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Our understanding of the fundamental processes that drive biology and medicine is, in large part, based on our ability to visualize biological structures and monitor their transformations over time. Fluorescence imaging is one of the most transformative technologies of modern biomedical imaging as it provides a low cost, high sensitivity method for real-time molecular imaging in vivo. As the scattering and absorption of light through biological tissue impose significant restrictions on imaging penetration depth, acquisition speed, and spatial resolution, the development of novel optical imaging technologies has increasingly shifted toward the use of light of longer wavelengths. Fluorescence imaging in the shortwave infrared (SWIR, 1000 - 2000 nm) spectral region mitigates the negative effects of light attenuation and benefits from a general lack of tissue autofluorescence. As a result, SWIR imaging promises higher contrast, sensitivity, and penetration depths compared to conventional visible and near-infrared (NIR) fluorescence imaging. However, the lack of versatile and functional SWIR emitters has prevented the general adoption of SWIR imaging both in academic and clinical settings. Here, we will present progress toward the synthesis of a new generation of SWIR-emissive materials and discuss their use in enabling biomedical imaging applications. In the first part of this thesis, we will examine the synthesis of SWIR-emissive indium arsenide (InAs) quantum dots (QDs). To address existing challenges in the synthesis of these semiconductor nanocrystals, we will investigate the processes that govern nanoparticle formation and growth. Combining experimental and theoretical methods, we demonstrate that the synthesis of large nanocrystals is hindered by slow growth rates for large particles, as well as the formation and persistence of small cluster intermediates throughout nanocrystal growth. Based on these insights, we design a novel, rational synthesis for large InAs QDs with high brightness across the SWIR spectral region. Second, we will discuss the use of InAs-based QDs in functional SWIR imaging applications in pre-clinical settings. We will present three QD surface functionalizations that enable the non-invasive real-time imaging of hemorrhagic stroke, the quantification of metabolic activity in genetically-engineered animals, and the measurement of hemodynamics in the brain vasculature of mice. In addition, we will present preliminary results for the synthesis of SWIR-emissive QD probes for the molecular targeting of biological entities and for advanced particle tracking applications. Using a QD-based broadband SWIR emitter, we will further investigate the e↵ect of SWIR imaging wavelength on image contrast and tissue penetration depth. While it was previously assumed that reduced scattering of light at longer wavelengths is the primary cause for increased image contrast, our results indicate that for imaging scenarios with strong fluorescent background signals, image contrast and penetration depth correlate closely with the absorptive properties of biological tissue. As a result, deliberate selection of imaging wavelengths at which biological tissue is highly absorptive can help to overcome contrast-limited imaging scenarios. In the last part of this thesis, we will take a closer look at SWIR emitters with the potential for translation into clinical settings. We will demonstrate that the FDA-approved NIR dye indocyanine green (ICG) exhibits an unexpectedly high SWIR brightness that arises from a large absorption cross-section and a vibronic shoulder in its fluorescence spectrum that extends well into the SWIR spectral region. We expand on this finding by showing that ICG outperforms commercial SWIR dyes during in vivo imaging, and additionally by demonstrating a variety of high-contrast and high-speed imaging applications in small animals. These results suggest that ICG enables the direct translation of SWIR imaging into the clinic. In summary, this thesis will paint a comprehensive picture of the current state of SWIR-emissive materials, present the synthesis of novel versatile SWIR probes, and show their application in unprecedented functional SWIR imaging applications.
Fluorescence imaging offers high spatio-temporal resolution, low radiation dosage exposure, and low cost among all the available imaging modalities, for example, magnetic resonance imaging, computerized tomography and positron emission tomography. Imaging probes of high emissivity and photostability are the key to achieving fluorescence imaging with high signal-to-background ratio (SBR). One promising approach to developing highly bright and stable imaging probes is through surface plasmon enhanced fluorescence. In the first part of the thesis, we develop a fluorescent probe with high site-specificity and emission efficiency by exploiting the targeting-specificity of M13 virus and co-assembling plasmonic nanoparticles and visible dye molecules on the viral capsid. Practical factors controlling fluorescence enhancement, such as nanoparticle size and dye-to-nanoparticle distance, are studied in this project. Lastly, the highly fluorescent probe is applied for in vitro staining of E. coli. The methodology in this work is amendable to developing a wide range of affinity-targeted fluorescent probes using biotemplates. Compared to visible and near infrared spectrum, short-wave infrared (SWIR, 900-1700 nm) spectrum promises high spatial resolution and deep tissue penetration for fluorescence imaging of biological system, owning to low tissue autofluorescence and suppressed tissue scattering at progressively longer wavelengths. In the second part of the thesis, a bright SWIR imaging probe consisting of small SWIR dyes and gold nanorods is developed for in vivo imaging. Fluorescence enhancement is optimized by tuning the dye density on the gold nanorod surface. The SWIR imaging probes are applied for in vivo imaging of ovarian cancer. The effect of targeting modality on intratumor distribution of the imaging probes is studied in two different orthotopic ovarian cancer models. Lastly, we demonstrate that the plasmon enhanced SWIR imaging probe has great potential for fluorescence imaging-guided surgery by showing its capability to detect submillimeter-sized tumors. Apart from enhancing the SWIR down-conversion emission above, surface plasmon enhanced SWIR up-conversion emission is another promising approach to achieving "autofluorescence-free" imaging with minimal tissue scattering. In the third part of the thesis, we use gold nanorods to enhance the up-conversion emission of small SWIR dyes. The mechanism of surface plasmon enhanced up-conversion emission is studied. The up-conversion fluorescence shows much higher SBR than down-conversion fluorescence in non-scatting biological solution and scatting medium. Lastly, we demonstrate in vivo imaging for the first-time using SWIR up-conversion fluorescence with exceptional image contrast.
Discover how metal-enhanced fluorescence is changing traditional concepts of fluorescence This book collects and analyzes all the current trends, opinions, and emerging hot topics in the field of metal-enhanced fluorescence (MEF). Readers learn how this emerging technology enhances the utility of current fluorescence-based approaches. For example, MEF can be used to better detect and track specific molecules that may be present in very low quantities in either clinical samples or biological systems. Author Chris Geddes, a noted pioneer in the field, not only explains the fundamentals of metal-enhanced fluorescence, but also the significance of all the most recent findings and models in the field. Metal-enhanced fluorescence refers to the use of metal colloids and nanoscale metallic particles in fluorescence systems. It offers researchers the opportunity to modify the basic properties of fluorophores in both near- and far-field fluorescence formats. Benefits of metal-enhanced fluorescence compared to traditional fluorescence include: Increased efficiency of fluorescence emission Increased detection sensitivity Protect against fluorophore photobleaching Applicability to almost any molecule, including both intrinsic and extrinsic chromophores Following a discussion of the principles and fundamentals, the author examines the process and applications of metal-enhanced fluorescence. Throughout the book, references lead to the primary literature, facilitating in-depth investigations into particular topics. Guiding readers from the basics to state-of-the-technology applications, this book is recommended for all chemists, physicists, and biomedical engineers working in the field of fluorescence.
The use of light for probing and imaging biomedical media is promising for the development of safe, noninvasive, and inexpensive clinical imaging modalities with diagnostic ability. The advent of ultrafast lasers has enabled applications of nonlinear optical processes, which allow deeper imaging in biological tissues with higher spatial resolution. This book provides an overview of emerging novel optical imaging techniques, Gaussian beam optics, light scattering, nonlinear optics, and nonlinear optical tomography of tissues and cells. It consists of pioneering works that employ different linear and nonlinear optical imaging techniques for deep tissue imaging, including the new applications of single- and multiphoton excitation fluorescence, Raman scattering, resonance Raman spectroscopy, second harmonic generation, stimulated Raman scattering gain and loss, coherent anti-Stokes Raman spectroscopy, and near-infrared and mid-infrared supercontinuum spectroscopy. The book is a comprehensive reference of emerging deep tissue imaging techniques for researchers and students working in various disciplines.
This book focuses on the latest fluorescent materials for cell imaging. Cell imaging is a widely used basic technique that helps scientists gain a better understanding of biological functions through studies of cellular structure and dynamics. In the past decades, the development of a variety of new fluorescent materials has significantly extended the applications of cellular imaging techniques. This book presents recently developed fluorescent materials, including semiconductor quantum dots, carbon dots, silicon nanoparticles, metal nanoclusters, upconversion nanoparticles, conjugated polymers/polymer dots, aggregation-induced emission (AIE) probes, and coordination compounds, used for various cellular imaging purposes. It will appeal to cell biologists and other researchers in academia, industry and clinical settings who are interested in the technical development and advanced applications of fluorescence imaging in cells, tissues and organisms to explore the mechanisms of biological functions and diseases.
In the last decade, bioimaging and therapy based on near-infrared (NIR) nanomaterials have played an important role in biotechnology due to their intrinsic advantages when compared with the traditional imaging probe and medicine. NIR nanomaterials allow deeper penetration depth, low detection threshold concentration and better targeted performance. This book systematically summarises the recent progress in the fabrication and application of NIR nanomaterials for biomedical imaging and therapy, and discusses the advantages, challenges and opportunities available. Near-infrared Nanomaterials contains achapter highlighting the outlook of these materials, detailing novel ideas for the further application of NIR nanomaterials in bioimaging and medicine. Written by leading experts working in the field, this title will have broad appeal to those working in chemistry, materials science, nanotechnology, biology, bioengineering, biomedical science and biophysics.
Functional Fluorescent Materials: Applications in Sensing, Bioimaging, and Optoelectronics explains functional molecular probes (organic/inorganic materials, polymers, nanomaterials), with a focus on those that represent spectroscopic properties with detection of different analytes and specific roles in molecular recognition and their applications. It broadly covers molecular recognition to applications of fluorescence reporters, starting from optoelectronic properties of materials, detection of heavy metals, through biological macromolecules, and further to a living cell, tissue imaging, and theranostics. Features: • Covers different aspects of fluorescence spectroscopy ranging from chemical, physical, and biological aspects along with optoelectronic properties, mechanisms, and applications. • Describes all types of chemical and functionalized fluorescent nanomaterials. • Provides additional information on different kinds of fluorescence reporters. • Explains the concept of fluorescence spectroscopy and its role in human health care. • Discusses changes in static and dynamic properties of fluorescent probes and molecular recognitions. This book is aimed at graduate students and researchers in materials, chemical engineering, and engineering physics.
Nanotechnology for Biomedical Imaging and Diagnostics: From Nanoparticle Design to Clinical Applications reflects upon the increasing role of nanomaterials in biological and medical imaging, presenting a thorough description of current research as well as future directions. With contributions from experts in nanotechnology and imaging from academia, industry, and healthcare, this book provides a comprehensive coverage of the field, ranging from the architectural design of nanomaterials to their broad imaging applications in medicine. Grouped into three sections, the book: Elucidates all major aspects of nanotechnology and bioimaging Provides comprehensive coverage of the field, ranging from the architectural design of nanomaterials to their broad imaging applications in medicine Written by well-recognized experts in academia, industry, and healthcare, will be an excellence source of reference With a multidisciplinary approach and a balance of research and diagnostic topics, this book will appeal to students, scientiests, and healthcare professionals alike
This third edition of a classic text in biological microscopy includes detailed descriptions and in-depth comparisons of parts of the microscope itself, digital aspects of data acquisition and properties of fluorescent dyes, the techniques of 3D specimen preparation and the fundamental limitations, and practical complexities of quantitative confocal fluorescence imaging. Coverage includes practical multiphoton, photodamage and phototoxicity, 3D FRET, 3D microscopy correlated with micro-MNR, CARS, second and third harmonic signals, ion imaging in 3D, scanning RAMAN, plant specimens, practical 3D microscopy and correlated optical tomography.
Written by the prominent art authentication and forgery detection expert David Rudd Cycleback, this small book is a primer on ultraviolet, infrared and visible light in forensic science, art and collectible examination, commerce and daily life. Topics include infrared examination of paintings, currency and license counterfeit detection, invisible ink writing, crime scene investigation, identification of alterations and restoration, and the light techniques used in forgery detection of trading cards, posters, historical documents, art glass and other collectibles. Written for the amateur scientist, junior detective and art and memorabilia collector and dealer.