Kohler Illumination Research Articles

Large-range high-precision macroscopic common phase detection of segmented mirrors based on white light dynamic Twyman-Green interferometer

Segmented mirrors (SMs) provide a solution for the construction of the next generation of extremely large telescopes. The large-range, high-precision detection of piston error between mirror segments is a must-be solved problem in the application of SMs. This paper proposes a large-range high-precision macroscopic co-phase detection method of SMs based on white light dynamic Twyman-Green interferometer. By adopting a white light interferometry in the macroscopic co-phase detection, the probability of the occurring of the 2π ambiguity anomalies can be reduced. A piston error calculation model for SMs was established through Zernike fitting analysis, which improved the detection accuracy. A white light Twyman-Green interferometer with Kohler illumination was experimentally constructed. Based on the scanning of the reference mirror, an efficient detection of the piston error was achieved simultaneously with large range and high precision. Through the new method, the piston error of two planar mirrors was compensated from 38.9344 μm in coarse co-phase stage to 3.6 nm in fine co-phase stage. The method shows great potential for efficient co-phase adjustment of large primary segmented mirrors.

Irradiance-tailoring integral-illumination polarization homogenizer based on anamorphic aspheric microlens arrays.

In the realm of active polarization detection systems, the imperative for polarization illumination systems with high-uniformity and predefined-shape irradiance distribution is evident. This paper introduces a novel anamorphic aspheric (AAS) microlens array (MLA) integral polarization homogenizer, incorporating projection MLA (PMLA), condenser MLA (CMLA), polarization film (PF), and a sub-image array (SIA) mask based on Kohler illumination principles. Firstly, the optimal design of an AAS-based projection sub-lens is proposed to facilitate the creation of a short-working-distance, predefined-geometric and sharp polarization irradiance tailoring. The SIA mask is constituted by plenty of predistortion SIs, which are generated through a combination of chief ray tracing and the radial basis function (RBF) image warping method. In addition, accompanied with tolerance sensitivity analysis, detailed analysis of stray light generation factors and proposed elimination or suppression methods further ensure the engineering reliability and stability of the proposed system. A compact integral-illumination polarization homogenizer design example is realized with an overall irradiance uniformity exceeding 90% and a volume of 25 mm × 25 mm × 18.25 mm. Different predefined-geometrical-profile and high-uniformity polarization irradiance distribution can be achieved by substituting different SIA masks and PFs, without replacing MLA optical elements, which greatly saves cost. Substantial simulations and experiments corroborate the efficacy of our polarization homogenizer.

Design of large zoom ratio compact microscope based on coaxial Kohler illumination

Design of large zoom ratio compact microscope based on coaxial Kohler illumination

Design method of dual-band synchronous zoom microscope optical system based on coaxial Kohler illumination

The intrinsic properties of the observed object are closely related to its spectral information, to extend the imaging spectral range of a continuous zoom microscope to obtain more detailed intrinsic properties of the object, this paper proposes a design method of dual-band simultaneous zoom microscope optical system based on the coaxial Koehler uniform illumination. First, the imaging principle of the dual-band simultaneous zoom microscope optical system is theoretically analyzed, and we propose to split the front fixed group of the zoom system into a collimation lens group and a converging lens group to realize the compact design of the system. Then, two different rear fixed groups are used to correct the residual aberration, and a method for solving the initial structure of the dual-band simultaneous zoom microscope optical system is proposed. Finally, a dual-band synchronous zoom microscope optical system is designed using the method proposed in this paper. The design results show that the imaging magnification of the visible (VIS) band is −0.4 to −4.0, the simultaneous imaging magnification ranges are −0.4 to −0.8 in the VIS and short-wave infrared (SWIR) bands, and the magnification difference of its simultaneous zoom imaging is less than 1.25%. In addition, the system has the advantages of good imaging quality, clever design of coaxial illumination, and compact structure, thus verifying the feasibility of the design method.

Scalable Optical Convolutional Neural Networks Based on Free-Space Optics Using Lens Arrays and a Spatial Light Modulator.

A scalable optical convolutional neural network (SOCNN) based on free-space optics and Koehler illumination was proposed to address the limitations of the previous 4f correlator system. Unlike Abbe illumination, Koehler illumination provides more uniform illumination and reduces crosstalk. The SOCNN allows for scaling of the input array and the use of incoherent light sources. Hence, the problems associated with 4f correlator systems can be avoided. We analyzed the limitations in scaling the kernel size and parallel throughput and found that the SOCNN can offer a multilayer convolutional neural network with massive optical parallelism.

Experimental and numerical polarization analysis of the 3D transfer behavior in microsphere-assisted interferometry for 1D phase gratings

Enhancing the lateral resolution in optical microscopy and interferometry is of great interest in recent research. In order to laterally resolve structures including feature dimensions below the Abbe resolution limit, microspheres in the optical near-field of the specimen are shown to locally improve the resolution of the imaging system. Experimental and simulated results following this approach are obtained by a high NA Linnik interferometer and analyzed in this contribution. They show the reconstructed surface of a 1D phase grating below the resolution limit. For further understanding of the transfer characteristics, measured interference data are compared with FEM (finite element method) based simulations with respect to the polarization dependency of the relevant image information for 1D phase gratings. Therefore, the implemented Koehler illumination as well as the experimental setup utilize polarized light.

A scalable optical computer based on free-space optics using lens arrays and a spatial light modulator

A scalable optical computer based on free-space optics and Koehler illumination was proposed to obtain optical parallelism in an artificial neural network. The Koehler illumination scheme can provide more uniform illumination and less crosstalk compared with the previous systems based on Abbe illumination. The proposed design was realised in hardware with 2 × 2 inputs and 2 × 2 outputs using light-emitting diodes, a liquid crystal display, lens arrays, and a detector array assembled in an optical cage system with subsidiary electronics. The feasibility of the constructed optical computer was demonstrated by achieving the expected results of the NAND and NOR logics in parallel in the nondifference mode. Additionally, a simple pattern-checking function in the difference mode was shown. In addition, the limit of the scalability and the effect of geometric aberration was discussed. Furthermore, a clustering technique was suggested to overcome the limitations of scalability. Clustering can continue to increase the array size and throughput by creating a new block of doubled input and output arrays from multistacks of basic optical modules. The proposed architecture and clustering method have the potential to realise massive optical parallelism if large-array components and high-speed electronics are used in the future.

Optical Phase Contrast Microscopy with Incoherent Vortex Phase

AbstractA novel, compact, and broadband optical phase contrast microscopy based on incoherent vortex topological quadrupole is theoretically proposed and experimentally realized. The topological quadrupole, generated in a thin uniaxial crystal, possesses four single‐charge optical vortexes, each of which can act as a two‐dimensional (2D) optical spatial differentiator. The incoherence of light‐emitting diode (LED) light will wash out the optical vortex and the spatial differentiation effect, which can be ingeniously overcome by choosing Kohler illumination and inserting a 1 mm‐thickness uniaxial crystal sandwiched with two polarizers before the camera. By adjusting the polarizers and the tilted angle of crystal to selectively utilize the geometric Berry phase and the angular gradient of Fresnel transmission coefficients of crystal, the 1D, 2D, and second‐order analog spatial differentiations with a spatial resolution better than 0.775 μm can be switched flexibly. A versatile phase contrast microscope is built, which is well compatible with the conventional microscopes. The uniformly incoherent illumination without laser speckle effects results in high‐quality spatial differentiation imaging. Biologically transparent living cells are thus imaged with high contrast.

Three-dimensional transfer function of optical microscopes in reflection mode.

Three-dimensional (3D) transfer functions build the basis for a comprehensive characterization of optical imaging systems in the spatial frequency domain. Utilizing the projection-slice theorem, the 2D modulation transfer function of an incoherent imaging system can be derived from a 3D transfer function by integration with respect to the axial spatial frequency. For a diffraction limited microscope with homogeneous incoherent pupil illumination, the modulation transfer function equals the 2D autocorrelation function of a circular disc. However, until now to the best of our knowledge no 3D transfer function has been published, which exactly leads to the 2D modulation transfer function of a diffraction limited microscope in reflection mode. In this article, we derive a formula, which after integration with respect to the axial spatial frequency coordinate perfectly fits to the diffraction limited 2D modulation transfer function. The inverse three-dimensional Fourier transform of the 3D transfer function results in a complex-valued 3D point spread function, from which the depth of field, the lateral resolution and, in addition, the corresponding 3D point spread function of both, a conventional and an interference microscope, can beobtained.

Compact integrator design for short-distance sharp and unconventional geometric irradiance tailoring.

A compact microlens array (MLA) integral homogenizer composed of a projection MLA, a condenser MLA, and a subimage array mask based on Kohler illumination is presented herein. By adopting the optimal design of an aspheric projection sublens, a short-distance integrator for unconventional geometric irradiance tailoring can be acquired. Compared with the traditional integrator, the integral lens is removed in the proposed integrator. An incident beam with a larger divergence angle is permitted while a sharp illumination performance is maintained. In addition, the illumination distribution with a predefined geometrical profile can be obtained by introducing a subimage array mask without replacing the MLA elements. Through the introduced mask, the illumination attenuation area is cut, the distortion is caused by the large NA, and the peak effect caused by the integral lens of the traditional integrator can be eliminated. The subimage array mask is generated by combining ray tracing and the radial basis function image warping method. By introducing the array mask into the system, an 80% edge relative illumination and an 85% overall illumination uniformity are realized for a compact large-NA integrator with ${\rm NA} = {0.3}$ and thickness of 4.5 mm. An 88% edge relative illumination and a 92% overall illumination uniformity are realized for a small-NA integrator with ${\rm NA} = {0.15}$.

Brightness calculation formula for Kohler illumination beam

A two-lens optical system entails two beams: a crossover beam and a Kohler illumination beam. We observed that upon varying the excitation corresponding to the first lens from large to small values, the crossover beam changed to a Kohler illumination beam with increased brightness. We derived new calculation formulas for the Kohler illumination beam. Brightness B and beam current Ib can be expressed as B = Bco (ϕco/ϕ)2 and Ib = Bco (παϕco)2/4, where Bco, ϕco, ϕ, and α denote the crossover beam brightness, crossover beam size, Kohler illumination beam size, and beam semi-angle, respectively; additionally, ϕco and ϕ are obtained at the same first lens excitation. These two equations were experimentally validated. We obtained a brightness of 2.05 × 108 A/cm2 sr using a beam energy and an emission current of 20 keV and 54 µA, respectively. Notably, this value surpasses the Langmuir limit of 4.11 × 105 A/cm2 sr by 499 times.

4f Koehler Transmitted Illumination Condenser for Teaching and Low-Cost Microscopic Imaging

ABSTRACT Transmitted light imaging is an important tool in biophysics for applications that include sample analysis, recording samples whose viability is compromised by high levels of illumination (e.g., live cell tracking), and finding regions of interest in a sample. Koehler transillumination is a powerful illumination method used in commercial microscopes; yet commercial Koehler condensers are expensive, are difficult to integrate into tabletop systems, and make learning the concepts of Koehler illumination difficult because of their closed-box nature. Here, we show a protocol to build a simple 4f Koehler illumination system that offers advantages with respect to commercial condensers in terms of simplicity, cost, and compatibility with tabletop systems, such as open-source light sheet fluorescence microscopes. We include step-by-step instructions that can be followed by advanced undergraduate or graduate students without experience in optics on how to align and assemble the illuminator, along with a list of the necessary parts for assembly. We also include supplemental material that describes 4 supporting educational activities students can conduct with the apparatus and helps in the understanding of key concepts relevant to Koehler illumination and optics. The performance of the system is comparable to that of commercial condensers and significantly better, in terms of illumination homogeneity and depth of field (optical sections are possible), than that of LED flashlights, such as those found in low-cost diagnostic devices and tabletop systems.

Fundamentals of Microscopy.

The light (or optical) microscope is the icon of science. The aphorism "seeing is believing" is often quoted in scientific papers involving microscopy. Unlike many scientific instruments, the light microscope will deliver an image however badly it is set up. Fluorescence microscopy is a widely used research tool across all disciplines of biological and biomedical science. Most universities and research institutions have microscopes, including confocal microscopes. This introductory paper in a series detailing advanced light microscopy techniques explains the foundations of both electron and light microscopy for biologists and life scientists working with the mouse. An explanation is given of how an image is formed. A description is given of how to set up a light microscope, whether it be a brightfield light microscope on the laboratory bench, a widefield fluorescence microscope, or a confocal microscope. These explanations are accompanied by operational protocols. A full explanation on how to set up and adjust a microscope according to the principles of Köhler illumination is given. The importance of Nyquist sampling is discussed. Guidelines are given on how to choose the best microscope to image the particular sample or slide preparation that you are working with. These are the basic principles of microscopy that a researcher must have an understanding of when operating core bioimaging facility instruments, in order to collect high-quality images. © 2020 The Authors. Basic Protocol 1: Setting up Köhler illumination for a brightfield microscope Basic Protocol 2: Aligning the fluorescence bulb and setting up Köhler illumination for a widefield fluorescence microscope Basic Protocol 3: Generic protocol for operating a confocal microscope.

랜덤 포레스트 기반의 오토포커싱 알고리즘 개발

Auto-focusing is essential for the detection of minute defects in inspection equipments. We propose an optical system that can autofocus on a glass substrate and an algorithm that can adjust the focus quickly and accurately. The optical system uses Koehler illumination to measure the contrast of the field of view aperture, which is effective for plain glass. The auto-focusing algorithm using random forest is learned by setting the target value for the input image with the estimated focal length. When a new image is given, the focal length estimate is calculated. Various experiments have been performed on focus measurement using images acquired from various locations and show better results than others.

Design of UV LED illumination system for direct imaging lithography

UV LED, which can be used as the illumination source in the direct imaging (DI) exposure equipment, has the advantages of rich wavelengths and low cost. It has a good application prospect in the field of printed circuit board (PCB) manufacturing. In practical application, there are high requirements for output power density, stray light control, and luminance uniformity of DI system to improve the productivity and exposure performance. We present an optical structure of the illumination system for DI lithography to provide uniform illumination low stray light for digital micromirror device (DMD), which combines Kohler illumination and double telecentric imaging. In this design, the high luminance illumination can be obtained by LED etendue analysis, fly’s eye condenser is not needed, and the stray light on the DMD caused by light outside the effective angle of the LED can be mostly filtered out. Based on this idea, a lithographic illumination system with the numerical aperture of 0.1 is designed and fabricated for 0.95-in. DMD. According to experimental measurements, the effective illumination power is up to 10 W, the stray light accounts for 10.7%, and the illuminance uniformity is about 90%, which meet the requirements of DI lithography equipment used in PCB manufacturing.

Performing Spectroscopy on Plasmonic Nanoparticles with Transmission-Based Nomarski-Type Differential Interference Contrast Microscopy.

Differential interference contrast (DIC) microscopy is a powerful imaging tool that is most commonly employed for imaging microscale objects using visible-range light. The purpose of this protocol is to detail a proven method for preparing plasmonic nanoparticle samples and performing single particle spectroscopy on them with DIC microscopy. Several important steps must be followed carefully in order to perform repeatable spectroscopy experiments. First, landmarks can be etched into the sample substrate, which aids in locating the sample surface and in tracking the region of interest during experiments. Next, the substrate must be properly cleaned of debris and contaminants that can otherwise hinder or obscure examination of the sample. Once a sample is properly prepared, the optical path of the microscope must be aligned, using Kohler Illumination. With a standard Nomarski style DIC microscope, rotation of the sample may be necessary, particularly when the plasmonic nanoparticles exhibit orientation-dependent optical properties. Because DIC microscopy has two inherent orthogonal polarization fields, the wavelength-dependent DIC contrast pattern reveals the orientation of rod-shaped plasmonic nanoparticles. Finally, data acquisition and data analyses must be carefully performed. It is common to represent DIC-based spectroscopy data as a contrast value, but it is also possible to present it as intensity data. In this demonstration of DIC for single particle spectroscopy, the focus is on spherical and rod-shaped gold nanoparticles.

Quantitative Phase Imaging Camera With a Weak Diffuser

We introduce the quantitative phase imaging camera with a weak diffuser (QPICWD) as a practical realization of quantitative phase imaging (QPI) on standard microscope platforms. The QPICWD is a compact stand-alone camera which measures sample induced phase delay under low-coherence quasi-monochromatic illumination by examining the deformation of the speckle intensity pattern. By interpreting the speckle deformation with an ensemble average of the geometric flow, we can obtain the high-resolution distortion field by solving the transport of intensity equation (TIE). Since the phase measured by TIE is the generalized phase of the partially coherent image, instead of the phase of the measured object, we analyze the effect of illumination coherence and imaging numerical aperture (NA) on the accuracy of phase retrieval, revealing that the phase of the object can be reliably retrieved when the coherence parameter (the ratio of illumination NA to objective NA) of the Kohler illumination is between 0.3 and 0.5. We present several applications for the new design including nondestructive optical testing of microlens array with nanometric thickness and imaging of fixed and live HeLa cells. Since the designed QPI camera does not require any modification of the widely available bright-field microscope or additional accessories for its use, it is expected to be adopted by the broader biology and medical community.

Transform- and multi-domain deep learning for single-frame rapid autofocusing in whole slide imaging.

A whole slide imaging (WSI) system has recently been approved for primary diagnostic use in the US. The image quality and system throughput of WSI is largely determined by the autofocusing process. Traditional approaches acquire multiple images along the optical axis and maximize a figure of merit for autofocusing. Here we explore the use of deep convolution neural networks (CNNs) to predict the focal position of the acquired image without axial scanning. We investigate the autofocusing performance with three illumination settings: incoherent Kohler illumination, partially coherent illumination with two plane waves, and one-plane-wave illumination. We acquire ~130,000 images with different defocus distances as the training data set. Different defocus distances lead to different spatial features of the captured images. However, solely relying on the spatial information leads to a relatively bad performance of the autofocusing process. It is better to extract defocus features from transform domains of the acquired image. For incoherent illumination, the Fourier cutoff frequency is directly related to the defocus distance. Similarly, autocorrelation peaks are directly related to the defocus distance for two-plane-wave illumination. In our implementation, we use the spatial image, the Fourier spectrum, the autocorrelation of the spatial image, and combinations thereof as the inputs for the CNNs. We show that the information from the transform domains can improve the performance and robustness of the autofocusing process. The resulting focusing error is ~0.5 µm, which is within the 0.8-µm depth-of-field range. The reported approach requires little hardware modification for conventional WSI systems and the images can be captured on the fly without focus map surveying. It may find applications in WSI and time-lapse microscopy. The transform- and multi-domain approaches may also provide new insights for developing microscopy-related deep-learning networks. We have made our training and testing data set (~12 GB) open-source for the broad research community.

Using machine-learning to optimize phase contrast in a low-cost cellphone microscope.

Cellphones equipped with high-quality cameras and powerful CPUs as well as GPUs are widespread. This opens new prospects to use such existing computational and imaging resources to perform medical diagnosis in developing countries at a very low cost. Many relevant samples, like biological cells or waterborn parasites, are almost fully transparent. As they do not exhibit absorption, but alter the light’s phase only, they are almost invisible in brightfield microscopy. Expensive equipment and procedures for microscopic contrasting or sample staining often are not available. Dedicated illumination approaches, tailored to the sample under investigation help to boost the contrast. This is achieved by a programmable illumination source, which also allows to measure the phase gradient using the differential phase contrast (DPC) [1, 2] or even the quantitative phase using the derived qDPC approach [3]. By applying machine-learning techniques, such as a convolutional neural network (CNN), it is possible to learn a relationship between samples to be examined and its optimal light source shapes, in order to increase e.g. phase contrast, from a given dataset to enable real-time applications. For the experimental setup, we developed a 3D-printed smartphone microscope for less than 100 $ using off-the-shelf components only such as a low-cost video projector. The fully automated system assures true Koehler illumination with an LCD as the condenser aperture and a reversed smartphone lens as the microscope objective. We show that the effect of a varied light source shape, using the pre-trained CNN, does not only improve the phase contrast, but also the impression of an improvement in optical resolution without adding any special optics, as demonstrated by measurements.

Volume holographic imaging endoscopic design and construction techniques.

A reflectance volume holographic imaging (VHI) endoscope has been designed for simultaneous in vivo imaging of surface and subsurface tissue structures. Prior utilization of VHI systems has been limited to ex vivo tissue imaging. The VHI system presented in this work is designed for laparoscopic use. It consists of a probe section that relays light from the tissue sample to a handheld unit that contains the VHI microscope. The probe section is constructed from gradient index (GRIN) lenses that form a 1:1 relay for image collection. The probe has an outer diameter of 3.8 mm and is capable of achieving 228.1 ?? lp / mm resolution with 660-nm Kohler illumination. The handheld optical section operates with a magnification of 13.9 and a field of view of 390 ?? ? m × 244 ?? ? m . System performance is assessed through imaging of 1951 USAF resolution targets and soft tissue samples. The system has also passed sterilization procedures required for surgical use and has been used in two laparoscopic surgical procedures.