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Engineering New Capabilities Into Optical Microscopes

Engineering New Capabilities Into Optical Microscopes PDF Author: Matthew D. Lew
Publisher:
ISBN:
Category :
Languages : en
Pages :

Book Description
The labeling of specific biological structures with single fluorescent molecules has ushered in a new era of imaging technology: super-resolution optical microscopy with resolution far beyond the diffraction limit down to some tens of nm. With the features of these exquisite tools in mind, this Dissertation discusses optical strategies for measuring the three-dimensional (3D) position and orientation of single molecules with nanoscale precision and several super-resolution imaging studies of structures in living cells. The concepts of single-molecule imaging, super-resolution microscopy, the engineering of optical point spread functions (PSFs), and quantitative analysis of single-molecule fluorescence images are introduced. The various computational methods and experimental apparatuses developed during the course of my graduate work are also discussed. Next, a new engineered point spread function, called the Corkscrew PSF, is shown for 3D imaging of point-like emitters. This PSF has been demonstrated to measure the location of nanoscale objects with 2-6 nm precision in 3D throughout a 3.2-micrometer depth range. Characterization and application of the Double-Helix (DH) PSF for super-resolution imaging of structures within mammalian and bacterial cells is discussed. The DH-PSF enables 3D single-molecule imaging within living cells with precisions of tens of nanometers throughout a ~2-micrometer depth range. Finally, the impact of single-molecule emission patterns and molecular orientation on optical imaging is treated, with particular emphasis on multiple strategies for improving the accuracy of super-resolution imaging. The DH microscope is shown to be well-suited for accurately and simultaneously measuring the 3D position and orientation of single molecules.

Engineering New Capabilities Into Optical Microscopes

Engineering New Capabilities Into Optical Microscopes PDF Author: Matthew D. Lew
Publisher:
ISBN:
Category :
Languages : en
Pages :

Book Description
The labeling of specific biological structures with single fluorescent molecules has ushered in a new era of imaging technology: super-resolution optical microscopy with resolution far beyond the diffraction limit down to some tens of nm. With the features of these exquisite tools in mind, this Dissertation discusses optical strategies for measuring the three-dimensional (3D) position and orientation of single molecules with nanoscale precision and several super-resolution imaging studies of structures in living cells. The concepts of single-molecule imaging, super-resolution microscopy, the engineering of optical point spread functions (PSFs), and quantitative analysis of single-molecule fluorescence images are introduced. The various computational methods and experimental apparatuses developed during the course of my graduate work are also discussed. Next, a new engineered point spread function, called the Corkscrew PSF, is shown for 3D imaging of point-like emitters. This PSF has been demonstrated to measure the location of nanoscale objects with 2-6 nm precision in 3D throughout a 3.2-micrometer depth range. Characterization and application of the Double-Helix (DH) PSF for super-resolution imaging of structures within mammalian and bacterial cells is discussed. The DH-PSF enables 3D single-molecule imaging within living cells with precisions of tens of nanometers throughout a ~2-micrometer depth range. Finally, the impact of single-molecule emission patterns and molecular orientation on optical imaging is treated, with particular emphasis on multiple strategies for improving the accuracy of super-resolution imaging. The DH microscope is shown to be well-suited for accurately and simultaneously measuring the 3D position and orientation of single molecules.

Harnessing Light

Harnessing Light PDF Author: National Research Council
Publisher: National Academies Press
ISBN: 0309059917
Category : Technology & Engineering
Languages : en
Pages : 358

Book Description
Optical science and engineering affect almost every aspect of our lives. Millions of miles of optical fiber carry voice and data signals around the world. Lasers are used in surgery of the retina, kidneys, and heart. New high-efficiency light sources promise dramatic reductions in electricity consumption. Night-vision equipment and satellite surveillance are changing how wars are fought. Industry uses optical methods in everything from the production of computer chips to the construction of tunnels. Harnessing Light surveys this multitude of applications, as well as the status of the optics industry and of research and education in optics, and identifies actions that could enhance the field's contributions to society and facilitate its continued technical development.

Precision Engineering

Precision Engineering PDF Author: K. Narayanasamy
Publisher: Alpha Science Int'l Ltd.
ISBN: 9788173193514
Category : Computers
Languages : en
Pages : 548

Book Description
Micro-electronics, micro-optics and micro-mechanical components form an integral part of advanced engineered products coming under the broad area of precision engineering. This book covers theme articles and research reports covering the broad area of precision engineering.

Evaluation Engineering

Evaluation Engineering PDF Author:
Publisher:
ISBN:
Category : Electronic apparatus and appliances
Languages : en
Pages : 670

Book Description


Superresolution Optical Microscopy

Superresolution Optical Microscopy PDF Author: Barry R. Masters
Publisher: Springer Nature
ISBN: 3030216918
Category : Science
Languages : en
Pages : 415

Book Description
This book presents a comprehensive and coherent summary of techniques for enhancing the resolution and image contrast provided by far-field optical microscopes. It takes a critical look at the body of knowledge that comprises optical microscopy, compares and contrasts the various instruments, provides a clear discussion of the physical principles that underpin these techniques, and describes advances in science and medicine for which superresolution microscopes are required and are making major contributions. The text fills significant gaps that exist in other works on superresolution imaging, firstly by placing a new emphasis on the specimen, a critical component of the microscope setup, giving equal importance to the enhancement of both resolution and contrast. Secondly, it covers several topics not typically discussed in depth, such as Bessel and Airy beams, the physics of the spiral phase plate, vortex beams and singular optics, photoactivated localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM), and light-sheet fluorescence microscopy (LSFM). Several variants of these techniques are critically discussed. Noise, optical aberrations, specimen damage, and artifacts in microscopy are also covered. The importance of validation of superresolution images with electron microscopy is stressed. Additionally, the book includes translations and discussion of seminal papers by Abbe and Helmholtz that proved to be pedagogically relevant as well as historically significant. This book is written for students, researchers, and engineers in the life sciences, medicine, biological engineering, and materials science who plan to work with or already are working with superresolution light microscopes. The volume can serve as a reference for these areas while a selected set of individual chapters can be used as a textbook for a one-semester undergraduate or first-year graduate course on superresolution microscopy. Moreover, the text provides a captivating account of curiosity, skepticism, risk-taking, innovation, and creativity in science and technology. Good scientific practice is emphasized throughout, and the author’s lecture slides on responsible conduct of research are included as an online resource which will be of interest to students, course instructors, and scientists alike.

Engineering for Surgery

Engineering for Surgery PDF Author: Nicola Pio Belfiore
Publisher: MDPI
ISBN: 303936605X
Category : Technology & Engineering
Languages : en
Pages : 204

Book Description
This Special Issue offers a little taste of the immense variety of an intrinsically vast and interdisciplinary subject, namely, the applications of engineering to surgery. Some of these have been successfully applied or are still under development, while we have been offered a preview of others thanks to the fantasy of creative science fiction writers. This Special Issue aims to stimulate the interest of engineers and surgeons, who will benefit from mutual advantages gained from their cooperation.

US Black Engineer & IT

US Black Engineer & IT PDF Author:
Publisher:
ISBN:
Category :
Languages : en
Pages : 66

Book Description


Fundamentals of Light Microscopy and Electronic Imaging

Fundamentals of Light Microscopy and Electronic Imaging PDF Author: Douglas B. Murphy
Publisher: John Wiley & Sons
ISBN: 1118382935
Category : Medical
Languages : en
Pages : 564

Book Description
Fundamentals of Light Microscopy and Electronic Imaging, Second Edition provides a coherent introduction to the principles and applications of the integrated optical microscope system, covering both theoretical and practical considerations. It expands and updates discussions of multi-spectral imaging, intensified digital cameras, signal colocalization, and uses of objectives, and offers guidance in the selection of microscopes and electronic cameras, as well as appropriate auxiliary optical systems and fluorescent tags. The book is divided into three sections covering optical principles in diffraction and image formation, basic modes of light microscopy, and components of modern electronic imaging systems and image processing operations. Each chapter introduces relevant theory, followed by descriptions of instrument alignment and image interpretation. This revision includes new chapters on live cell imaging, measurement of protein dynamics, deconvolution microscopy, and interference microscopy. PowerPoint slides of the figures as well as other supplementary materials for instructors are available at a companion website: www.wiley.com/go/murphy/lightmicroscopy

Harnessing Light

Harnessing Light PDF Author: National Research Council
Publisher: National Academies Press
ISBN: 0309174392
Category : Technology & Engineering
Languages : en
Pages : 357

Book Description
Optical science and engineering affect almost every aspect of our lives. Millions of miles of optical fiber carry voice and data signals around the world. Lasers are used in surgery of the retina, kidneys, and heart. New high-efficiency light sources promise dramatic reductions in electricity consumption. Night-vision equipment and satellite surveillance are changing how wars are fought. Industry uses optical methods in everything from the production of computer chips to the construction of tunnels. Harnessing Light surveys this multitude of applications, as well as the status of the optics industry and of research and education in optics, and identifies actions that could enhance the field's contributions to society and facilitate its continued technical development.

Atomic Force Microscopy, Scanning Nearfield Optical Microscopy and Nanoscratching

Atomic Force Microscopy, Scanning Nearfield Optical Microscopy and Nanoscratching PDF Author: Gerd Kaupp
Publisher: Springer Science & Business Media
ISBN: 3540284729
Category : Technology & Engineering
Languages : en
Pages : 302

Book Description
Making a clear distinction is made between nano- and micro-mechanical testing for physical reasons, this monograph describes the basics and applications of the supermicroscopies AFM and SNOM, and of the nanomechanical testing on rough and technical natural surfaces in the submicron range down to a lateral resolution of a few nm. New or improved instrumentation, new physical laws and unforeseen new applications in all branches of natural sciences (around physics, chemistry, mineralogy, materials science, biology and medicine) and nanotechnology are covered as well as the sources for pitfalls and errors. It outlines the handling of natural and technical samples in relation to those of flat standard samples and emphasizes new special features. Pitfalls and sources of errors are clearly demonstrated as well as their efficient remedy when going from molecularly flat to rough surfaces. The academic or industrial scientist learns how to apply the principles for tackling their scientific or manufacturing tasks that include roughness far away from standard samples.