We propose to use high numerical aperture single mode optical fibers like photonic crystal fiber for lensless in-line holographic microscopy. Highly divergent beam helps to overcome the spatial sampling limitation of the image sensor. In this paper, a submicron lateral resolution has been demonstrated, with an imaging sensor of pixel pitch 1.12 micrometer and a photonic crystal fiber of mode field diameter 1.8 micrometer. In earlier methods of single-shot lensless imaging, submicron resolution has been obtained at very small working distance and field of view. The proposed method improves the resolution without compromising the working distance. A working distance of (but not limited to) ~1.7 mm with a field of View ~1.4 mm has been demonstrated.
Recent development of lensless imagers has enabled three-dimensional (3D) imaging through a thin piece of optics in close proximity to a camera sensor. A general challenge of wide-field lensless imaging is the high computational complexity and slow speed to reconstruct 3D objects through iterative optimization process. Here, we demonstrated GEOMScope, a lensless 3D microscope that forms image through a single layer of microlens array and reconstructs objects through a geometrical-optics-based pixel back projection algorithm and background suppressions. Compared to others, our method allows loc…
This paper numerically investigates the feasibility of lensless zoomable holographic multiple projections to tilted planes. We have already developed lensless zoomable holographic single projection using scaled diffraction, which calculates diffraction between parallel planes with different sampling pitches. The structure of this zoomable holographic projection is very simple because it does not need a lens; however, it only projects a single image to a plane parallel to the hologram. The lensless zoomable holographic projection in this paper is capable of projecting multiple images onto tilte…
We report an angle-tilted, wavelength-multiplexed ptychographic modulation approach for multispectral lensless on-chip microscopy. In this approach, we illuminate the specimen with lights at 5 wavelengths simultaneously. A prism is added at the illumination path for spectral dispersion. Lightwaves at different wavelengths, thus, hit the specimen at slightly different incident angles, breaking the ambiguities in mixed state ptychographic reconstruction. At the detection path, we place a thin diffuser in-between the specimen and the monochromatic image sensor for encoding the spectral informatio…
This paper proposes a non-computational method of counteracting the effect of image degradation introduced by the diffraction phenomenon in lensless microscopy. All the optical images (whether focused by lenses or not) are diffraction patterns, which preserve the visual information upto a certain extent determined by the size of the point spread functions, like airy disks in some cases. A highly diverging beam can be exploited to reduce the spatial extent of these point spread functions relatively in the transformed projective space, which can help us in the spatial unmixing of the visual info…
Lensless microscopy with coherent or partially coherent light sources is a well known imaging technique, commonly referred as digital in-line holographic microscopy. In the established methods, both the spatial and temporal coherence of light play a crucial role in determining the resolution of reconstructed object. We report lensless microscopy with a spatially extended white LED, a light source of low spatial and very low temporal coherence. The wave-field propagation between two parallel planes can be obtained using a convolution operation, where the convolution kernel depends on the object…
On-chip holographic video is a convenient way to monitor biological samples simultaneously at high spatial resolution and over a wide field-of-view. However, due to the limited readout rate of digital detector arrays, one often faces a tradeoff between the per-frame pixel count and frame rate of the captured video. In this report, we propose a subsampled phase retrieval (SPR) algorithm to overcome the spatial-temporal trade-off in holographic video. Compared to traditional phase retrieval approaches, our SPR algorithm uses over an order of magnitude less pixel measurements while maintaining su…
We propose a novel framework for the systematic design of lensless imaging systems based on the hyperuniform random field solutions of nonlinear reaction-diffusion equations from pattern formation theory. Specifically, we introduce a new class of imaging point-spread-functions (PSFs) with enhanced isotropic behavior and controllable sparsity. We investigate the PSFs and the modulated transfer functions (MTFs) for a number of nonlinear models and demonstrate that two-phase isotropic random fields with hyperuniform disorder are ideally suited to construct imaging PSFs with improved performances …
We report a novel lensless on-chip microscopy platform based on near-field blind ptychographic modulation. In this platform, we place a thin diffuser in between the object and the image sensor for light wave modulation. By blindly scanning the unknown diffuser to different x-y positions, we acquire a sequence of modulated intensity images for quantitative object recovery. Different from previous ptychographic implementations, we employ a unit magnification configuration with a Fresnel number of ~50,000, which is orders of magnitude higher than previous ptychographic setups. The unit magnificat…
University of California, Berkeley（加州大学伯克利分校）；Orange Coast College（奥兰治海岸学院）
Compressive lensless imagers enable novel applications in an extremely compact device, requiring only a phase or amplitude mask placed close to the sensor. They have been demonstrated for 2D and 3D microscopy, single-shot video, and single-shot hyperspectral imaging; in each of these cases, a compressive-sensing-based inverse problem is solved in order to recover a 3D data-cube from a 2D measurement. Typically, this is accomplished using convex optimization and hand-picked priors. Alternatively, deep learning-based reconstruction methods offer the promise of better priors, but require many tho…
Lensless imaging is an elegant approach to high-resolution microscopy, which is rapidly gaining popularity in applications where imaging optics are problematic. However, current lensless imaging methods require objects to be placed within a well-defined support structure, while the light source needs to have a narrow, stable, and accurately known spectrum. Here we introduce a general approach to lensless imaging without spectral bandwidth limitations or sample requirements. We use two time-delayed coherent light pulses, and show that scanning the pulse-to-pulse time delay allows the reconstruc…
Fiber imaging bundles are widely used as thin, passive image conduits for miniaturised and endoscopic microscopy, particularly for confocal fluorescence imaging. Holographic microscopy through fiber bundles is more challenging; phase conjugation approaches are complex and require extensive calibration. This articles describes how simple inline holographic microscopy can be performed through an imaging bundle using a partially coherent illumination source from a multimode fiber. The sample is imaged in transmission, with the intensity hologram sampled by the bundle and transmitted to a remote c…
A holographic microscope captures interference patterns, or holograms, that encode three-dimensional (3D) information about the object being viewed. Computation is essential to extracting that 3D information. By wrapping low-level scattering codes and taking advantage of Python's data analysis ecosystem, HoloPy makes it easy for experimentalists to use modern, sophisticated inference methods to analyze holograms. The resulting data can be used to understand how small particles or microorganisms move and interact. The project illustrates how computational tools can enable experimental methods a…
Traditional microbial detection methods often rely on the overall property of microbial cultures and cannot resolve individual growth event at high spatiotemporal resolution. As a result, they require bacteria to grow to confluence and then interpret the results. Here, we demonstrate the application of an integrated ptychographic sensor for lensless cytometric analysis of microbial cultures over a large scale and with high spatiotemporal resolution. The reported device can be placed within a regular incubator or used as a standalone incubating unit for long-term microbial monitoring. For longi…
We propose a three-dimensional holographic reconstruction procedure applicable with no a priori knowledge about the recording conditions enabling distortion-free three-dimensional object reconstruction.
Holographic microscopy has developed into a powerful tool for 3D particle tracking, yielding nanometer-scale precision at high frame rates. However, current particle tracking algorithms ignore the effect of the microscope objective on the formation of the recorded hologram. As a result, particle tracking in holographic microscopy is currently limited to particles well above the microscope focus. Here, we show that modeling the effect of an aberration-free lens allows tracking of particles above, near, and below the focal plane in holographic microscopy, doubling the depth of field. Finally, we…
We have extended Laser Doppler holographic microscopy to transmission geometry. The technique is validated with living fish embryos imaged by a modified upright bio-microcope. By varying the frequency of the holographic reference beam, and the combination of frames used to calculate the hologram, multimodal imaging has been performed. Doppler images of the blood vessels for different Doppler shifts, images where the flow direction is coded in RGB colors or movies showing blood cells individual motion have been obtained as well. The ability to select the Fourier space zone that is used to calcu…
This paper describes an imaging microscopic technique based on heterodyne digital holography where subwavelength-sized gold colloids can be imaged in cell environment. Surface cellular receptors of 3T3 mouse fibroblasts are labeled with 40 nm gold nanoparticles, and the biological specimen is imaged in a total internal reflection configuration with holographic microscopy. Due to a higher scattering efficiency of the gold nanoparticles versus that of cellular structures, accurate localization of a gold marker is obtained within a 3D mapping of the entire sample's scattered field, with a lateral…
We use digital holographic microscopy and Mie scattering theory to simultaneously characterize and track individual colloidal particles. Each holographic snapshot provides enough information to measure a colloidal sphere's radius and refractive index to within 1%, and simultaneously to measure its three-dimensional position with nanometer in-plane precision and 10 nanometer axial resolution.
Using conventional refraction-based optical lens, it is challenging to achieve both high-resolution imaging and long-working-distance condition. To increase the numerical aperture of a lens, the working distance should be compensated, and vice versa. Here we propose and demonstrate a new concept in optical microscopy that can achieve both high-resolution imaging and long-working-distance conditions by utilising a scattering layer instead of refractive optics. When light passes through a scattering layer, it creates a unique interference pattern. To retrieve the complex amplitude image from the…
We present a new technique that combines off-axis Digital Holography and Dark Field Microscopy to track 100nm gold particles diffusing in water. We show that a single hologram is sufficient to localize several particles in a thick sample with a localization accuracy independent of the particle position. From our measurements we reconstruct the trajectories of the particles and derive their 3D diffusion coefficient. Our results pave the way for quantitative studies of the motion of single nanoparticle in complex media.
We report experimental results on heterodyne holographic microscopy of subwavelength-sized gold particles. The apparatus uses continuous green laser illumination of the metal beads in a total internal reflection configuration for dark-field operation. Detection of the scattered light at the illumination wavelength on a charge-coupled device array detector enables 3D localization of brownian particles in water
We present a cost-effective, simple and fast digital holographic microscopy method based upon Rayleigh-Sommerfeld back propagation for identification of the geometrical shape of a cell. The method was tested using synthetic hologram images generated by ray-tracing software and from experimental images of semi-transparent spherical beads and living red blood cells. Our results show that by only using the real part of the back-reconstructed amplitude the proposed method can provide information of the geometrical shape of the object and at the same time accurately determine the axial position of …
We demonstrate a gigapixel inline digital holographic microscopy using a consumer scanner. The consumer scanner can maximally scan an A4 size image (297mm x 210mm) with 4800 dpi (= 5.29 um), theoretically achieving a resolution of 56,144 x 39,698 = 2.22 gigapixels. The system using a consumer scanner has a simple structure, compared with synthetic aperture digital holography using a camera mounted on a two-dimensional moving stage. In this demonstration, we captured an inline hologram with 23,602 x 18,023 pixels (= 0.43 gigapixels). In addition, to accelerate the reconstruction time of the gig…
In this paper, we explore the 3D structure of light scattering from dark-field illuminated live 3T3 cells in the presence of 40 nm gold nanomarkers. For this purpose, we use a high resolution holographic microscope combining the off-axis heterodyne geometry and the phase-shifting acquisition of the digital holograms. A comparative study of the 3D reconstructions of the scattered fields allows us to locate the gold markers which yield, contrarily to the cell structures, well defined bright scattering patterns that are not angularly titled and clearly located along the optical axis (z). This cha…
We propose a real-time digital holographic microscopy, that enables simultaneous multiple reconstructed images with arbitrary resolution, depth and positions, using Shifted-Fresnel diffraction instead of Fresnel diffraction. In this system, we used four graphics processing units (GPU) for multiple reconstructions in real-time. We show the demonstration of four reconstruction images from a hologram with arbitrary depths, positions, and resolutions.
This study developed handheld and low-cost digital holographic microscopy (DHM) by adopting an in-line type hologram, a webcam, a high power RGB light emitting diode (LED), and a pinhole. It cost less than 20,000 yen (approximately 250 US dollars at 80 yen/dollar), and was approximately 120 mm x 80 mm x 55 mm in size. In addition, by adjusting the recording-distance of a hologram, the lateral resolution power at the most suitable distance was 17.5 um. Furthermore, this DHM was developed for use in open source libraries, and is therefore low-cost and can be easily developed by anyone. In this r…
Phase retrieval and imaging phase measurements are fields of intense research. It has recently been shown that phase retrieval from self-referencing interferograms (SRI) can be reformulated leading to a stable, linear equation provided the amplitude of the field is known from prior measurement steps (HOLOCAM). Consequently, the numerical solution thereof is straightforward. This is a big achievement since convergence is otherwise not always guaranteed. Applications are expected in X-ray microscopy, general phase retrieval, holography, tomography and optical imaging.
University of California, Los Angeles（加州大学洛杉矶分校）；California NanoSystems Institute（加州纳米系统研究所）；David Geffen School of Medicine（大卫·格芬医学院）
Polarized light microscopy provides high contrast to birefringent specimen and is widely used as a diagnostic tool in pathology. However, polarization microscopy systems typically operate by analyzing images collected from two or more light paths in different states of polarization, which lead to relatively complex optical designs, high system costs or experienced technicians being required. Here, we present a deep learning-based holographic polarization microscope that is capable of obtaining quantitative birefringence retardance and orientation information of specimen from a phase recovered …
Laboratoire Kastler Brossel（卡斯特勒·布罗塞尔实验室）；Ecole Normale Supérieure（巴黎高等师范学院）；UPMC（巴黎第六大学（皮埃尔和玛丽·居里大学））；Laboratoire Charles Coulomb（查尔斯·库仑实验室）；Université Montpellier II（蒙彼利埃第二大学）
In this paper we present a digital holographic microscope to track gold colloids in three dimensions. We report observations of 100nm gold particles in motion in water. The expected signal and the chosen method of reconstruction are described. We also discuss about how to implement the numerical calculation to reach real-time 3D tracking.
An holographic microscopy reconstruction method compatible with high numerical aperture microscope objective (MO) up to NA=1.4 is proposed. After off axis and reference field curvature corrections, and after selection of the +1 grating order holographic image, a phase mask that transforms the optical elements of the holographic setup into an afocal device is applied in the camera plane. The reconstruction is then made by the angular spectrum method. The field is first propagated in the image half space from the camera to the afocal image of the MO optimal plane (plane for which MO has been des…
This Letter reports a demonstration of off-axis compressed holography in low-light level imaging conditions. An acquisition protocol relying on a single exposure of a randomly undersampled diffraction map of the optical field, recorded in the high heterodyne gain regime, is proposed. The image acquisition scheme is based on compressed sensing, a theory establishing that near-exact recovery of an unknown sparse signal is possible from a small number of nonstructured measurements. Image reconstruction is further enhanced by introducing an off-axis spatial support constraint to the image estimati…
Usually, off-axis digital holographic microscopy requires a coherent light source in order to record a full-field hologram. Nevertheless, a LASER-based illumination leads to a non-negligible coherent noise, decreasing then the imaging quality. We hereby report a simple method to reduce the coherent noise contribution using a low-temporal-coherence illumination while maintaining a large interference area. A diffraction grating is hence introduced in the reference arm of the interferometer, allowing the coherence plane of the reference beam to be tilted following angular dispersion. The phase pl…
This paper presents an application of digital Fourier holography for selective imaging of scatterers with different sizes in turbid media such as biological tissues. A combination of Fourier holography and high-resolution digital recording, digital Fourier microscopy (DFM) permits crucial flexibility in applying filtering to highlight scatterers of interest in the tissue. The high-resolution digital hologram is a result of the collation of Fourier holographic frames to form a large-size composite hologram. It is expected that DFM has an improved signal-to-noise ratio as compared to conventiona…
