Multifocal microscopes (MFMs) are becoming increasingly popular in fluorescence microscopy due to their high speed three-dimensional (3D) imaging capabilities. Conventional MFMs use a fixed fabricated grating as the multifocal grating but these are limited to a restricted wavelength range and a fixed object-plane separation. Spatial light modulators (SLMs) represent an alternative to fabricated gratings due to their real-time programmability, providing complete control over emission wavelength range and object plane separations. However, algorithms commonly used to obtain multifocal grating patterns which provide uniform intensity across the subimages are not directly applicable to SLM-based MFMs due to inherent pixel-to-pixel crosstalk effects present in the SLM chip. We recently developed an in-situ iterative algorithm which generates grating patterns that provide near-uniform illumination of the subimages in SLM-based MFMs. This algorithm is universal across wavelengths, object-plane separations, and SLM manufacturers. As part of our efforts to develop an SLM-based MFM that can respond rapidly to changing experimental parameters, we implement a gradient descent-based optimization method. We evaluate its performance in comparison with a grid search based routine. Experimental results obtained on a custom-made SLM-based MFM indicate that the grid-search optimized grating patterns provide superior subimage intensity uniformity versus the gradient-descent method. These experiments also provide an insight into the energy landscape involved in these optimizations. This study increases the utility of SLM-based MFMs in high-speed imaging.
Biological processes such as processive enzyme turnover and intracellular cargo trafficking involve the dynamic motion of a small ”article" along a curvilinear biopolymer track. To understand these processes that occur across multiple length and time scales, one must acquire both the trajectory of the particle and the position of the track along which it moves, possibly by combining high-resolution single-particle tracking with conventional microscopy. Yet, usually there is a significant resolution mismatch between these modalities: while the tracked particle is localized with a precision of 10 nm, the image of the surroundings is limited by optical diffraction, with 200 nm lateral and 500 nm axial resolutions. Compared to the particle's trajectory, the surrounding curvilinear structure appears as a blurred and noisy image. This disparity in the spatial resolutions of the particle trajectory and the surrounding curvilinear structure image makes data reconstruction, as well as interpretation, particularly challenging. Analysis is further complicated when the curvilinear structures are oriented arbitrarily in 3D space. Here, we present a prior-apprised unsupervised learning (PAUL) approach to extract information from 3D images where the underlying features resemble a curved line such as a filament or microtubule. This three-stage framework starts with a Hessian-based feature enhancement, which is followed by feature registration, where local line segments are detected on repetitively sampled subimage tiles. In the final stage, statistical learning, segments are clustered based on their geometric relationships. Principal curves are then approximated from each segment group via statistical tools including principal component analysis, bootstrap and kernel transformation. This procedure is characterized on simulated images, where sub-voxel medium deviations from true curves have been achieved. The 3D PAUL approach has also been implemented for successful line localization in experimental 3D images of gold nanowires obtained using a multifocal microscope. This work not only bridges the resolution gap between two microscopy modalities, but also allows us to conduct 3D super line-localization imaging experiments, without using super-resolution techniques.
Since 1997, we have proposed and demonstrated the use of the Texas Instrument (TI) Digital Micromirror Device (DMD) for various non-display applications including optical switching and imaging. In 2009, we proposed the use of the DMD to realize wavefront splitting interferometers as well as a variety of imagers. Specifically, proposed were agile electronically programmable wavefront splitting interferometer designs using a Spatial Light Modulator (SLM) such as (a) a transmissive SLM, (b) a DMD SLM and (c) a Beamsplitter with a DMD SLM. The SLMs operates with on/off or digital state pixels, much like a black and white state optical window to control passage/reflection of incident light. SLM pixel locations can be spatially and temporally modulated to create custom wavefronts for near-common path optical interference at the optical detectors such as a CCD/CMOS sensor, a Focal Plane Array (FPA) sensor or a point-photodetector. This paper describes the proposed DMD-based wavefront splitting interferometer and imager designs and their relevant experimental results.
This paper describes a motionless active Depth from Defocus (DFD) system design suited for long working range camera autofocus applications. The design consists of an active illumination module that projects a scene illuminating coherent conditioned optical radiation pattern which maintains its sharpness over multiple axial distances allowing an increased DFD working distance range. The imager module of the system responsible for the actual DFD operation deploys an electronically controlled variable focus lens (ECVFL) as a smart optic to enable a motionless imager design capable of effective DFD operation. An experimental demonstration is conducted in the laboratory which compares the effectiveness of the coherent conditioned radiation module versus a conventional incoherent active light source, and demonstrates the applicability of the presented motionless DFD imager design. The fast response and no-moving-parts features of the DFD imager design are especially suited for camera scenarios where mechanical motion of lenses to achieve autofocus action is challenging, for example, in the tiny camera housings in smartphones and tablets. Applications for the proposed system include autofocus in modern day digital cameras.
To the best of our knowledge, proposed is a novel variable depth of field smart imager design using intelligent laser targeting for high productivity multiple barcodes reading applications. System smartness comes via the use of an Electronically Controlled Variable Focal-Length Lens (ECVFL) to provide an agile pixel (and/or pixel set) within the laser transmitter and optical imaging receiver. The ECVFL in the receiver gives a flexible depth of field that allows clear image capture over a range of barcode locations. Imaging of a 660 nm wavelength laser line illuminated 95-bit one dimensional barcode is experimentally demonstrated via the smart imager for barcode target distances ranging from 10 cm to 54 cm. The smart system captured barcode images are evaluated using a proposed barcode reading algorithm. Experimental results after computer-based post-processing show a nine-fold increase in barcode target distance variation range (i.e., range variation increased from 2.5 cm to 24.5 cm) when compared to a conventional fixed lens imager. Applications for the smart imager include industrial multiple product tracking, marking, and inspection systems.
Proposed is a smart optical writing head design suitable for high precision industrial laser based machining and manufacturing applications. The design uses an Electronically Controlled Variable Focus Lens (ECVFL) which enables the highest achievable spatial resolution of writing head spot sizes for axial target distances reaching 8 meters. A proof-of-concept experiment is conducted using a visible wavelength laser with a collimated beam that is coupled to beam conditioning optics which includes an electromagnetically actuated deformable membrane liquid ECVFL cascaded with a bias convex lens of fixed focal length. Electronic tuning and control of the ECVFL keeps the laser writing head far-field spot beam radii under 1 mm that is demonstrated over a target range of 20 cm to 800 cm. Applications for the proposed writing head design, which can accommodate both continuous wave and pulsed wave sources, include laser machining, high precision industrial molding of components, as well as materials processing requiring material sensitive optical power density control.
Proposed is a novel eye vision system that combines the use of advanced micro-optic and microelectronic technologies
that includes programmable micro-optic devices, pico-projectors, Radio Frequency (RF) and optical wireless
communication and control links, energy harvesting and storage devices and remote wireless energy transfer capabilities.
This portable light weight system can measure eye refractive powers, optimize light conditions for the eye under test,
conduct color-blindness tests, and implement eye strain relief and eye muscle exercises via time sequenced imaging.
Described is the basic design of the proposed system and its first stage system experimental results for vision spherical
lens refractive error correction.
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