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Coherent Anti-Stokes Raman Scattering Microscopy for Biomedical Applications

Coherent Anti-Stokes Raman Scattering Microscopy for Biomedical Applications
Author: Huda Yousif
Publisher:
Total Pages:
Release: 2018
Genre:
ISBN:

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Coherent anti-Stokes Raman scattering (CARS) microscopy is considered as a powerful tool for non-invasive chemical imaging of biological samples. CARS microscopy provides an endogenous contrast mechanism that it is sensitive to molecular vibrations. CARS microscopy is recognized as a great imaging system, especially in vivo experiments since it eliminates the need for the contrast agents. In this thesis, CARS microscopy/spectroscopy is built from scratch by employing a single (Ti-Sapphire) laser source generating 65 femtosecond laser pulses centered at 800 nm wavelength. Two closely lying zero dispersion photonic crystal fiber (PCF) is used to generate the supercontinuum for the Stokes beam to generate CARS at 2885 cm-1 to match lipids rich vibrational frequency. XY galvanometers are used for laser raster scanning across the sample. The initial generation of CARS signal was in the forward direction. After guaranteeing a strong CARS signal, images for chemical and biological samples were taken. To achieve a multimodal imaging technique, CARS microscopy imaging system is combined with two- photon excitation fluorescent (TPEF) and second harmonic generation (SHG) imaging techniques, where various information was extracted from the imaged samples. Images with our CARS microscopy show a good resolution and sensitivity. The second part of my work is to reduce the footprint for this setup to make it more suitable for use in clinical applications. For that reason, I integrated a homebuilt endoscope and all fiber femtosecond laser source together to get a fiber based imaging system. Proof of principal for the integrated system is achieved by obtaining a reasonable agreement in accuracy and resolution to those obtained by the endoscope driven by Ti-sapphire laser.


Coherent Raman Scattering Microscopy

Coherent Raman Scattering Microscopy
Author: Ji-Xin Cheng
Publisher: CRC Press
Total Pages: 603
Release: 2016-04-19
Genre: Science
ISBN: 1439867666

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The First Book on CRS MicroscopyCompared to conventional Raman microscopy, coherent Raman scattering (CRS) allows label-free imaging of living cells and tissues at video rate by enhancing the weak Raman signal through nonlinear excitation. Edited by pioneers in the field and with contributions from a distinguished team of experts, Coherent Raman Sc


Stimulated Raman Scattering Microscopy

Stimulated Raman Scattering Microscopy
Author: Ji-Xin Cheng
Publisher: Elsevier
Total Pages: 612
Release: 2021-12-04
Genre: Science
ISBN: 0323903371

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Stimulated Raman Scattering Microscopy: Techniques and Applications describes innovations in instrumentation, data science, chemical probe development, and various applications enabled by a state-of-the-art stimulated Raman scattering (SRS) microscope. Beginning by introducing the history of SRS, this book is composed of seven parts in depth including instrumentation strategies that have pushed the physical limits of SRS microscopy, vibrational probes (which increased the SRS imaging functionality), data science methods, and recent efforts in miniaturization. This rapidly growing field needs a comprehensive resource that brings together the current knowledge on the topic, and this book does just that. Researchers who need to know the requirements for all aspects of the instrumentation as well as the requirements of different imaging applications (such as different types of biological tissue) will benefit enormously from the examples of successful demonstrations of SRS imaging in the book. Led by Editor-in-Chief Ji-Xin Cheng, a pioneer in coherent Raman scattering microscopy, the editorial team has brought together various experts on each aspect of SRS imaging from around the world to provide an authoritative guide to this increasingly important imaging technique. This book is a comprehensive reference for researchers, faculty, postdoctoral researchers, and engineers. Includes every aspect from theoretic reviews of SRS spectroscopy to innovations in instrumentation and current applications of SRS microscopy Provides copious visual elements that illustrate key information, such as SRS images of various biological samples and instrument diagrams and schematics Edited by leading experts of SRS microscopy, with each chapter written by experts in their given topics


Coherent Anti-Stokes Raman Scattering Miniaturized Microscope

Coherent Anti-Stokes Raman Scattering Miniaturized Microscope
Author: Brett Smith
Publisher:
Total Pages:
Release: 2013
Genre: Microscopes
ISBN:

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Microscopy techniques have been developed and refined over multiple decades, but innovation around single photon modalities has slowed. The advancement of the utility of information acquired, and minimum resolution available is seemingly reaching an asymptote. The fusion of light microscopy and well-studied nonlinear processes has broken through this barrier and enabled the collection of vast amounts of additional information beyond the topographical information relayed by traditional microscopes. Through nonlinear imaging modalities, chemical information can also be extracted from tissue. Nonlinear microscopy also can beat the resolution limit caused by diffraction, and offers up three-dimensional capabilities. The power of nonlinear imaging has been demonstrated by countless research groups, solidifying it as a major player in biomedical imaging. The value of a nonlinear imaging system could be enhanced if a reduction in size would permit the insertion into bodily cavities, as has been demonstrated by linear imaging endoscopes. The miniaturization of single photon imaging devices has led to significant advancements in diagnostics and treatment in the medical field. Much more information can be extracted from a patient if the tissue can be imaged in vivo, a capability that traditional, bulky, table top microscopes cannot offer. The development of new technologies in optics has enabled the miniaturization of many critical components of standard microscopes. It is possible to combine nonlinear techniques with these miniaturized elements into a portable, hand held microscope that can be applied to various facets of the biomedical field. The research demonstrated in this thesis is based on the selection, testing and assembly of several miniaturized optical components for use as a nonlinear imaging device. This thesis is the first demonstration of a fibre delivered, microelectromechanical systems mirror with miniaturized optics housed in a portable, hand held package. Specifically, it is designed for coherent anti-Stokes Raman scattering, second harmonic generation, and two-photon excitation fluorescence imaging. Depending on the modality being exploited, different chemical information can be extracted from the sample being imaged. This miniaturized microscope can be applied to diagnostics and treatments of spinal cord diseases and injuries, atherosclerosis research, cancer tumour identification and a plethora of other biomedical applications. The device that will be revealed in the upcoming text is validated by demonstrating all designed-for nonlinear modalities, and later will be used to perform serialized imaging of myelin of a single specimen over time.


Coherent Anti-Stokes Raman Scattering (CARS) Microscopy for Bioimaging

Coherent Anti-Stokes Raman Scattering (CARS) Microscopy for Bioimaging
Author: Jian Lin
Publisher: LAP Lambert Academic Publishing
Total Pages: 120
Release: 2012
Genre:
ISBN: 9783659289132

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Coherent anti-Stokes Raman scattering (CARS) microscopy is a nonlinear Raman imaging technique that has received great attention for biological and biomedical imaging due to its ability of real-time, nonperturbative chemical mapping of live unstained cells and tissue based on molecular vibrations. However, the strong nonresonant background reduces the image contrast and sensitivity of CARS imaging. Its spatial resolution is limited by light diffraction. This research has systematically studied the near-field (NF) effects of nanoparticle sizes, orientations, polarization of excitations on NF-CARS imaging using FDTD method, and has developed a radially polarized near-field tip-enhanced (TE) CARS system for high-resolution vibrational imaging and a unique annular-aperture detection scheme for suppressing the solvent background. The novel TE-CARS technique and the multimodal nonlinear optical microscopy imaging platform developed in this work have great potential to provide new insights into better understanding of morphological, biochemical and biomolecular changes associated with tissue and cell pathologic transformation at the tissue, cellular and molecular levels without labeling.


Coherent Anti-Stokes Raman Scattering Spectroscopy of Single Molecules in Solution

Coherent Anti-Stokes Raman Scattering Spectroscopy of Single Molecules in Solution
Author:
Publisher:
Total Pages:
Release: 2012
Genre:
ISBN:

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During this funding period, we have developed two breakthrough techniques. The first is stimulated Raman scattering microscopy, providing label-free chemical contrast for chemical and biomedical imaging based on vibrational spectroscopy. Spontaneous Raman microscopy provides specific vibrational signatures of chemical bonds, but is often hindered by low sensitivity. We developed a three-dimensional multiphoton vibrational imaging technique based on stimulated Raman scattering (SRS). The sensitivity of SRS imaging is significantly greater than that of spontaneous Raman microscopy, which is achieved by implementing high-frequency (megahertz) phase-sensitive detection. SRS microscopy has a major advantage over previous coherent Raman techniques in that it offers background-free and readily interpretable chemical contrast. We demonstrated a variety of biomedical applications, such as differentiating distributions of omega-3 fatty acids and saturated lipids in living cells, imaging of brain and skin tissues based on intrinsic lipid contrast, and monitoring drug delivery through the epidermis. This technology offers exciting prospect for medical imaging. The second technology we developed is stimulated emission microscopy. Many chromophores, such as haemoglobin and cytochromes, absorb but have undetectable fluorescence because the spontaneous emission is dominated by their fast non-radiative decay. Yet the detection of their absorption is difficult under a microscope. We use stimulated emission, which competes effectively with the nonradiative decay, to make the chromophores detectable, as a new contrast mechanism for optical microscopy. We demonstrate a variety of applications of stimulated emission microscopy, such as visualizing chromoproteins, non-fluorescent variants of the green fluorescent protein, monitoring lacZ gene expression with a chromogenic reporter, mapping transdermal drug distribu- tions without histological sectioning, and label-free microvascular imaging based on endogenous contrast of haemoglobin. For all these applications, sensitivity is orders of magnitude higher than for spontaneous emission or absorption contrast, permitting nonfluorescent reporters for molecular imaging. Although we did not accomplish the original goal of detecting single-molecule by CARS, our quest for high sensitivity of nonlinear optical microscopy paid off in providing the two brand new enabling technologies. Both techniques were greatly benefited from the use of high frequency modulation for microscopy, which led to orders of magnitude increase in sensitivity. Extensive efforts have been made on optics and electronics to accomplish these breakthroughs.


Handbook of Biomedical Nonlinear Optical Microscopy

Handbook of Biomedical Nonlinear Optical Microscopy
Author: Barry R. Masters
Publisher: Oxford University Press
Total Pages: 895
Release: 2008-05-19
Genre: Science
ISBN: 0198036825

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The Handbook of Biomedical Nonlinear Optical Microscopy provides comprehensive treatment of the theories, techniques, and biomedical applications of nonlinear optics and microscopy for cell biologists, life scientists, biomedical engineers, and clinicians. The chapters are separated into basic and advanced sections, and provide both textual and graphical illustrations of all key concepts. The more basic sections are aimed at life scientists without advanced training in physics and mathematics, and tutorials are provided for the more challenging sections. The first part of the Handbook introduces the historical context of nonlinear microscopy. The second part presents the nonlinear optical theory of two- and multiphoton excited fluorescence (TPE, MPE) spectroscopy, second and third harmonic generation (SHG, THG) spectroscopy, and coherent anti-Stokes Raman spectroscopy (CARS). The third part introduces modern microscopic and spectroscopic instrumentation and techniques that are based on nonlinear optics. The fourth part provides key applications of nonlinear microscopy to the biomedical area: neurobiology, immunology, tumor biology, developmental biology, dermatology, and cellular metabolism. There are also chapters on nonlinear molecular probes, cellular damage, and nanoprocessing.


Principles of Nonlinear Optical Spectroscopy

Principles of Nonlinear Optical Spectroscopy
Author: Shaul Mukamel
Publisher: Oxford University Press on Demand
Total Pages: 543
Release: 1999
Genre: Science
ISBN: 9780195132915

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This textbook presents a systematic and unifying viewpoint for a wide class of nonlinear spectroscopic techniques in time domain and frequency domain. It is directed towards active researchers in physics, optics, chemistry, and materials science, as well as graduate students who enter this complex and rapidly developing field. Nonlinear optical interactions of laser fields with matter provide powerful spectroscopic tools for the understanding of microscopic interactions and dynamic processes. One of the major obstacles facing researchers in this field, however, is the flood of experimental techniques and terminologies, which create a serious language barrier. The general microscopic correlation function approach to the nonlinear optical response developed in this book is essential for understanding the relationships among different techniques and a comparison of their information content, the design of new measurements, and for a systematic comparison of the optical response of different systems such as dyes in solutions, atoms and molecules in the gas phase, liquids, molecular aggregates and superlatives, and semiconductor nanostructures. The approach is based on formulating the nonlinear response by representing the state of matter by the density matrix and following its evolution on Liouville space. Current active research areas such as femtosecond time-domain techniques, semi-classical and wave-packet dynamics, pulse shaping, pulse locking, exciton confinement, and the interplay of electronic, nuclear and field coherence are emphasized. The material has been developed from the author's highly successful interdisciplinary course at the University of Rochester attended by science and engineering graduate students.


Vibrational Spectroscopic Imaging for Biomedical Applications

Vibrational Spectroscopic Imaging for Biomedical Applications
Author: Gokulakrishnan Srinivasan
Publisher: McGraw Hill Professional
Total Pages: 381
Release: 2010-06-05
Genre: Technology & Engineering
ISBN: 0071597085

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The latest advances in vibrational spectroscopic biomedical imaging Written by expert spectroscopists, Vibrational Spectroscopic Imaging for Biomedical Applications discusses recent progress in the field in areas such as instrumentation, detector technology, novel modes of data collection, data analysis, and various biomedical applications. This full-color volume covers various IR imaging techniques, including transmission reflection, transflection, and attenuated total reflection (ATR) imaging, and Raman imaging. The efficient use of vibrational spectroscopy in clinical applications is emphasized in this state-of-the-art guide. Coverage includes: Automated breast histopathology using mid-IR spectroscopic imaging Synchrotron-based FTIR spectromicroscopy and imaging of single algai cells and cartilage Preparation of tissues and cells for infrared and Raman spectroscopy and imaging Evanescent wave imaging sFTIR, Raman, and surface-enhanced Raman spectroscopic imaging of fungal cells Widefield Raman imaging of cells and tissues Resonance Raman imaging and quantification of carotenoid antioxidants in the human retina and skin Raman microscopy for biomedical applications--efficient diagnosis of tissues, cells, and bacteria The current sate of Raman imaging in clinical application Vibrational spectroscopic imaging of microscopic stress patterns in biomedical materials Tissue imaging with coherent anti-Stokes Raman scattering microscopy