Untrained deep learning-based differential phase-contrast microscopy.

Quantitative differential phase-contrast (DPC) microscopy produces phase images of transparent objects based on a number of intensity images. To reconstruct the phase, in DPC microscopy, a linearized model for weakly scattering objects is considered; this limits the range of objects to be imaged, and requires additional measurements and complicated algorithms to correct for system aberrations. Here, we present a self-calibrated DPC microscope using an untrained neural network (UNN), which incorporates the nonlinear image formation model. Our method alleviates the restrictions on the object to be imaged and simultaneously reconstructs the complex object information and aberrations, without any training dataset. We demonstrate the viability of UNN-DPC microscopy through both numerical simulations and LED microscope-based experiments.

Full Stokes polarimetry using a monolithic off-axis polarizing interferometer and a 2D array sensor.

This paper describes a full Stokes polarimeter employing a monolithic off-axis polarizing interferometric module and a 2D array sensor. The proposed passive polarimeter provides a dynamic full Stokes vector measurement capability of around 30 Hz. As the proposed polarimeter employs no active devices and is operated by employing an imaging sensor, it has significant potential to become a highly compact polarization sensor for smartphone applications. To show the feasibility of the proposed passive dynamic polarimeter scheme, the full Stokes parameters of a quarter-wave plate are extracted and displayed on a Poincare sphere by varying the polarization state of the measured beam.

Engineering pupil function for optical adversarial attacks.

Adversarial attacks inject imperceptible noise to images to deteriorate the performance of deep image classification models. However, most of the existing studies consider attacks in the digital (pixel) domain where an image acquired by an image sensor with sampling and quantization is recorded. This paper, for the first time, introduces a scheme for optical adversarial attack, which physically alters the light field information arriving at the image sensor so that the classification model yields misclassification. We modulate the phase of the light in the Fourier domain using a spatial light modulator placed in the photographic system. The operative parameters of the modulator for adversarial attack are obtained by gradient-based optimization to maximize cross-entropy and minimize distortion. Experiments based on both simulation and a real optical system demonstrate the feasibility of the proposed optical attack. We show that our attack can conceal perturbations in the image more effectively than the existing pixel-domain attack. It is also verified that the proposed attack is completely different from common optical aberrations such as spherical aberration, defocus, and astigmatism in terms of both perturbation patterns and classification results.

Dynamic spectroscopic imaging ellipsometry.

A dynamic spectroscopic imaging ellipsometer (DSIE) employing a monolithic polarizing interferometer is described. The proposed DSIE system can provide spatio-spectral ellipsometric phase map data Δ(λ, x) dynamically at a speed of 30 Hz. We demonstrate the ultrafast mapping capability of the spectroscopic ellipsometer by measuring a patterned 8-inch full wafer with a spatial resolution of less than 50 × 50 µm2 in an hour.

Two concepts for ultra-high-resolution polarization-sensitive optical coherence tomography with a single camera.

Two designs with a multiplexed superluminescent diode for ultra-high-resolution spectral-domain polarization-sensitive optical coherence tomography (UHR-PS-OCT) are introduced. In the first design, a Wollaston prism separates orthogonal polarization states next to each other on one linescan camera; the other design uses a beam displacer to separate orthogonal states onto two lines of a linescan camera with multiple rows of detectors. The coherence lengths measured with the two systems were 3.6 µm and 2.9 µm (n=1.38), respectively. Measurements were collected from the fovea of a healthy subject, a healthy subject with astigmatism, and a patient with central serous retinopathy (CSR). A single volumetric scan provides double pass retardance induced by the retinal nerve fiber layer birefringence (RNFL) and its birefringence, the cumulative double pass retardance induced by the Henle fiber layer, and the retardance that is induced by the retinal pigment epithelium-Bruch's membrane complex. The high axial resolution in UHR-PS-OCT is particularly helpful for the measurements of thin retinal tissue, such as the RNFL in the fovea, where birefringence values of around 1°/µm were found. Tilting of the retina due to a CSR or by off centering the imaging beam in the pupil causes an artificial increase in the double pass retardance induced by the RNFL and Henle fiber layer.

Polarization-sensitive differential phase-contrast microscopy.

We present a novel, to the best of our knowledge, form of polarization microscopy capable of producing quantitative optic-axis and phase retardation maps of transparent and anisotropic materials. The proposed method operates on differential phase-contrast (DPC) microscopy that produces a phase image of a thin specimen using multi-axis intensity measurements. For polarization-sensitive imaging, patterned illumination light is circularly polarized to illuminate a specimen. The light transmitted through a specimen is split into two orthogonal polarization states and measured by an image sensor. Subsequent DPC computation based on the illumination patterns, acquired images, and the imaging model enables the retrieval of polarization-dependent quantitative phase images, which are utilized to reconstruct the orientation and retardation of the specimen. We demonstrate the validity of the proposed method by measuring the optic-axis and phase retardation maps of calibrated and various anisotropic samples.

Direct replication of a glass micro Fresnel zone plate array by laser irradiation using an infrared transmissive mold.

It is not yet possible to fabricate micrometer-scale, glass optical components with nanometer-scale precision. Glass thermal imprinting enhances production efficiency. However, dimensional changes caused by shrinkage are inevitable because of phase transitions. Replication is very difficult when high-level pitch precision is essential. We used an infrared-transparent silicon mold and a CO2 laser to perform replica-type, thermal surface texturing at the nanoscale level; we analyzed a glass Fresnel zone plate array to this end. The Fresnel zone plate array was 10 × 10 mm2 in area and featured a 20 × 20 array. The individual Fresnel zone plate diameter was 500 µm and had 21 rings of minimum linewidth 2.9 µm and height 737 nm.

Single-pixel, single-input-state polarization-sensitive wavefront imaging.

In this Letter, we describe a single-pixel polarization-sensitive imaging technique, capable of generating the birefringence map of a thin specimen by using single-pixel detectors. Spatially modulated light is circularly polarized to illuminate the specimen. The transmitted light through the specimen is then focused via a lens and measured by position-sensitive detectors in two orthogonal polarization channels. The measurement of the irradiance and centroid position of the optical focus and subsequent computations enable the production of polarization-dependent wavefront maps, which can then be utilized to reconstruct sample birefringence information. We demonstrate the feasibility of our method by measuring distribution of optic-axis orientation and phase retardation of various birefringent samples.

Color-coded LED microscopy for quantitative phase imaging: Implementation and application to sperm motility analysis.

Color-coded light-emitting diode (LED) microscopy (cLEDscope) is a novel computational microscopy technique capable of multi-contrast and quantitative phase imaging of biological specimens using color-multiplexed illumination. Using specially designed LED patterns, it is capable of recording multiple differential phase contrast (DPC) images in a single exposure and employs a computational algorithm to retrieve the phase distribution of the specimens. Herein, we describe the detailed procedures in the cLEDscope implementation for quantitative phase imaging. Several notable features and caveats in the cLEDscope setup and image processing are also outlined. The imaging model is derived for our specific configuration, and the associated phase-retrieval algorithms are presented on the basis of a weak-object transfer function. As an illustrative application of the quantitative cLEDscope, we demonstrate its utility as a sperm-motility analyzer by exploiting its real-time quantitative imaging capability.

Miniaturized Sample Preparation and Rapid Detection of Arsenite in Contaminated Soil Using a Smartphone.

Conventional methods for analyzing heavy metal contamination in soil and water generally require laboratory equipped instruments, complex procedures, skilled personnel and a significant amount of time. With the advancement in computing and multitasking performances, smartphone-based sensors potentially allow the transition of the laboratory-based analytical processes to field applicable, simple methods. In the present work, we demonstrate the novel miniaturized setup for simultaneous sample preparation and smartphone-based optical sensing of arsenic As(III) in the contaminated soil. Colorimetric detection protocol utilizing aptamers, gold nanoparticles and NaCl have been optimized and tested on the PDMS-chip to obtain the high sensitivity with the limit of detection of 0.71 ppm (in the sample) and a correlation coefficient of 0.98. The performance of the device is further demonstrated through the comparative analysis of arsenic-spiked soil samples with standard laboratory method, and a good agreement with a correlation coefficient of 0.9917 and the average difference of 0.37 ppm, are experimentally achieved. With the android application on the device to run the experiment, the whole process from sample preparation to detection is completed within 3 hours without the necessity of skilled personnel. The approximate cost of setup is estimated around 1 USD, weight 55 g. Therefore, the presented method offers the simple, rapid, portable and cost-effective means for onsite sensing of arsenic in soil. Combined with the geometric information inside the smartphones, the system will allow the monitoring of the contamination status of soils in a nation-wide manner.

Design of binary phase filters for depth-of-focus extension via binarization of axisymmetric aberrations.

We present a novel design approach for a binary phase mask with depth-of-focus (DoF) extension ability. Our method considers that the binarized version of an axisymmetric continuous phase pupil generates twin-intensity profiles that are symmetric with respect to the focal plane, each of which resembles the focal behavior of its continuous original. The DoF extension is realized by repositioning and coherently summing the twin foci to achieve an elongated focus along the axial direction. The shift of the two foci towards the focal plane can be handled by superimposing the defocus term in the continuous pupil function. We demonstrate our proposed design approach for two representative axisymmetric aberration functions, i.e., defocused phase axicon and spherical aberration. The manipulation of topological parameters in the phase axicon and spherical aberration, along with the defocus strength, enables the multiple binary phase-filter designs of DoF extension of 3.2-7.1 fold with a phase axicon and 2.8-14.8 fold with a spherical aberration, compared to the case with a clear aperture.

Single-exposure quantitative phase imaging in color-coded LED microscopy.

We demonstrate single-shot quantitative phase imaging (QPI) in a platform of color-coded LED microscopy (cLEDscope). The light source in a conventional microscope is replaced by a circular LED pattern that is trisected into subregions with equal area, assigned to red, green, and blue colors. Image acquisition with a color image sensor and subsequent computation based on weak object transfer functions allow for the QPI of a transparent specimen. We also provide a correction method for color-leakage, which may be encountered in implementing our method with consumer-grade LEDs and image sensors. Most commercially available LEDs and image sensors do not provide spectrally isolated emissions and pixel responses, generating significant error in phase estimation in our method. We describe the correction scheme for this color-leakage issue, and demonstrate improved phase measurement accuracy. The computational model and single-exposure QPI capability of our method are presented by showing images of calibrated phase samples and cellular specimens.

Frequency- and spectrally-encoded confocal microscopy.

We describe a three-dimensional microscopy technique based on spectral and frequency encoding. The method employs a wavelength-swept laser to illuminate a specimen with a spectrally-dispersed line focus that sweeps over the specimen in time. The spatial information along each spectral line is further mapped into different modulation frequencies. Spectrally-resolved detection and subsequent Fourier analysis of the back-scattered light from the specimen therefore enable high-speed, scanner-free imaging of the specimen with a single-element photodetector. High-contrast, three-dimensional imaging capability of this method is demonstrated by presenting images of various materials and biological specimens.

Dark-field polarization-sensitive optical coherence tomography.

Polarization-sensitive optical coherence tomography (PS-OCT) is a functional OCT providing both structural and birefringent information of the sample, and it has been applied to the studies of various organs having polarization properties. Fiber-based PS-OCT is sensitive to specular reflection from the sample surface, because signal saturation due to the strong specular reflection can make the polarization measurement difficult. We developed a dark-field PS-OCT which can avoid the specular reflection problem. Dark-field PS-OCT was implemented by adapting a hybrid method of Bessel-beam illumination and Gaussian-beam detection, and a PS-OCT method based on passive delay unit (PDU). The new system was characterized in comparison with the conventional Gaussian-beam based method in both polarization components and various samples including the human skin. Dark-field PS-OCT performed as good as the conventional PS-OCT without the specular reflection artifact. Dark-field PS-OCT may be useful in practical situations where the specular reflection is unavoidable.

Microfluidic sorting of fluorescently activated cells depending on gene expression level.

During the last few years, fluorescence activated cell sorter has played an important role in a variety of biological investigations as well as clinical diagnostics. However, the conventional fluorescence activated cell sorter has several limitations, such as large size, large sample volumes required for operation, and high cost. In this paper, we present a novel microfluidic device that can separate cells based on various fluorescent protein expression levels. Our system consists of three major parts: focusing, detection, and separation. The operating principles are briefly as follows: first fluorescent cells were delivered into the microfluidic chip and focused in the center of channel by sheath flow. Subsequently, the cells were excited by a 532 nm laser at 30 µW and concurrently detected by a photomultiplier tube. Based on their fluorescence intensities, the cells were separated into three outlets by a dielectrophoretic force. Using this system, we successfully separated the genetically modified cells at 0.1 µL/min (sample flow rate) to sheath flow rate at 1:5, 5 Vpp voltage, and 800 kHz frequency. The separation efficiency was measured as high as 94.7%. In conclusion, we found that our system has the capability of separating genetically modified cells with various fluorescent intensities and help study biology and medicine in a molecular level.

Detection of pH-induced aggregation of "smart" gold nanoparticles with photothermal optical coherence tomography.

We report the feasibility of a novel contrast agent, namely "smart" gold nanoparticles (AuNPs), in the detection of cancer cells with photothermal optical coherence tomography (PT-OCT). "Smart" AuNPs form aggregation in low pH condition, which is typical for cancer cells, and this aggregation results in a shift of their absorption spectrum. A PT-OCT system was developed to detect this pH-induced aggregation by combining an OCT light source and a laser with 660 nm in wavelength for photothermal excitation. Optical detection of pH-induced aggregation was tested with solution samples at two different pH conditions. An increase in optical path length (OPL) variation was measured at mild acidic condition, while there was not much change at neutral condition. Detection of cancer cells was tested with cultured cell samples. HeLa and fibroblast cells, as cancer and normal cells respectively, were incubated with "smart" gold nanoparticles and measured with PT-OCT. An elevated OPL variation signal was detected with the HeLa cells while not much of a signal was detected with the fibroblast cells. With the novel optical property of "smart" AuNPs and high sensitivity of PT-OCT, this technique is promising for cancer cell detection.

Field-based dynamic light scattering microscopy: theory and numerical analysis.

We present a theoretical framework for field-based dynamic light scattering microscopy based on a spectral-domain optical coherence phase microscopy (SD-OCPM) platform. SD-OCPM is an interferometric microscope capable of quantitative measurement of amplitude and phase of scattered light with high phase stability. Field-based dynamic light scattering (F-DLS) analysis allows for direct evaluation of complex-valued field autocorrelation function and measurement of localized diffusive and directional dynamic properties of biological and material samples with high spatial resolution. In order to gain insight into the information provided by F-DLS microscopy, theoretical and numerical analyses are performed to evaluate the effect of numerical aperture of the imaging optics. We demonstrate that sharp focusing of fields affects the measured diffusive and transport velocity, which leads to smaller values for the dynamic properties in the sample. An approach for accurately determining the dynamic properties of the samples is discussed.

Non-Invasive Retinal Blood Vessel Wall Measurements with Polarization-Sensitive Optical Coherence Tomography for Diabetes Assessment: A Quantitative Study.

Diabetes affects the structure of the blood vessel walls. Since the blood vessel walls are made of birefringent organized tissue, any change or damage to this organization can be evaluated using polarization-sensitive optical coherence tomography (PS-OCT). In this paper, we used PS-OCT along with the blood vessel wall birefringence index (BBI = thickness/birefringence2) to non-invasively assess the structural integrity of the human retinal blood vessel walls in patients with diabetes and compared the results to those of healthy subjects. PS-OCT measurements revealed that blood vessel walls of diabetic patients exhibit a much higher birefringence while having the same wall thickness and therefore lower BBI values. Applying BBI to diagnose diabetes demonstrated high accuracy (93%), sensitivity (93%) and specificity (93%). PS-OCT measurements can quantify small changes in the polarization properties of retinal vessel walls associated with diabetes, which provides researchers with a new imaging tool to determine the effects of exercise, medication, and alternative diets on the development of diabetes.

Polarization-sensitive intensity diffraction tomography.

Optical anisotropy, which is an intrinsic property of many materials, originates from the structural arrangement of molecular structures, and to date, various polarization-sensitive imaging (PSI) methods have been developed to investigate the nature of anisotropic materials. In particular, the recently developed tomographic PSI technologies enable the investigation of anisotropic materials through volumetric mappings of the anisotropy distribution of these materials. However, these reported methods mostly operate on a single scattering model, and are thus not suitable for three-dimensional (3D) PSI imaging of multiple scattering samples. Here, we present a novel reference-free 3D polarization-sensitive computational imaging technique-polarization-sensitive intensity diffraction tomography (PS-IDT)-that enables the reconstruction of 3D anisotropy distribution of both weakly and multiple scattering specimens from multiple intensity-only measurements. A 3D anisotropic object is illuminated by circularly polarized plane waves at various illumination angles to encode the isotropic and anisotropic structural information into 2D intensity information. These information are then recorded separately through two orthogonal analyzer states, and a 3D Jones matrix is iteratively reconstructed based on the vectorial multi-slice beam propagation model and gradient descent method. We demonstrate the 3D anisotropy imaging capabilities of PS-IDT by presenting 3D anisotropy maps of various samples, including potato starch granules and tardigrade.

E2E-BPF microscope: extended depth-of-field microscopy using learning-based implementation of binary phase filter and image deconvolution.

Several image-based biomedical diagnoses require high-resolution imaging capabilities at large spatial scales. However, conventional microscopes exhibit an inherent trade-off between depth-of-field (DoF) and spatial resolution, and thus require objects to be refocused at each lateral location, which is time consuming. Here, we present a computational imaging platform, termed E2E-BPF microscope, which enables large-area, high-resolution imaging of large-scale objects without serial refocusing. This method involves a physics-incorporated, deep-learned design of binary phase filter (BPF) and jointly optimized deconvolution neural network, which altogether produces high-resolution, high-contrast images over extended depth ranges. We demonstrate the method through numerical simulations and experiments with fluorescently labeled beads, cells and tissue section, and present high-resolution imaging capability over a 15.5-fold larger DoF than the conventional microscope. Our method provides highly effective and scalable strategy for DoF-extended optical imaging system, and is expected to find numerous applications in rapid image-based diagnosis, optical vision, and metrology.