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 information. The principle has been experimentally validated by the lensless imaging of red blood cells of diameter ~6-9 micrometers and a photolithography mask with features in micrometer scale. The important advantages of the proposed approach of non-computational shadow microscopy are the improved depth of field and a drastic increase in the sensor to sample working distance. The imaging method can also be used as a projection technique in the multi-angle optical computed tomography (CT).
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…
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 …
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…
Lensless imaging is a popular research field for the advantages of small size, wide field-of-view and low aberration in recent years. However, some traditional lensless imaging methods suffer from slow convergence, mechanical errors and conjugate solution interference, which limit its further application and development. In this work, we proposed a lensless imaging method based on spatial light modulator (SLM) with unknown modulation curve. In our imaging system, we use SLM to modulate the wavefront of object, and introduce the ptychographic scanning algorithm that is able to recover the compl…
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…
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…
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…
Flexible fiber-optic endoscopes provide a minimally-invasive solution for imaging at depths beyond the reach of conventional microscopes. Current endoscopes require focusing and/or scanning mechanisms at the distal end, which limit miniaturization and frame-rate, and induce aberrations. Alternative lensless solutions are based on adaptive wavefront-correction, but are extremely sensitive to fiber bending. Here, we demonstrate a novel endoscopic approach, which enables single-shot imaging at a variable working distance through a conventional fiber bundle, without the use of any distal optics. O…
It is hard for us humans to recognize things in nature until we have invented them ourselves. For image-forming optics, nature has made virtually every kind of lens humans have devised. But what about lensless "imaging"? Recently, we showed that a bare array of sensors on a curved substrate could achieve resolution not limited by diffraction- without any lens at all provided that the objects imaged conform to our a priori assumptions. Is it possible that somewhere in nature we will find this kind of vision system? We think so and provide examples that seem to make no sense whatever unless they…
The pressing need for the detailed wavefront properties of ultra-bright and ultra-short pulses produced by free-electron lasers (FELs) has spurred the development of several complementary characterization approaches. Here we present a method based on ptychography that can retrieve full high-resolution complex-valued wave functions of individual pulses. Our technique is demonstrated within experimental conditions suited for diffraction experiments in their native imaging state. This lensless technique, applicable to many other short-pulse instruments, can achieve diffraction-limited resolution.
We describe a diffraction microscopy technique based on refractive optics to study structural variations in crystals. The X-ray beam diffracted by a crystal was magnified by beryllium parabolic refractive lenses on a 2D X-ray camera. The microscopy setup was integrated into the 6-circle Huber diffractometer at the ESRF beamline ID06. Our setup allowed us to visualize structural imperfections with a resolution of approximately 1 micrometer. The configuration, however, can easily be adapted for sub-micrometer resolution.
We present an analytical description and an experimental realization of interscale mixing microscopy, a diffraction-based imaging technique that is capable of detecting wavelength/10 objects in far-field measurements with both coherent and incoherent broadband light. This method aims at recovering the spatial spectrum of light diffracted by a subwavelength object based on far-field measurements of the interference created by the object and a finite diffraction grating. A single measurement, analyzing the multiple diffraction orders, is often sufficient to determine the parameters of the object…
Conventional image sensors are only responsive to the intensity variation of the incoming light wave. By encoding the wavefront information into the balanced detection scheme, we demonstrate an image sensor pixel design that is capable to detect both the local intensity and wavefront information simultaneously. With the full compatibility to the CMOS fabrication process, the proposed pixel design can benefit a variety of applications, including phase microscopy, lensless imaging and machine vision.
Conventional microscopic records represent intensity distributions whereby local sample information is mapped onto local information at the detector. In coherent microscopy, the superposition principle of waves holds; field amplitudes are added, not intensities. This non-local representation is spread out in space and interference information combined with wave continuity allows extrapolation beyond the actual detected data. Established resolution criteria are thus circumvented and hidden object details can retrospectively be recovered from just a fraction of an interference pattern.
In coherent X-ray diffraction microscopy the diffraction pattern generated by a sample illuminated with coherent x-rays is recorded, and a computer algorithm recovers the unmeasured phases to synthesize an image. By avoiding the use of a lens the resolution is limited, in principle, only by the largest scattering angles recorded. However, the imaging task is shifted from the experiment to the computer, and the algorithm's ability to recover meaningful images in the presence of noise and limited prior knowledge may produce aberrations in the reconstructed image. We analyze the low order aberrat…
Coherent X-ray diffraction microscopy is a method of imaging non-periodic isolated objects at resolutions only limited, in principle, by the largest scattering angles recorded. We demonstrate X-ray diffraction imaging with high resolution in all three dimensions, as determined by a quantitative analysis of the reconstructed volume images. These images are retrieved from the 3D diffraction data using no a priori knowledge about the shape or composition of the object, which has never before been demonstrated on a non-periodic object. We also construct 2D images of thick objects with infinite dep…
We present a theoretical investigation of coherent anti-Stokes Raman scattering (CARS) that is modulated by periodically depleting the ground state population through Rabi oscillations driven by an additional control laser. We find that such a process generates optical sidebands in the CARS spectrum and that the frequency of the sidebands depends on the intensity of the control laser light field. We show that analyzing the sideband frequency upon scanning the beams across the sample allows one to spatially resolve emitter positions where a spatial resolution of 65 nm, which is well below the d…
We report a computational 3D microscopy technique, termed Fourier ptychographic diffraction tomography (FPDT), that iteratively stitches together numerous variably illuminated, low-resolution images acquired with a low-numerical aperture (NA) objective in 3D Fourier space to create a wide field-of-view (FOV), high-resolution, depth-resolved complex refractive index (RI) image across large volumes. Unlike conventional optical diffraction tomography (ODT) approaches that rely on controlled bright-field illumination, holographic phase measurement, and high-NA objective detection, FPDT employs tom…
Light-field microscopy represents a promising solution for microscopic volumetric imaging, thanks to its capability to encode information on multiple planes in a single acquisition. This is achieved through its peculiar simultaneous capture of information on light spatial distribution and propagation direction. However, state-of-the-art light-field microscopes suffer from a detrimental loss of spatial resolution compared to standard microscopes. We propose and experimentally demonstrate a light-field microscopy architecture based on light intensity correlation, in which resolution is limited o…
Ptychography is an enabling coherent diffraction imaging technique for both fundamental and applied sciences. Its applications in optical microscopy, however, fall short for its low imaging throughput and limited resolution. Here, we report a resolution-enhanced parallel coded ptychography technique achieving the highest numerical aperture and an imaging throughput orders of magnitude greater than previous demonstrations. In this platform, we translate the samples across the disorder-engineered surfaces for lensless diffraction data acquisition. The engineered surface consists of chemically et…
The optical diffraction limit, formulated by Abbe 140 years ago, imposes a bound on imaging resolution in classical optics. Over the last twenty years, many theoretical schemes have been presented for overcoming the diffraction barrier in optical imaging using quantum properties of light. An experimental realization of sub-diffraction limited quantum imaging has, however, remained elusive. Here, we demonstrate a quantum imaging method taking advantage of non-classical light naturally produced in fluorescence microscopy due to photon antibunching, a fundamentally quantum phenomenon prohibiting …
Lensfree on-chip microscopy is an emerging imaging technique that can be used to visualize and study biological specimens without the need for imaging lens systems. Important issues that can limit the performance of lensfree on-chip microscopy include interferometric aberrations, acquisition noise, and image reconstruction artifacts. In this study, we introduce a Bayesian-based method for performing aberration correction and numerical diffraction that accounts for all three of these issues to improve the effective numerical aperture (NA) and signal-to-noise ratio (SNR) of the reconstructed mic…
Efficient imaging of biomolecules, 2D materials and electromagnetic fields depends on retrieval of the phase of transmitted electrons. We demonstrate a method to measure phase in a scanning transmission electron microscope using a nanofabricated diffraction grating to produce multiple probe beams. The measured phase is more interpretable than phase-contrast scanning transmission electron microscopy techniques without an off-axis reference wave, and the resolution could surpass that of off-axis electron holography. We apply the technique to image nanoparticles, carbon sub- strates and electric …
Speckle patterns have been widely used in imaging techniques such as ghost imaging, dynamic speckle illumination microscopy, structured illumination microscopy, and photoacoustic fluctuation imaging. Recent advances in the ability to control the statistical properties of speckles has enabled the customization of speckle patterns for specific imaging applications. In this work, we design and create special speckle patterns for parallelized nonlinear pattern-illumination microscopy based on fluorescence photoswitching. We present a proof-of-principle experimental demonstration where we obtain a …
I explain what is, what is not, and what is only sort of superresolution microscopy. I discuss optical resolution, first in terms of diffraction theory, then in terms of linear systems theory, and finally in terms of techniques that use prior information, nonlinearity, and other tricks to improve performance. The discussion reveals two classes of superresolution: Pseudo superresolution techniques improve images up to the diffraction limit but not much beyond. True superresolution techniques allow substantial, useful improvements beyond the diffraction limit. The two classes are distinguished b…
We report a novel extension to structured illumination (SI) microscopy that utilizes the Talbot self-imaging effect to generate a SI pattern on a sample with field-of-view (FOV) and resolution unconstrained by the numerical aperture (NA) of the microscope objective. This enables high-resolution SI pattern generation (up to the free-space propagation limit) across large FOVs, which in turn allows SI microscopy to be efficiently adapted for high-throughput imaging. We demonstrated this concept using a simple prototype microscope to image a large FOV (450 x 350 um) with a diffraction-limited fluo…
Scattering scanning near-field optical microscopy enables optical imaging and characterization of plasmonic devices with nanometer-scale resolution well below the diffraction limit. This technique enables developers to probe and understand the waveguide-coupled plasmonic antenna in as-fabricated heat-assisted magnetic recording heads. In order validate and predict results and to extract information from experimental measurements that is physically comparable to simulations, a model was developed to translate the simulated electric field into expected near-field measurements using physical para…
Acoustic-resolution optoacoustic microscopy (AR-OAM) retrieves anatomical and functional contrast from living tissues at depths not reachable with optical microscopy. The imaging performance of AR-OAM has been advanced with image reconstruction algorithms providing high lateral resolution ultimately limited by acoustic diffraction. In this work, we suggest a new model-based framework efficiently exploiting scanning symmetries for high-resolution reconstruction of AR-OAM images. The model accurately accounts for the spatial impulse response and large detection bandwidth of a spherical polyvinyl…
In this theoretical study, the author firstly discusses the wave interference of Bragg diffraction inside 3D crystal, followed by quantum mechanical interpretation on the diffraction process, and proves that the interference fringe between Bragg diffraction and the incident beam is identical to the lattice plane. By introducing the beam expander concept, we may explain the image projection mechanism of Bragg diffractions propagating through TEM: Transmission Electron Microscope. In practice, we will take projection images at zone axes, because of its symmetric high-density diffractions associa…
Light diffraction through a subwavelength aperture located at the apex of a metallic screen with conical geometry is investigated theoretically. A method based on a multipole field expansion is developed to solve Maxwell's equations analytically using boundary conditions adapted both for the conical geometry and for the finite conductivity of a real metal. The topological properties of the diffracted field are discussed in detail and compared to those of the field diffracted through a small aperture in a flat screen, i. e. the Bethe problem. The model is applied to coated, conically tapered op…
Deep Learning Enables Large Depth-of-Field Images for Sub-Diffraction-Limit Scanning Superlens Microscopy
City University of Hong Kong（香港城市大学）；State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences（中国科学院沈阳自动化研究所机器人学国家重点实验室）；Institutes for Robotics and Intelligent Manufacturing, Chinese Academy of Sciences（中国科学院机器人与智能制造创新研究院）；The University of Hong Kong（香港大学）；City University of Hong Kong Shenzhen Research Institute (CityUSRI)（香港城市大学深圳研究院）
Super-resolution imaging techniques enable nanoscale microscopy in fields such as physics, biology, and chemistry. However, many super-resolution techniques require specialized optical components, such as a helical-phase mask. We present a novel technique, Super-resolution Airy disk Microscopy (SAM) that can be used in a standard confocal microscope without any specialized optics. We demonstrate this technique, in combination with ground state depletion, to image and control nitrogen-vacancy (NV) centers in bulk diamond below the diffraction limit. A greater than 14-fold improvement in resolut…
One of the main characteristics of optical imaging systems is the spatial resolution, which is restricted by the diffraction limit to approximately half the wavelength of the incident light. Along with the recently developed classical super-resolution techniques, which aim at breaking the diffraction limit in classical systems, there is a class of quantum super-resolution techniques which leverage the non-classical nature of the optical signals radiated by quantum emitters, the so-called antibunching super-resolution microscopy. This approach can ensure a factor of $\sqrt{n}$ improvement in th…
Single-emitter microscopy has emerged as a promising method of imaging nanostructures with nanoscale resolution. This technique uses the centroid position of an emitters far-field radiation pattern to infer its position to a precision that is far below the diffraction limit. However, nanostructures composed of high-dielectric materials such as noble metals can distort the far-field radiation pattern. Nanoparticles also exhibit a more complex range of distortions, because in addition to introducing a high dielectric surface, they also act as efficient scatterers. Thus, the distortion effects of…
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.
Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the universal application of super-resolution microscopy is not feasible. In this paper, we propose and demonstrate a new kind of super-resolution fluorescence microscopy that can be easily implemented and requires neither additional hardware nor complex post-processing. The microscopy is based on the principle of stepwise optical saturation (SOS), where $M$ steps of raw fluoresce…
Modern optical microscopy methods have been advanced to provide super resolution at high imaging speed, but not chirality discriminative. We recently proposed chiral structured-illumination microscopy (SIM) method to image chiral fluorescent domains at sub-wavelength resolution. Chiral SIM is suitable for imaging chiral domains at sub-wavelength resolution but loses the high spatial frequency of the achiral ones. In order to obtain the full picture of all fluorescent domains at sub-wavelength resolution, we propose an advanced version of chiral SIM, termed double SIM, which enables simultaneou…
We demonstrate a three-dimensional (3D) optical diffraction tomographic technique with optimal frequency combination (OFC-ODT) for the 3D quantitative phase imaging of unlabeled specimens. Three sets of through-focus intensity images are captured under an annular aperture and two circular apertures with different coherence parameters. The 3D phase optical transfer functions (POTF) corresponding to different illumination apertures are combined to obtain an optimally synthesized frequency response, achieving high-quality, low-noise 3D reconstructions with imaging resolution up to the incoherent …
