Intrasession repeatability and agreement of a new method to measure the foveal fixation axis.

Purpose: Ophthalmic lens adaptation, particularly with progressive addition lenses, requires accurate measurements of the patient nasopupillary distance (NPD) and interpupillary distance (IPD), which are usually collected using the pupil centre as a reference. However, differences between the pupil centre and visual or foveal axis could induce some subsidiary effects of correcting lenses. This study aimed to assess the intrasession repeatability of a new prototype (Ergofocus®; Lentitech, Barakaldo, Spain) that can measure the foveal fixation axis (FFA) distance and assess the agreement with the NPD measurements collected using a traditional method (frame ruler). Methods: The FFA at far and near distances was measured three consecutive times in 39 healthy volunteers to determine the intrasession repeatability according to the British Standards Institute and International Organization for Standardization. Additionally, the FFA and NPD (standard frame ruler) were measured in 71 healthy volunteers and compared using Bland-Altman analysis. Two blinded experienced practitioners conducted each FFA and NPD measurement. Results: The FFA measurements showed acceptable repeatability at far distances (right eye (RE): Sw = 1.16 ± 0.76 mm and coefficient of variation (CV) = 3.92 ± 2.51%; left eye (LE) Sw = 1.11 ± 0.79 mm and CV = 3.76 ± 2.51%) and at near distances (RE: Sw = 0.97 ± 0.85 mm and CV = 3.52 ± 3.02%; LE: Sw = 1.17 ± 0.96 mm and CV = 4.54 ± 3.72%). Additionally, agreement with the NPD showed large differences at far distances (RE: -2.15 ± 2.34, LoA = -6.73 to 2.43 mm (P < 0.001); LE: -0.61 ± 2.62, LoA = -5.75 to 4.53 mm (P = 0.052)) and near distances (RE: -3.08 ± 2.80, LoA -8.57 to 2.42 mm (P < 0.001); LE: -2.97 ± 3.97, LoA: -10.75 to 4.80 mm (P < 0.001)). Conclusions: FFA measurements showed clinically acceptable repeatability at both far and near distances. Agreement with the NPD measured using a standard frame ruler showed significant differences, suggesting that both measurements are not interchangeable in clinical practice to prescribe and center ophthalmic lenses. Further research is necessary to assess the impact of FFA measurement in ophthalmic lens prescriptions.

[Features of accumulation of chemical elements in the volume of the lens in senile cataract]. / Osobennosti nakopleniya khimicheskikh elementov v ob"eme khrustalika pri senil'noi katarakte.

PURPOSE: This study provides a detailed analysis of the bioinorganic chemical composition of lens substance in patients with senile cataract using classical and spatial statistics methods. MATERIAL AND METHODS: The study included 30 isolated human lenses. The light scattering ability (LSA) of the lens substance was evaluated using an original method. Additionally, distribution of chemical elements in the lens substance was analyzed using a scanning electron microscope with energy dispersive spectrometer (SEM/EDS). Measurements by all methods were carried out in a single coordinate space, which made it possible to compare the spatial correlation of different parameters. RESULTS: Small-angle light scattering of the lens substance has been quantitatively characterized for the first time. In contrast to the conventional norm, in senile cataract the accumulation fields of the majority of ion-forming elements (including Na, P, K, Cl) are distributed along the lines repeating the geometry of the lens capsule. At the same time, the light scattering ability of certain areas of the lens is significantly correlated with changes in the concentrations of Na, P, K, Ca in these areas. In particular, one ion-forming element can be distinguished - Na: spatial change of its concentration in senile cataract is strongly associated with a local change in LSA of the lens with opacities clustering of any degree. Thus, a change in the nature of the Na accumulation in the lens volume can be considered the main marker of senile cataract formation. CONCLUSION: The distribution pattern of ion-forming elements indicates that the loss of barrier properties in the capsule plays a significant role in the development of senile cataract.

Large field-of-view short-wave infrared metalens for scanning fiber endoscopy.

Significance: The scanning fiber endoscope (SFE), an ultrasmall optical imaging device with a large field-of-view (FOV) for having a clear forward view into the interior of blood vessels, has great potential in the cardiovascular disease diagnosis and surgery assistance, which is one of the key applications for short-wave infrared biomedical imaging. The state-of-the-art SFE system uses a miniaturized refractive spherical lens doublet for beam projection. A metalens is a promising alternative that can be made much thinner and has fewer off-axis aberrations than its refractive counterpart. Aim: We demonstrate a transmissive metalens working at 1310 nm for a forward viewing endoscope to achieve a shorter device length and better resolution at large field angles. Approach: We optimize the metalens of the SFE system using Zemax, fabricate it using e-beam lithography, characterize its optical performances, and compare them with the simulations. Results: The SFE system has a resolution of ∼ 140 µ m at the center of field (imaging distance 15 mm), an FOV of ∼ 70 deg , and a depth-of-focus of ∼ 15 mm , which are comparable with a state-of-the-art refractive lens SFE. The use of the metalens reduces the length of the optical track from 1.2 to 0.86 mm. The resolution of our metalens-based SFE drops by less than a factor of 2 at the edge of the FOV, whereas the refractive lens counterpart has a ∼ 3 times resolution degradation. Conclusions: These results show the promise of integrating a metalens into an endoscope for device minimization and optical performance improvement.

Multi-Illumination Single-Holographic-Exposure Lensless Fresnel (MISHELF) Microscopy: Principles and Biomedical Applications.

Lensless holographic microscopy (LHM) comes out as a promising label-free technique since it supplies high-quality imaging and adaptive magnification in a lens-free, compact and cost-effective way. Compact sizes and reduced prices of LHMs make them a perfect instrument for point-of-care diagnosis and increase their usability in limited-resource laboratories, remote areas, and poor countries. LHM can provide excellent intensity and phase imaging when the twin image is removed. In that sense, multi-illumination single-holographic-exposure lensless Fresnel (MISHELF) microscopy appears as a single-shot and phase-retrieved imaging technique employing multiple illumination/detection channels and a fast-iterative phase-retrieval algorithm. In this contribution, we review MISHELF microscopy through the description of the principles, the analysis of the performance, the presentation of the microscope prototypes and the inclusion of the main biomedical applications reported so far.

Biomimetic human eyes in adaptive lenses with conductive gels.

To investigate the imaging effect, adaptive robust lenses are prepared by sealing transparent liquid or gel. Lenses are fabricated using the negative-pressure method, which is a benefit for a stable biconvex shape. Under the action of an electric field, the soft lens deforms following the dielectric elastomer actuator (DEA). DE (dielectric elastomer) membranes expand in the plane perpendicular to the electric field lines. The toroidal driving area leads to a decrease in lens diameter and an increase in convex curvature. Therefore, the focal length of the lens becomes shorter. The experimental measurement utilizes the double focal length method. As a result, the largest focal length change that could be achieved was 44.7% (190 mm→105 mm) of the soft lens using a DEA with carbon grease electrodes. Furthermore, the ECG (electrocardiogram) conductive gel could replace traditional carbon grease for DEA electrodes in optics. This type of transparent electrode is creatively applied to a biomedical lens. Under the same conditions, the electrostriction rate in a DEA with ECG gel was achieved at 33%, which was greater than that of 28% in a DEA coupled with carbon grease electrode. Adaptive lenses have characteristics such as easy fabrication, low cost, and strong operability, and they possess great potential application value in biomedical feild.

Recent Progress and Applications of Thermal Lens Spectrometry and Photothermal Beam Deflection Techniques in Environmental Sensing.

This paper presents recent development and applications of thermal lens microscopy (TLM) and beam deflection spectrometry (BDS) for the analysis of water samples and sea ice. Coupling of TLM detection to a microfluidic system for flow injection analysis (µFIA) enables the detection of microcystin-LR in waters with a four samples/min throughput (in triplicate injections) and provides an LOD of 0.08 µg/L which is 12-times lower than the MCL for microcystin-LR in water. µFIA-TLM was also applied for the determination of total Fe and Fe(II) in 3 µL samples of synthetic cloudwater. The LODs were found to be 100 nM for Fe(II) and 70 nM for total Fe. The application of µFIA-TLM for the determination of ammonium in water resulted in an LOD of 2.3 µM for injection of a 5 µL sample and TLM detection in a 100 µm deep microfluidic channel. For the determination of iron species in sea ice, the BDS was coupled to a diffusive gradient in the thin film technique (DGT). The 2D distribution of Fe(II) and total Fe on DGT gels provided by the BDS (LOD of 50 nM) reflected the distribution of Fe species in sea ice put in contact with DGT gels.

Design of a digital PCR optical detection system with multiple fluorescent microdroplets.

The need for accurate nucleic acid testing is increasing with the spread of the global pandemic. Problems such as low efficiency and precision and large volume exist because the number of existing digital polymerase chain reaction (PCR) testing instrument channels is low. In order to solve these problems, a four channel micro drop digital PCR system was designed. The collimating lens set and objective lens of the LED light source are designed in ZEMAX software, and the entire optical path is simulated. It is verified that the light energy utilization rate of the system is over 93% and that stray light interference is marginal.

Bessel acoustic-beam acoustic lens for extending the depth of field of detection in optical-resolution photoacoustic microscopy.

As an important part of optical-resolution photoacoustic microscopy, the acoustic lens is responsible for efficient collection of photoacoustic signals. The spherical focused acoustic lens is commonly used in photoacoustic microscopy because of its efficient detection of the photoacoustic signal in the focus area. However, the narrow depth of field of the spherical focused acoustic lens limits the expansion of the depth of field of the photoacoustic microscopy. To solve this problem, a Bessel acoustic-beam acoustic lens is proposed. The Bessel acoustic-beam acoustic lens replaces the spherical concave surface with a conical concave surface to generate a Bessel acoustic beam with non-diffraction. Using the simulation model of Bessel acoustic-beam acoustic lens constructed by COMSOL Multiphysics, it is verified theoretically that the Bessel acoustic-beam acoustic lens can improve the depth of field of detection by ∼2 times. The Bessel acoustic-beam acoustic lens can further promote the capability of high-speed and large volumetric imaging of optical-resolution photoacoustic microscopy and will be helpful in the acquisition of physiological and pathological processes.

Simulation Analysis of a Wavefront Reconstruction of a Large Aperture Laser Beam.

In order to solve the problem of atmospheric influence on the far-field measurement of the quality of a laser beam, we proposed a direct wavefront measurement system based on the Hartmann detection principle, which can measure large apertures and high-power laser beams. The measuring system was composed of a lens array and a detector. The wavefront detection of a large aperture laser beam could be realized by controlling the distance between the lenses and the size of the lens. The influence of different duty cycle factors on the accuracy of the wavefront reconstruction under the same arrangement and different arrangement conditions was simulated and analyzed. The simulation results showed that when the sub-lenses of the system were not in close contact, the reconstruction accuracy of the duty factor of 0.8 was close to that of the case of the duty factor of 1. Within a certain detection range, the hexagonal arrangement of 19 lenses and the arrangement of 8 × 8 lens arrays had a high wavefront restoration accuracy; both were lower than 0.10 λ. The system proposed in this paper was suitable for measuring a large aperture laser beam, providing a new idea for measuring and analyzing the quality of large aperture laser beams. It also has an important significance for improving the measurement accuracy of the beam quality.

Dual-resonant scanning multiphoton microscope with ultrasound lens and resonant mirror for rapid volumetric imaging.

A dual-resonant scanning multiphoton (DRSM) microscope incorporating a tunable acoustic gradient index of refraction lens with a resonant mirror is developed for high-speed volumetric imaging. In the proposed microscope, the pulse train signal of a femtosecond laser is used to trigger an embedded field programmable gate array to sample the multiphoton excited fluorescence signal at the rate of one pixel per laser pulse. It is shown that a frame rate of around 8000 Hz can be obtained in the x-z plane for an image region with a size of 256 × 80 pixels. Moreover, a volumetric imaging rate of over 30 Hz can be obtained for a large image volume of 343 × 343 × 120 µm3 with an image size of 256 × 256 × 80 voxels. Moreover, a volumetric imaging rate of over 30 Hz can be obtained for a large image volume of 256 × 256 × 80 voxels, which represents 343 × 343 × 120 µm3 in field-of-view. The rapid volumetric imaging rate eliminates the aliasing effect for observed temporal frequencies lower than 15 Hz. The practical feasibility of the DRSM microscope is demonstrated by observing the mushroom bodies of a drosophila brain and performing 3D dynamic observations of moving 10-µm fluorescent beads.

Deblur or denoise: the role of an aperture in lens and neural network co-design.

Co-design methods have been introduced to jointly optimize various optical systems along with neural network processing. In the literature, the aperture is generally a fixed parameter although it controls an important trade-off between the depth of focus, the dynamic range, and the noise level in an image. In contrast, we include aperture in co-design by using a differentiable image formation pipeline that models the effect of the aperture on the image noise, dynamic, and blur. We validate this pipeline on examples of image restoration and extension of the depth of focus. These simple examples illustrate the importance of optimizing the aperture in the co-design framework.

Ultraportable Flow Cytometer Based on an All-Glass Microfluidic Chip.

The flow cytometer has become a powerful and widely accepted measurement device in both biological studies and clinical diagnostics. The application of the flow cytometer in emerging point-of-care scenarios, such as instant detection in remote areas and emergency diagnosis, requires a significant reduction in physical dimension, cost, and power consumption. This requirement promotes studies to develop portable flow cytometers, mostly based on the utilization of polymer microfluidic chips. However, due to the relatively poor optical performance of polymer materials, existing microfluidic flow cytometers are incapable of accurate blood analysis, such as the four-part leukocyte differential count, which is necessary to monitor the immune system and to assess the risk of allergic inflammation or viral infection. To address this issue, an ultraportable flow cytometer based on an all-glass microfluidic chip (AG-UFCM) has been developed in this study. Compared with that of a typical commercial flow cytometer (BD FACSAria III), the volume of the AG-UFCM was reduced by 90 times (from 720 to 8 L). A two-step laser processing was employed to fabricate an all-glass microfluidic chip with a surface roughness of less than 1 nm, significantly improving the optical performance of on-chip micro-lens. The signal-to-noise ratio was enhanced by 3 dB, compared with that of polymer materials. For the first time, a four-part leukocyte differential count based on single fluorescence staining was realized using a miniaturized flow cytometer, laying a foundation for the point-of-care testing of miniaturized flow cytometers.

Lenz Lenses in a Cryoprobe: Boosting NMR Sensitivity Toward Environmental Monitoring of Mass-Limited Samples.

Nuclear magnetic resonance (NMR) spectroscopy is commonly employed in a wide range of metabolomic research. Unfortunately, due to its relatively low sensitivity, smaller samples become challenging to study by NMR. Cryoprobes can be used to increase sensitivity by cooling the coil and preamplifier, offering sensitivity improvements of ∼3 to 4x. Alternatively, microcoils can be used to increase mass sensitivity by improving sample filling and proximity, along with decreased electrical resistance. Unfortunately, combining the two approaches is not just technically challenging, but as the coil decreases, so does its thermal fingerprint, reducing the advantage of cryogenic cooling. Here, an alternative solution is proposed in the form of a Lenz lens inside a cryoprobe. Rather than replacing the detection coil, Lenz lenses allow the B1 field from a larger coil to be refocused onto a much smaller sample area. In turn, the stronger B1 field at the sample provides strong coupling to the cryocoil, improving the signal. By combining a 530 I.D. Lenz lens with a cryoprobe, sensitivity was further improved by 2.8x and 3.5x for 1H and 13C, respectively, over the cryoprobe alone for small samples. Additionally, the broadband nature of the Lenz lenses allowed multiple nuclei to be studied and heteronuclear two-dimensional (2D) NMR approaches to be employed. The sensitivity improvements and 2D capabilities are demonstrated on 430 nL of hemolymph and eight eggs (∼350 µm O.D.) from the model organismDaphnia magna. In summary, combining Lenz lenses with cryoprobes offers a relatively simple approach to boost sensitivity for tiny samples while retaining cryoprobe advantages.

Near-real-time diagnosis of electron optical phase aberrations in scanning transmission electron microscopy using an artificial neural network.

The key to optimizing spatial resolution in a state-of-the-art scanning transmission electron microscope is the ability to measure and correct for electron optical aberrations of the probe-forming lenses precisely. Several diagnostic methods for aberration measurement and correction have been proposed, albeit often at the cost of relatively long acquisition times. Here, we illustrate how artificial intelligence can be used to provide near-real-time diagnosis of aberrations from individual Ronchigrams. The demonstrated speed of aberration measurement is important because microscope conditions can change rapidly. It is also important for the operation of MEMS-based hardware correction elements, which have less intrinsic stability than conventional electromagnetic lenses.

Estimation of Lens Stray Light with Regard to the Incapacitation of Imaging Sensors-Part 2: Validation.

Recently, we developed a simple theoretical model for the estimation of the irradiance distribution at the focal plane of commercial off-the-shelf (COTS) camera lenses in case of laser illumination. The purpose of such a model is to predict the incapacitation of imaging sensors when irradiated by laser light. The model is based on closed-form equations that comprise mainly standard parameters of the laser dazzle scenario and those of the main devices involved (laser source, camera lens and imaging sensor). However, the model also includes three non-standard parameters, which describe the scattering of light within the camera lens. In previous work, we have performed measurements to derive these typically unknown scatter parameters for a collection of camera lenses of the Double-Gauss type. In this publication, we compare calculations based on our theoretical model and the measured scatter parameters with the outcome of stray light simulations performed with the optical design software FRED in order to validate the reliability of our theoretical model and of the derived scatter parameters.

Simplified superoscillatory lenses for super-resolution imaging.

In recent years, superoscillations have become a new method for creating super-resolution imaging systems. The design of superoscillatory wavefronts and their corresponding lenses can, however, be a complicated process. In this study, we extend a recently developed method for designing complex superoscillatory filters to the creation of phase- and amplitude-only filters and compare their performance. These three types of filters can generate nearly identical superoscillatory fields at the image plane.

Uniformity Correction of CMOS Image Sensor Modules for Machine Vision Cameras.

Flat-field correction (FFC) is commonly used in image signal processing (ISP) to improve the uniformity of image sensor pixels. Image sensor nonuniformity and lens system characteristics have been known to be temperature-dependent. Some machine vision applications, such as visual odometry and single-pixel airborne object tracking, are extremely sensitive to pixel-to-pixel sensitivity variations. Numerous cameras, especially in the fields of infrared imaging and staring cameras, use multiple calibration images to correct for nonuniformities. This paper characterizes the temperature and analog gain dependence of the dark signal nonuniformity (DSNU) and photoresponse nonuniformity (PRNU) of two contemporary global shutter CMOS image sensors for machine vision applications. An optimized hardware architecture is proposed to compensate for nonuniformities, with optional parametric lens shading correction (LSC). Three different performance configurations are outlined for different application areas, costs, and power requirements. For most commercial applications, the correction of LSC suffices. For both DSNU and PRNU, compensation with one or multiple calibration images, captured at different gain and temperature settings are considered. For more demanding applications, the effectiveness, external memory bandwidth, power consumption, implementation, and calibration complexity, as well as the camera manufacturability of different nonuniformity correction approaches were compared.

A novel feature for monitoring the enzymatic harvesting process of adherent cell cultures based on lens-free imaging.

Adherent cell cultures are often dissociated from their culture vessel (and each other) through enzymatic harvesting, where the detachment response is monitored by an operator. However, this approach is lacking standardisation and reproducibility, and prolonged exposure or too high concentrations can affect the cell's viability and differentiation potential. Quantitative monitoring systems are required to characterise the cell detachment response and objectively determine the optimal time-point to inhibit the enzymatic reaction. State-of-the-art methodologies rely on bulky imaging systems and/or features (e.g. circularity) that lack robustness. In this study, lens-free imaging (LFI) technology was used to develop a novel cell detachment feature. Seven different donors were cultured and subsequently harvested with a (diluted) enzymatic harvesting solution after 3, 5 and 7 days of culture. Cell detachment was captured with the LFI set-up over a period of 20 min (every 20 s) and by optimising the reconstruction of the LFI intensity images, a new feature could be identified. Bright regions in the intensity image were identified as detaching cells and using image analysis, a method was developed to automatically extract this feature, defined as the percentage of detached cell regions. Next, the method was quantitatively and qualitatively validated on a diverse set of images. Average absolute error values of 1.49%, 1.34% and 1.97% were obtained for medium to high density and overconfluent cultures, respectively. The detachment response was quantified for all conditions and the optimal time for enzyme inhibition was reached when approximately 92.5% of the cells were detached. On average, inhibition times of 9.6-11.1 and 16.2-17.2 min were obtained for medium to high density and overconfluent cultures, respectively. In general, overconfluent cultures detached much slower, while their detachment rate was also decreased by the diluted harvesting solution. Moreover, several donors exhibited similar trends in cell detachment behaviour, with two clear outliers. Using the novel feature, measurements can be performed with an increased robustness, while the compact LFI design could pave the way for in situ monitoring in a variety of culture vessels, including bioreactors.