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.
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.
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%.
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.
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.
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