than 15 nm. Typical response time for scanning the spectrum by 310 nm is less than 40 ms and the lens system shows a highly linear relationship with the driving current.

Numerical solution of nonparaxial scalar diffraction integrals for focused fields

In this paper, we present sampling conditions for fast-Fourier-transform-based field propagations. The input field and the propagation kernel are analyzed in a combined manner to derive sampling criteria that guarantee accurate calculation results in the output plane. These sampling criteria are also applicable to the propagation of general fields. For focal field calculations, geometrical optics is used to obtain a priori knowledge about the input and output fields. This a priori knowledge is used to determine an optimum balance between computational load and calculation accuracy. In a numerical example, correct results are obtained even though both the input field and the propagation kernel are sampled below the Nyquist rate. Finally, we show how chirp z-transform-based zoom-algorithms may be analyzed using the same techniques.

Fast nonparaxial scalar focal field calculations

An efficient algorithm for calculating nonparaxial scalar field distributions in the focal region of a lens is discussed. The algorithm is based on fast Fourier transform implementations of the first Rayleigh-Sommerfeld diffraction integral and assumes that the input field at the pupil plane has a larger extent than the field in the focal region. A sampling grid is defined over a finite region in the output plane and referred to as a tile. The input field is divided into multiple separate spatial regions of the size of the output tile. Finally, the input tiles are added coherently to form a summed tile, which is propagated to the output plane. Since only a single tile is propagated, there are significant reductions of computational load and memory requirements. This method is combined either with a subpixel sampling technique or with a chirp z-transform to realize smaller sampling intervals in the output plane than in the input plane. For a given example the resulting methods enable a speedup of approximately 800× in comparison to the normal angular spectrum method, while the memory requirements are reduced by more than 99%.

Aberration analysis of optimized Alvarez–Lohmann lenses

In this paper aberrations in Alvarez-Lohmann lenses are analyzed, and a semi-analytical strategy for compensation is derived. An x-y polynomial model is used to describe the aberrations and classify them into static and dynamic components. The lenses are enhanced by higher-order polynomials, and a numerical optimization process is used to determine the most influential coefficients. Two simulations of corrected systems are presented. The first one is optimized for on-axis imaging. The second system is optimized for multiple field points and shows the limitations of a single Alvarez-Lohmann lens. Two systems overcoming these limitations by introducing additional optical surfaces are presented, and their performance is analyzed in simulations.

Chromatic information coding in optical systems for hyperspectral imaging and chromatic confocal sensing

Dispersion causes the focal lengths of refractive and diffractive optical elements to vary with wavelength. In our contribution we show how it can be used for chromatic encoding and decoding of optical signals. We specifically discuss how these concepts can be applied for the implementation of systems with applications in the growing fields of hyperspectral imaging and chromatic distance coding. Refractive systems as well as hybrid combinations of diffractive and refractive elements are used to create specific chromatic aberrations of the sensors. Our design approach enables the tailoring of the sensor properties to the measurement problem and assists designers in finding optimized solutions for industrial applications. The focus of our research is on parallelized imaging systems that cover extended objects. In comparison to point sensors, such systems promise reduced image acquisition times and an increased overall performance. Concepts for three-dimensional profilometry with chromatic confocal sensor systems as well as spectrally resolved imaging of object scenes are discussed.

Hybrid hyperchromats for chromatic confocal sensor systems

Abstract The combination of diffractive and refractive elements in hybrid optical systems allows for precise control of the longitudinal chromatic aberration. We provide comprehensive design strategies for hybrid hyperchromatic lenses that maximise the longitudinal chromatic aberrations. These lenses are mainly used in chromatic confocal sensor systems for efficient non-contact profilometry as well as for measurements of distances and wall thicknesses of transparent materials. Our design approach enables the tailoring of the sensor properties to the specific measurement problem and assists designers in finding optimised solutions for industrial applications. We, for example, demonstrate a hybrid system that significantly exceeds the longitudinal chromatic aberration of purely diffractive elements.

Parallelized chromatic confocal sensor systems

In this paper we present chromatic confocal distance sensors for the parallelized evaluation at several lateral positions. The multi-point measurements are performed using either one- or two-dimensional detector arrays. The first sensor combines the concepts of confocal matrix sensing and snapshot hyperspectral imaging to image a two-dimensional array of laterally separated points with one single shot. In contrast to chromatic confocal matrix sensors which use an RGB detector our system works independently from the spectral reflectivity of the surface under test and requires no object-specific calibration. Our discussion of this sensor principle is supported by experimental results. The second sensor is a multipoint line sensor aimed at high speed applications with frame rates of several thousand frames per second. To reach this evaluation speed a one-dimensional detector is employed. We use spectral multiplexing to transfer the information from different measurement points through a single fiber and evaluate the spectral distribution with a conventional spectrometer. The working principle of the second sensor type is demonstrated for the example of a three-point sensor.

Advanced phase plates for confocal hyperspectral imaging systems

We present innovative confocal hyperspectral imaging systems. The focal length of the systems can be adjusted to change the filtered wavelength band. The active focus variation is achieved by hyperchromatic Alvarez-Lohmann-phase plates

Spectrally multiplexed chromatic confocal multipoint sensing

We present a concept for chromatic confocal distance sensing that employs two levels of spectral multiplexing for the parallelized evaluation of multiple lateral measurement points; at the first level, the chromatic confocal principle is used to encode distance information within the spectral distribution of the sensor signal. For lateral multiplexing, the total spectral bandwidth of the sensor is split into bands. Each band is assigned to a different lateral measurement point by a segmented diffractive element. Based on this concept, we experimentally demonstrate a chromatic confocal three-point sensor that is suitable for harsh production environments, since it works with a single-point spectrometer and does not require scanning functionality. The experimental system has a working distance of more than 50 mm, a measurement range of 9 mm, and an axial resolution of 50 μm.