Subwavelength Resolution Fourier Ptychography with Hemispherical Digital Condensers
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Introduction to Light Microscopy
Introduction to light microscopy A CAMDU training course Claire Mitchell, Imaging specialist, L1.01, 08-10-2018 Contents 1.Introduction to light microscopy 2.Different types of microscope 3.Fluorescence techniques 4.Acquiring quantitative microscopy data 1. Introduction to light microscopy 1.1 Light and its properties 1.2 A simple microscope 1.3 The resolution limit 1.1 Light and its properties 1.1.1 What is light? An electromagnetic wave A massless particle AND γ commons.wikimedia.org/wiki/File:EM-Wave.gif www.particlezoo.net 1.1.2 Properties of waves Light waves are transverse waves – they oscillate orthogonally to the direction of propagation Important properties of light: wavelength, frequency, speed, amplitude, phase, polarisation upload.wikimedia.org 1.1.3 The electromagnetic spectrum 퐸푝ℎ표푡표푛 = ℎν 푐 = λν 퐸푝ℎ표푡표푛 = photon energy ℎ = Planck’s constant ν = frequency 푐 = speed of light λ = wavelength pion.cz/en/article/electromagnetic-spectrum 1.1.4 Refraction Light bends when it encounters a change in refractive index e.g. air to glass www.thetastesf.com files.askiitians.com hyperphysics.phy-astr.gsu.edu/hbase/Sound/imgsou/refr.gif 1.1.5 Diffraction Light waves spread out when they encounter an aperture. electron6.phys.utk.edu/light/1/Diffraction.htm The smaller the aperture, the larger the spread of light. 1.1.6 Interference When waves overlap, they add together in a process called interference. peak + peak = 2 x peak constructive trough + trough = 2 x trough peak + trough = 0 destructive www.acs.psu.edu/drussell/demos/superposition/superposition.html 1.2 A simple microscope 1.2.1 Using lenses for refraction 1 1 1 푣 = + 푚 = physicsclassroom.com 푓 푢 푣 푢 cdn.education.com/files/ Light bends as it encounters each air/glass interface of a lens. -
The-Pathologists-Microscope.Pdf
The Pathologist’s Microscope The Pathologist’s Microscope Rudolf Virchow, the father of Pathology, had available to him wonderful microscopes during the 1850’s to 1880’s, but the one you have now is far better. Your microscope is the most highly perfected of all scientific instruments. These brief notes on alignment, the objective lens, the condenser, and the eyepieces are what you need to know to get the most out of your microscope and to feel comfortable using it. Figure 1 illustrates the important parts of a generic modern light microscope. Figure 1 - Parts of the Microscope UNC Pathology & Lab Med, MSL, July 2013 1 The Pathologist’s Microscope Alignment August Köhler, in 1870, invented the method for aligning the microscope’s optical system that is still used in all modern microscopes. To get the most from your microscope it should be Köhler aligned. Here is how: 1. Focus a specimen slide at 10X. 2. Open the field iris and the condenser iris. 3. Observe the specimen and close the field iris until its shadow appears on the specimen. 4. Use the condenser focus knob to bring the field iris into focus on the specimen. Try for as sharp an image of the iris as you can get. If you can’t focus the field iris, check the condenser for a flip-in lens and find the configuration that lets you see the field iris. You may also have to move the field iris into the field of view (step 5) if it is grossly misaligned. 5.Center the field iris with the condenser centering screws. -
To Take Into Consideration the Propriety Of
his was the subject for discussion amongst the seventeen microscopists who met at Edwin Quekett’s house No 50 Wellclose Square, in the Borough of Stepney, East London on 3rd September 1839. It was resolved that such a society be formed Tand a provisional committee be appointed to carry this resolution into effect. The appointed provisional committee of seven were to be responsible for the formation of our society, they held meetings at their homes and drew up a set of rules. They adopted the name ‘Microscopical Society of London’ and arranged a public meeting on the 20th December 1839 at the rooms of the Horticultural Society, 21 Regent Street. Where a Nathaniel Bagshaw Ward © National Portrait Gallery, London President, Treasurer and Secretary were elected, the provisional committee also selected the size of almost airtight containers. Together with George 3 x 1 inch as a standard for glass slides. Loddiges, he saw the potential benefit of protection from sea air damage allowing the transport of plants Each of the members of the provisional committee between continents. This Ward published in 1834 had their own background which we have briefly and eventually his cases enabled the introduction described on the following pages, as you will see of the tea plant to Assam from China and rubber they are a diverse range of professionals. plants to Malaysia from South America. His glass plant cases allowed the growth of orchids and ferns in the Victorian home and in 1842 he wrote a book on the subject. However glass was subject to a tax making cases expensive so Ward lobbied successfully for its repeal in 1845. -
Near-Field Fourier Ptychography: Super- Resolution Phase Retrieval Via Speckle Illumination
Near-field Fourier ptychography: super- resolution phase retrieval via speckle illumination HE ZHANG,1,4,6 SHAOWEI JIANG,1,6 JUN LIAO,1 JUNJING DENG,3 JIAN LIU,4 YONGBING ZHANG,5 AND GUOAN ZHENG1,2,* 1Biomedical Engineering, University of Connecticut, Storrs, CT, 06269, USA 2Electrical and Computer Engineering, University of Connecticut, Storrs, CT, 06269, USA 3Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439, USA. 4Ultra-Precision Optoelectronic Instrument Engineering Center, Harbin Institute of Technology, Harbin 150001, China 5Shenzhen Key Lab of Broadband Network and Multimedia, Graduate School at Shenzhen, Tsinghua University, Shenzhen, 518055, China 6These authors contributed equally to this work *[email protected] Abstract: Achieving high spatial resolution is the goal of many imaging systems. Designing a high-resolution lens with diffraction-limited performance over a large field of view remains a difficult task in imaging system design. On the other hand, creating a complex speckle pattern with wavelength-limited spatial features is effortless and can be implemented via a simple random diffuser. With this observation and inspired by the concept of near-field ptychography, we report a new imaging modality, termed near-field Fourier ptychography, for tackling high- resolution imaging challenges in both microscopic and macroscopic imaging settings. The meaning of ‘near-field’ is referred to placing the object at a short defocus distance with a large Fresnel number. In our implementations, we project a speckle pattern with fine spatial features on the object instead of directly resolving the spatial features via a high-resolution lens. We then translate the object (or speckle) to different positions and acquire the corresponding images using a low-resolution lens. -
The Scientific Legacy of Antoni Van Leeuwenhoek
196 Chapter 12 Chapter 12 The Scientific Legacy of Antoni Van Leeuwenhoek This final chapter discusses some of the developments in science on which Antoni van Leeuwenhoek left his mark from his death to the beginning of the 21st century. It will review the influence of his work and listen for the echoes of his name almost three hundred years after his death. Figure 12.1 Nineteenth-century microscope by George Adams with eyepiece, objective, various attachments and a mirror to illuminate the specimen © Koninklijke Brill NV, Leiden, 2016 | doi 10.1163/9789004304307_013 The Scientific Legacy of Antoni Van Leeuwenhoek 197 Microscopy Microscopes have become increasingly complex and more versatile, but much easier to use, since the time of Van Leeuwenhoek. Single-lens microscopes went out of use in the 18th century, when compound microscopes with at least two lenses ‒ an eyepiece and an objective ‒ became the norm. Many innovations came from England. Firstly, the illumination of speci- mens was improved. During Van Leeuwenhoek’s lifetime, John Marshall (1663–1725) had developed a simple illumination system using a mirror attached to the foot of the microscope. John Cuff (1708–1772) used an extra lens, a condenser, in 1744 to concentrate light on the specimen. In 1755, George Adams (1720–1773) developed a microscope with a rotating wheel holding objectives with different powers of magnification. Sliding holders in which a variety of specimens could be mounted at one time can be traced back to the rotating holders on the single-lensed microscopes used by Christiaan Huygens and J. De Pouilly (or Depovilly) in the 1670s, and were developed for use with compound microscopes. -
Simple and Open 4F Koehler Transmitted Illumination System for Low- Cost Microscopic Imaging and Teaching
Simple and open 4f Koehler transmitted illumination system for low- cost microscopic imaging and teaching Jorge Madrid-Wolff1, Manu Forero-Shelton2 1- Department of Biomedical Engineering, Universidad de los Andes, Bogota, Colombia 2- Department of Physics, Universidad de los Andes, Bogota, Colombia [email protected] ORCID: JMW: https://orcid.org/0000-0003-3945-538X MFS: https://orcid.org/0000-0002-7989-0311 Any potential competing interests: NO Funding information: 1) Department of Physics, Universidad de los Andes, Colombia, 2) Colciencias grant 712 “Convocatoria Para Proyectos De Investigación En Ciencias Básicas“ 3) Project termination grant from the Faculty of Sciences, Universidad de los Andes, Colombia. Author contributions: JMW Investigation, Visualization, Writing (Original Draft Preparation) MFS Conceptualization, Funding Acquisition, Methodology, Supervision, Writing(Original Draft Preparation) 1 Title Simple and open 4f Koehler transmitted illumination system for low-cost microscopic imaging and teaching Abstract Koehler transillumination is a powerful imaging method, yet commercial Koehler condensers are difficult to integrate into tabletop systems and make learning the concepts of Koehler illumination difficult. We propose a simple 4f Koehler illumination system that offers advantages with respect to building simplicity, cost and compatibility with tabletop systems, which can be integrated with open source Light Sheet Fluorescence Microscopes (LSFMs). With those applications in mind as well as teaching, we provide -
160-Series-Manual.Pdf
National Optical & Scientific Instruments Inc. 6508 Tri-County Parkway Schertz, Texas 78154 Phone (210) 590-9010 Fax (210) 590-1104 INSTRUCTIONS FOR 160 SERIES COMPOUND BIOLOGICAL MICROSCOPES Copyright © 1/2/01 National Optical & Scientific Instrument Inc. Models 160 (monocular head), 161 (dual head), 162 (binocular head), 163 (trinocular head), all have the same stand and stand features…only the head portion differs…illustrated is Model 162. Knurled diopter ring Sliding interpupillary adjustment, grips located on both left and right Widefield 10x/18 side of diopter scale eyepiece Mark on side of eyepiece tube for indexing diopter reading Interpupillary scale Knurled head locking screw Viewing head of microscope Revolving nosepiece Arm of microscope stand Objective lenses Two knurled locking screws for Specimen holder securing specimen holder to stage (mechanical stage) Stage Abbe condenser locking screw Abbe condenser 1.25 N.A. Iris diaphragm lever Coarse focus knob Filter holder for 32mm filter Control knob for Fine focus knob Abbe condenser Recess for 45mm filter Illuminator condenser Knobs controlling X and Y movement of mechanical stage Light intensity Base control knob Detachable power supply cord Tension adjustment 100v-240v, 50H/60H input 12 volt DC knob Output ON/OFF switch DC Plug Socket 12 Volt DC Switching Power Supply 3 INTRODUCTION Thank you for your purchase of a National microscope. It is a well built, precision instrument carefully checked to assure that it reaches you in good condition. It is designed for ease of operation and years of carefree use. The information in this manual probably far exceeds what you will need to know in order to operate and maintain your microscope. -
Arxiv:2007.14139V2 [Physics.Optics] 1 Apr 2021 Plane and the Detector Plane
Accelerating ptychographic reconstructions using spectral initializations Lorenzo Valzania,1, 2, ∗ Jonathan Dong,1, 3, ∗ and Sylvain Gigan1 1Laboratoire Kastler Brossel, Ecole´ Normale Sup´erieure - Paris Sciences et Lettres (PSL) Research University, Sorbonne Universit´e,Centre National de la Recherche Scientifique (CNRS) UMR 8552, Coll`egede France, 24 rue Lhomond, 75005 Paris, France 2Laboratory for Transport at Nanoscale Interfaces, Empa, Swiss Federal Laboratories for Materials Science and Technology, 129 Uberlandstrasse, Dubendorf 8600, Switzerland 3Laboratoire de Physique de l'Ecole´ Normale Sup´erieure - Paris Sciences et Lettres (PSL) Research University, Sorbonne Universit´e,CNRS, Universit´ede Paris, 24 rue Lhomond, 75005 Paris, France (Dated: April 2, 2021) Ptychography is a promising phase retrieval technique for label-free quantitative phase imaging. Recent advances in phase retrieval algorithms witnessed the development of spectral methods, in order to accelerate gradient descent algorithms. Using spectral initializations on experimental data, for the first time we report three times faster ptychographic reconstructions than with a standard gradient descent algorithm and improved resilience to noise. Coming at no additional computational cost compared to gradient-descent-based algorithms, spectral methods have the potential to be implemented in large-scale iterative ptychographic algorithms. I. INTRODUCTION and the acquisition parameters. The quest for improved reconstruction performance culminated in the derivation Ptychography is a computational imaging technique of optimal spectral methods for the random measure- that enables label-free, quantitative phase imaging [1]. ments setting [9, 10]. Due to these breakthroughs, spec- It is based on a simple principle: scan a probe across tral methods for phase retrieval are rather well under- a sample, collect the corresponding intensity diffraction stood theoretically. -
Sub-Diffraction Imaging Using Fourier Ptychography and Structured Sparsity
SUB-DIFFRACTION IMAGING USING FOURIER PTYCHOGRAPHY AND STRUCTURED SPARSITY Gauri Jagatap, Zhengyu Chen, Chinmay Hegde, Namrata Vaswani Electrical and Computer Engineering, Iowa State University, Ames, IA 50011 ABSTRACT fewer samples suffice. The assumption of sparsity is natural in sev- eral applications in imaging systems, such as sub-diffraction imag- We consider the problem of super-resolution for sub-diffraction ing, X-ray crystallography, bio-imaging and astronomical imaging imaging. We adapt conventional Fourier ptychographic approaches, [12, 13, 14]; in these applications, the object to be imaged is often for the case where the images to be acquired have an underlying modeled as sparse in the canonical (or wavelet) basis. structured sparsity. We propose some subsampling strategies which Moving beyond sparsity, several algorithms that leverage more can be easily adapted to existing ptychographic setups. We then refined structured sparsity modeling assumptions (such as block use a novel technique called CoPRAM with some modifications, sparsity) achieve considerably improved sample-complexity for re- to recover sparse (and block sparse) images from subsampled pty- constructing images from compressive measurements [15, 16, 17]. chographic measurements. We demonstrate experimentally that this However, analogous algorithms in the context of phase retrieval are algorithm performs better than existing phase retrieval techniques, in not well-studied. In recent previous work [18], we have developed terms of quality of reconstruction, using fewer number of samples. a theoretically-sound algorithmic approach for phase retrieval that Index Terms— Phase retrieval, ptychography, sparsity, non- integrate sparsity (as well as structured sparsity) modeling assump- convex algorithms tions within the reconstruction process. However, that work also assumes certain stringent probabilistic assumptions on the measure- 1. -
How to Perform DIC Microscopy Introduction Principle
How To Perform DIC Microscopy Technical Support Note #T-3 Date Created: 12/5/03 Category:Techniques Date Modified: 00/00/00 Keywords: DIC, Differential Interference Contrast, Axiovert 200M Introduction Many biological specimens cannot be effectively visualized using ordinary bright-field microscopy because their images produce very little contrast, rendering them essentially invisible. Differential Interference Contrast (DIC) microscopy is an excellent method for generating contrast by converting specimen optical path gradients into amplitude differences that can be visualized by the human eye. Images produced using DIC optics have a distinct relief-like, shadow-cast appearance giving an illusion of three dimensionality. This tutorial will cover the optical components used in DIC microscopy, the basic principles governing how it works, and the proper alignment of DIC optical components for the Zeiss Axiovert 200M research microscope. Principle DIC microscopy utilizes a system of dual-beam interference optics that transforms local gradients in optical path length in a specimen into regions of contrast in an image. Optical path length is a product of the refractive index and thickness between two points on an optical path and is directly related to the transit time and number of cycles of vibration exhibited by a wave of light traveling between two points. The basic principle of DIC is illustrated in figure 4. Waves of plane polarized light are split into two beams by the condenser prism. These pairs of beams emerge from the condenser lens traveling in parallel to one another and pass through the specimen. The spacing between these paired beams of light varies depending on the prism used, but generally ranges between 0.2-2µm. -
Setting up Kohler Illumination on the Nikon TE2000U Inverted Microscope
Setting up Kohler Illumination on the Nikon TE2000U Inverted Microscope Koehler illumination1 is an illumination technique that provides optimum resolution and contrast when using a bright-field light microscope. It does this by aligning and focussing the illumination light path, and critically setting the iris apertures of the microscope to best match the objective lens. This sheet is only for aligning the condenser on the Nikon inverted microscope. 1) Place a slide on the stage and bring the specimen into focus. If the microscope condenser is way out of alignment, you will have to roughly adjust the condenser height to give adequate illumination to focus on the specimen. There are two white tape indicators on the left side of the condenser that will be nearly aligned when the condenser is at the correct height. 2) Stop down the Field Diaphragm Iris so that you can see it in the field of view. This is the vertical diaphragm [black] slider at the top right hand of the condenser [at the highest point of the microscope]. You slide the lever downwards to close [stop down] the iris, and up again to re-open the iris. 3) Focus the Condenser by adjusting its height. Turn the focus knob on the left or right side of the condenser until the shadow edge of the Field Diaphragm Iris is in sharp focus when viewed down the microscope eyepieces. This iris edge may be blurred if your specimen is very thick, so get it as sharp in focus as possible. 4) Open the Field Diaphragm Iris to near the edge of the field of view and centre it to the middle, using the condenser centering controls. -
This Is a Repository Copy of Ptychography. White Rose
This is a repository copy of Ptychography. White Rose Research Online URL for this paper: http://eprints.whiterose.ac.uk/127795/ Version: Accepted Version Book Section: Rodenburg, J.M. orcid.org/0000-0002-1059-8179 and Maiden, A.M. (2019) Ptychography. In: Hawkes, P.W. and Spence, J.C.H., (eds.) Springer Handbook of Microscopy. Springer Handbooks . Springer . ISBN 9783030000684 https://doi.org/10.1007/978-3-030-00069-1_17 This is a post-peer-review, pre-copyedit version of a chapter published in Hawkes P.W., Spence J.C.H. (eds) Springer Handbook of Microscopy. The final authenticated version is available online at: https://doi.org/10.1007/978-3-030-00069-1_17. Reuse Items deposited in White Rose Research Online are protected by copyright, with all rights reserved unless indicated otherwise. They may be downloaded and/or printed for private study, or other acts as permitted by national copyright laws. The publisher or other rights holders may allow further reproduction and re-use of the full text version. This is indicated by the licence information on the White Rose Research Online record for the item. Takedown If you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing [email protected] including the URL of the record and the reason for the withdrawal request. [email protected] https://eprints.whiterose.ac.uk/ Ptychography John Rodenburg and Andy Maiden Abstract: Ptychography is a computational imaging technique. A detector records an extensive data set consisting of many inference patterns obtained as an object is displaced to various positions relative to an illumination field.