Holographic Illumination Research Articles

High-axial-resolution speckle-free holographic reconstruction via cylindrical quadratic phase method and temporal focusing.

Holographic techniques enable precise laser manipulation, but suffer from two considerable limitations: speckle and deterioration of axial distribution. Here, we propose a cylindrical quadratic phase (CQP) method with temporal focusing (TF) to generate speckle-free holographic illumination with high axial resolution. TF-CQP utilizes a superposed cylindrical phase as the initial guess to iteratively optimize phase hologram, realizing speckle-free holographic reconstruction on the target focal plane and eliminating secondary focus on the defocused planes. TF-CQP further disperses defocused beams symmetrically by a blazed grating, placed conjugate to the focal plane, which enhances axial confinement. Simulation and experimental results show that TF-CQP reconstructs speckle-free illumination with arbitrary shapes and <10 µm axial resolution. Compared to TF-GS (Gerchberg-Saxton algorithm), widely used in holographic optogenetics, TF-CQP shows increased uniformity of 200% and improved modulation efficiency of 32.33% for parallel holographic illumination, as well as a 10% increment in axial resolution.

Descanned fast light targeting (deFLiT) two-photon optogenetics.

Two-photon light-targeting optogenetics allows controlling selected subsets of neurons with near single-cell resolution and high temporal precision. To push forward this approach, we recently proposed a fast light-targeting strategy (FLiT) to rapidly scan multiple holograms tiled on a spatial light modulator (SLM). This allowed generating sub-ms timely-controlled switch of light patterns enabling to reduce the power budget for multi-target excitation and increase the temporal precision for relative spike tuning in a circuit. Here, we modified the optical design of FLiT by including a de-scan unit (deFLiT) to keep the holographic illumination centered at the middle of the objective pupil independently of the position of the tiled hologram on the SLM. This enables enlarging the number of usable holograms and reaching extended on-axis excitation volumes, and therefore increasing even further the power gain and temporal precision of conventional FLiT.

Holographic Optogenetic Activation of Neurons Eliciting Locomotion in Head-Embedded Larval Zebrafish.

Understanding how motor circuits are organized and recruited in order to perform complex behavior is an essential question of neuroscience. Here we present an optogenetic protocol on larval zebrafish that allows spatial selective control of neuronal activity within a genetically defined population. We combine holographic illumination with the use of effective opsin transgenic lines, alongside high-speed behavioral monitoring to dissect the motor circuits of the larval zebrafish.

Speckle suppression in arbitrary parallel holographic illumination by the spatial frequency regaining method.

This Letter proposes a spatial frequency regaining method for parallel holographic illumination (SFR-PHI) to suppress speckle noise in phase-only computer-generated holography (CGH). Based on the accurate calculation of the beam bandwidth, this method uses the bandwidth-limited quadratic initial phase and weighted constraint iteration to generate the optimized phase hologram, which can provide the accurate spatial frequency of multiple illumination patterns. The results show that SFR-PHI performs superiorly in speckle suppression for generating dozens of illumination patterns in parallel and with arbitrary shapes and numbers. Compared with other speckle-suppression methods, it exhibits significant advantages in terms of accuracy and modulation efficiency.

24‐3: Distinguished Student Paper: Towards a Solid‐State LIDAR Using Holographic Illumination and a SPAD‐Based Time‐Of‐Flight Image Sensor

Light Detection and Ranging (LIDAR) systems are finding their way into an increasing number of applications including autonomous vehicles. Imaging based on Single Photon Avalanche Diode (SPAD) arrays has demonstrated great promise for LIDAR, typically using flood illumination or raster‐scanning of a small spot. Holographic illumination, on the other hand, has been successfully applied to display systems such as automotive Head Up Displays (HUDs). It holds promise as the method of illumination for a LIDAR system. We present a novel architecture and report initial results to show that it can improve the depth accuracy of 3D imaging.

Towards a solid‐state light detection and ranging system using holographic illumination and time‐of‐flight image sensing

AbstractLight detection and ranging (LIDAR) systems are finding their way into an increasing number of applications including autonomous vehicles. Image sensors based on single‐photon avalanche diode (SPAD) arrays have demonstrated great promise for LIDAR, typically using flood illumination or raster‐scanning of a small spot. Holographic illumination, based on phase‐only liquid crystal on silicon (LCoS) spatial light modulators, has been successfully applied to display systems such as automotive head up displays (HUDs). It holds great promise as the method of illumination for a LIDAR system. We present a novel solid‐state LIDAR architecture using holographic projection and single‐photon detection. We report a simple experiment that demonstrates proof of concept. The results of the experiment show that the novel architecture can improve the depth accuracy of 3D imaging compared to conventional LIDAR systems. We also indicate several key performance advantages of the novel architecture that will increase as the system scales to higher performance.

Enhanced Single-Beam Multiple-Intensity Phase Retrieval Using Holographic Illumination

Single-beam multiple-intensity iterative phase retrieval is a high-precision and lens-free computational imaging method, which reconstructs the complex-valued distribution of the object from a volume of axially captured diffraction intensities using the post-processing algorithm. However, for the object with slowly-varying waves, the method may encounter the problem of convergence stagnation since the lack of diversity between the captured intensity patterns. In this paper, a novel technique to enhance phase retrieval using holographic illumination is proposed. One special computer-generated hologram is designed, which can generate multiple significantly different images at the required distances. The incident plane wave is firstly modulated by the hologram, and then the exit wave is used to illuminate the object. Benefitting from this holographic illumination, remarkable intensity changes in the given detector planes can be produced, which is conducive to fast and high-accuracy reconstruction. Simulation and optical experiments are performed to verify the feasibility of the proposed method.

AUTOMATED TICKER-SYSTEM OF REMOTE DISPLAY OF DATA FOR LED-BOARD IN BENCH TESTS OF INTERNAL COMBUSTION ENGINE

The problem of developing adaptive information output systems, synchronized with the process of the experiment is solved by existing methods of concentration and interactive presentation of information. The purpose of this study is to develop an LED-board with an automated ticker-system for prompt display of information during bench tests of internal combustion engines. The developed design is based on the MAX7219 microcircuit. Data transfer occurs using the UART interface via the Bluetooth module HC-06. The driver is controlled via the SPI interface (mode 0). The choice of LED display is due to the possibility of using monochromatic light emitting diodes as point sources of radiation with a wide range of options for regulation of luminescence parameters. This is achieved when using high-performance LED clusters Pure-Green (525 nm), Ultra-Red (660 nm), Blue (470 nm). In the article the authors developed a constructive scheme of LED board for displaying the data of bench tests of internal combustion engines. The principle of the moving series (ticker), which is widely used in stock exchange analytics and advertising, is informative enough to reproduce the results of the internal combustion engine test bench tests. It has been determined that the application of the Arduino modular system will allow to vary the properties of the data display system to a wide extent and, when developing special software, to conduct test analytics using adaptive neural networks. The method of analysis and display of bench test data of an internal combustion engine has been improved. The developed scheme can be developed for holographic illumination of critical load zones on the surface of the structure under study. The slightly lower interactivity of holograms compared to augmented reality makes their supporting role more appropriate. It can be the basis for the formation on such a surface of markers for interactive augmented reality systems.

Holographic optics for thin and lightweight virtual reality

We present a class of display designs combining holographic optics, directional backlighting, laser illumination, and polarization-based optical folding to achieve thin, lightweight, and high performance near-eye displays for virtual reality. Several design alternatives are proposed, compared, and experimentally validated as prototypes. Using only thin, flat films as optical components, we demonstrate VR displays with thicknesses of less than 9 mm, fields of view of over 90° horizontally, and form factors approaching sunglasses. In a benchtop form factor, we also demonstrate a full color display using wavelength-multiplexed holographic lenses that uses laser illumination to provide a large gamut and highly saturated color. We show experimentally that our designs support resolutions expected of modern VR headsets and can scale to human visual acuity limits. Current limitations are identified, and we discuss challenges to obtain full practicality.

A compact holographic projector module for high-resolution 3D multi-site two-photon photostimulation.

Patterned two-photon (2P) photolysis via holographic illumination is a powerful method to investigate neuronal function because of its capability to emulate multiple synaptic inputs in three dimensions (3D) simultaneously. However, like any optical system, holographic projectors have a finite space-bandwidth product that restricts the spatial range of patterned illumination or field-of-view (FOV) for a desired resolution. Such trade-off between holographic FOV and resolution restricts the coverage within a limited domain of the neuron’s dendritic tree to perform highly resolved patterned 2P photolysis on individual spines. Here, we integrate a holographic projector into a commercial 2P galvanometer-based 2D scanning microscope with an uncaging unit and extend the accessible holographic FOV by using the galvanometer scanning mirrors to reposition the holographic FOV arbitrarily across the imaging FOV. The projector system utilizes the microscope’s built-in imaging functions. Stimulation positions can be selected from within an acquired 3D image stack (the volume-of-interest, VOI) and the holographic projector then generates 3D illumination patterns with multiple uncaging foci. The imaging FOV of our system is 800×800 μm2 within which a holographic VOI of 70×70×70 μm3 can be chosen at arbitrary positions and also moved during experiments without moving the sample. We describe the design and alignment protocol as well as the custom software plugin that controls the 3D positioning of stimulation sites. We demonstrate the neurobiological application of the system by simultaneously uncaging glutamate at multiple spines within dendritic domains and consequently observing summation of postsynaptic potentials at the soma, eventually resulting in action potentials. At the same time, it is possible to perform two-photon Ca2+ imaging in 2D in the dendrite and thus to monitor synaptic Ca2+ entry in selected spines and also local regenerative events such as dendritic action potentials.

Optogenetic Stimulation of Human Neural Networks Using Fast Ferroelectric Spatial Light Modulator—Based Holographic Illumination

The generation and application of human stem-cell-derived functional neural circuits promises novel insights into neurodegenerative diseases. These networks are often studied using stem-cell derived random neural networks in vitro, with electrical stimulation and recording using multielectrode arrays. However, the impulse response function of networks is best obtained with spatiotemporally well-defined stimuli, which electrical stimulation does not provide. Optogenetics allows for the functional control of genetically altered cells with light stimuli at high spatiotemporal resolution. Current optogenetic investigations of neural networks are often conducted using full field illumination, potentially masking important functional information. This can be avoided using holographically shaped illumination. In this article, we present a digital holographic illumination setup with a spatial resolution of about 8 µm, which suffices for the stimulation of single neurons, and offers a temporal resolution of less than 0.6 ms. With this setup, we present preliminary single-cell stimulation recording of stem-cell derived induced human neurons in a random neural network. This will offer the opportunity for further studies on connectivity in such networks.

Temperature Rise under Two-Photon Optogenetic Brain Stimulation

In recent decades, optogenetics has been transforming neuroscience research, enabling neuroscientists to drive and read neural circuits. The recent development in illumination approaches combined with two-photon (2P) excitation, either sequential or parallel, has opened the route for brain circuit manipulation with single-cell resolution and millisecond temporal precision. Yet, the high excitation power required for multi-target photostimulation, especially under 2P illumination, raises questions about the induced local heating inside samples. Here, we present and experimentally validate a theoretical model that makes it possible to simulate 3D light propagation and heat diffusion in optically scattering samples athigh spatial and temporal resolution under the illumination configurations most commonly used to perform 2P optogenetics: single- and multi-spot holographic illumination and spiral laser scanning. By investigating the effects of photostimulation repetition rate, spot spacing, and illumination dependence of heat diffusion, we found conditions that make it possible to design a multi-target 2P optogenetics experiment with minimal sample heating.

Two-Photon Holographic Stimulation of ReaChR.

Optogenetics provides a unique approach to remotely manipulate brain activity with light. Reaching the degree of spatiotemporal control necessary to dissect the role of individual cells in neuronal networks, some of which reside deep in the brain, requires joint progress in opsin engineering and light sculpting methods. Here we investigate for the first time two-photon stimulation of the red-shifted opsin ReaChR. We use two-photon (2P) holographic illumination to control the activation of individually chosen neurons expressing ReaChR in acute brain slices. We demonstrated reliable action potential generation in ReaChR-expressing neurons and studied holographic 2P-evoked spiking performances depending on illumination power and pulse width using an amplified laser and a standard femtosecond Ti:Sapphire oscillator laser. These findings provide detailed knowledge of ReaChR's behavior under 2P illumination paving the way for achieving in depth remote control of multiple cells with high spatiotemporal resolution deep within scattering tissue.

Computer Generated Holography with Intensity-Graded Patterns.

Computer Generated Holography achieves patterned illumination at the sample plane through phase modulation of the laser beam at the objective back aperture. This is obtained by using liquid crystal-based spatial light modulators (LC-SLMs), which modulate the spatial phase of the incident laser beam. A variety of algorithms is employed to calculate the phase modulation masks addressed to the LC-SLM. These algorithms range from simple gratings-and-lenses to generate multiple diffraction-limited spots, to iterative Fourier-transform algorithms capable of generating arbitrary illumination shapes perfectly tailored on the base of the target contour. Applications for holographic light patterning include multi-trap optical tweezers, patterned voltage imaging and optical control of neuronal excitation using uncaging or optogenetics. These past implementations of computer generated holography used binary input profile to generate binary light distribution at the sample plane. Here we demonstrate that using graded input sources, enables generating intensity graded light patterns and extend the range of application of holographic light illumination. At first, we use intensity-graded holograms to compensate for LC-SLM position dependent diffraction efficiency or sample fluorescence inhomogeneity. Finally we show that intensity-graded holography can be used to equalize photo evoked currents from cells expressing different levels of chanelrhodopsin2 (ChR2), one of the most commonly used optogenetics light gated channels, taking into account the non-linear dependence of channel opening on incident light.

Talbot multi-focal holographic fluorescence endoscopy for optically sectioned imaging.

A wide-field multi-plane endoscopic system incorporating multiplexed volume holographic gratings and Talbot illumination to simultaneously acquire optically sectioned fluorescence images of tissue structures from different depths is presented. The proposed endoscopic system is configured such that multiple Talbot-illumination planes occur inside a volumetric sample and serve as the input focal planes for the subsequent multiplexed volume holographic imaging gratings. We describe the design, implementation, and experimental data demonstrating this endoscopic system's ability to obtain optically sectioned multi-plane fluorescent images of tissue samples in wide-field fashion without scanning in lateral and axial directions.

Extended field-of-view and increased-signal 3D holographic illumination with time-division multiplexing.

Phase spatial light modulators (SLMs) are widely used for generating multifocal three-dimensional (3D) illumination patterns, but these are limited to a field of view constrained by the pixel count or size of the SLM. Further, with two-photon SLM-based excitation, increasing the number of focal spots penalizes the total signal linearly--requiring more laser power than is available or can be tolerated by the sample. Here we analyze and demonstrate a method of using galvanometer mirrors to time-sequentially reposition multiple 3D holograms, both extending the field of view and increasing the total time-averaged two-photon signal. We apply our approach to 3D two-photon in vivo neuronal calcium imaging.

Dynamic placement of plasmonic hotspots for super-resolution surface-enhanced Raman scattering.

In this paper, we demonstrate dynamic placement of locally enhanced plasmonic fields using holographic laser illumination of a silver nanohole array. To visualize these focused "hotspots", the silver surface was coated with various biological samples for surface-enhanced Raman spectroscopy (SERS) imaging. Due to the large field enhancements, blinking behavior of the SERS hotspots was observed and processed using a stochastic optical reconstruction microscopy algorithm enabling super-resolution localization of the hotspots to within 10 nm. These hotspots were then shifted across the surface in subwavelength (<100 nm for a wavelength of 660 nm) steps using holographic illumination from a spatial light modulator. This created a dynamic imaging and sensing surface, whereas static illumination would only have produced stationary hotspots. Using this technique, we also show that such subwavelength shifting and localization of plasmonic hotspots has potential for imaging applications. Interestingly, illuminating the surface with randomly shifting SERS hotspots was sufficient to completely fill in a wide field of view for super-resolution chemical imaging.

Material response of photopolymer containing four different photosensitizers

Building on a previous studies, the behaviour of four different photosensitizers in an acrylamide/polyvinyl alcohol (AA/PVA) photopolymer material are examined using a 1-D Nonlocal Photo-polymerisation Driven Diffusion (NPDD) model. In order to characterise the effects of using different photosensitizers, holographic illuminations with different spatial frequencies and intensities are applied. Material parameters, i.e., the nonlocal response parameter, σ, the diffusion rate of monomer, Dm, the chain initiation kinetic constant, ki, and the termination rate, kt, are extracted by numerically fitting the predictions of the NPDD to experimentally measured refractive index modulation growth curves. Four photosensitizers, [Erythrosin B (EB); Eosin Y (EY); Phloxine B (PB); and Rose Bengal (RB)], are examined.

Double-exposure phase calculation method in electronic speckle pattern interferometry based on holographic object illumination

Multiple-exposure phase calculation procedures are widely used in electronic speckle pattern interferometry to calculate phase maps of displacements. We developed a double-exposure process based on holographic illumination of the object and the idea of the spatial carrier phase-shifting method to examine transient displacements. In our work, computer-generated holograms and a spatial light modulator were used to generate proper coherent illuminating masks. In this adjustment all phase-shifted states were at our disposal from one recorded speckle image for phase calculation. This technique can be used in the large scale of transient measurements. In this paper we illustrate the principle through several examples.

Comparative electronic speckle pattern interferometry using adaptive holographic illumination

The development of spatial light modulators enables the application of active holographic optical elements in electronic speckle pattern interferometry. In our work adaptive comparative measurement is done, where an optically reconstructed image of a recorded or simulated single or double exposure hologram is used for holographic illumination of another object. In this paper, we present experimental results of measuring the difference of two deformations using this technique. The displacement difference can also be obtained numerically, if the wavefront used as a coherent illuminating mask does not belong to an existing object. This type of interferometer can easily adapt to the change of measuring conditions.