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Author: Aaron Mok Publisher: ISBN: Category : Languages : en Pages : 0
Book Description
Imaging large populations of neurons across multiple brain regions at substantial depth are necessary to understand animal behaviors. However, this is challenging for optical imaging due to: (a) optical aberrations, scattering, and absorption in biological tissue, and (b) the number of neurons that can be recorded is limited by the "photon budget"-under a maximum allowable average power on the biological sample, only a limited number of neurons can be imaged. This limits both imaging depth and imaging width in multiphoton imaging.In this thesis, we first report a simple and versatile tissue spectrometer to measure the optical scattering and absorption in biological samples in terms of ballistic and total transmittance at a wavelength from 450 nm to 1630 nm. The measurement results are important to determine the optimal excitation wave length and fluorophores for deep imaging. With measurements showing that fly head cuticle has high transmission at wavelengths of > 900 nm, we were able to develop a multiphoton imaging method to capture neural structure and activity in behaving flies through the intact cuticles. This through-cuticle imaging method extends the time limits of in-vivo imaging in flies and opens new avenues for imaging the neuronal structure and activity of an intact fly brain. Challenges posed by the photon budget for deep and wide imaging in multiphoton imaging can be partially mitigated by careful optimization of the excitation wavelength, excitation focus profile, and allocation of laser power. We carefully compared the excitation efficiency of Gaussian focus, variations of Gaussian focus, and Bessel focus. We found that for neuronal activity imaging in three-photon microscopy, using multiple foci on the same neuron with an enlarged Gaussian focus of _ 5 - 10 μm axial extension provides the best spike detection accuracy. Finally, we developed a synergistic combination of using multiple foci and adaptive excitation to reduce the average power required on the sample through the intelligent allocation of laser power only on the regions of interest. These optimizations lead to a more efficient imaging scheme in three-photon excitation. It enabled us to improve the efficiency, scanning speed, and field-of-view of three-photon microscopy without exceeding the allowable power limit. We built a large field-of-view multiphoton microscope for two- and three-photon imaging and demonstrated three-photon imaging with a field-of-view of _ 3.5 mm in diameter and > 1 mm in depth across various mouse brain regions and across the entire width of the zebrafish brain.
Author: Aaron Mok Publisher: ISBN: Category : Languages : en Pages : 0
Book Description
Imaging large populations of neurons across multiple brain regions at substantial depth are necessary to understand animal behaviors. However, this is challenging for optical imaging due to: (a) optical aberrations, scattering, and absorption in biological tissue, and (b) the number of neurons that can be recorded is limited by the "photon budget"-under a maximum allowable average power on the biological sample, only a limited number of neurons can be imaged. This limits both imaging depth and imaging width in multiphoton imaging.In this thesis, we first report a simple and versatile tissue spectrometer to measure the optical scattering and absorption in biological samples in terms of ballistic and total transmittance at a wavelength from 450 nm to 1630 nm. The measurement results are important to determine the optimal excitation wave length and fluorophores for deep imaging. With measurements showing that fly head cuticle has high transmission at wavelengths of > 900 nm, we were able to develop a multiphoton imaging method to capture neural structure and activity in behaving flies through the intact cuticles. This through-cuticle imaging method extends the time limits of in-vivo imaging in flies and opens new avenues for imaging the neuronal structure and activity of an intact fly brain. Challenges posed by the photon budget for deep and wide imaging in multiphoton imaging can be partially mitigated by careful optimization of the excitation wavelength, excitation focus profile, and allocation of laser power. We carefully compared the excitation efficiency of Gaussian focus, variations of Gaussian focus, and Bessel focus. We found that for neuronal activity imaging in three-photon microscopy, using multiple foci on the same neuron with an enlarged Gaussian focus of _ 5 - 10 μm axial extension provides the best spike detection accuracy. Finally, we developed a synergistic combination of using multiple foci and adaptive excitation to reduce the average power required on the sample through the intelligent allocation of laser power only on the regions of interest. These optimizations lead to a more efficient imaging scheme in three-photon excitation. It enabled us to improve the efficiency, scanning speed, and field-of-view of three-photon microscopy without exceeding the allowable power limit. We built a large field-of-view multiphoton microscope for two- and three-photon imaging and demonstrated three-photon imaging with a field-of-view of _ 3.5 mm in diameter and > 1 mm in depth across various mouse brain regions and across the entire width of the zebrafish brain.
Author: F.S. Pavone Publisher: IOS Press ISBN: 1614994137 Category : Science Languages : en Pages : 219
Book Description
Biophotonics and microscopy are highly inter-related fields in terms of both technological development and biomedical applications. Recent advances in microscopy have been paralleled by new opportunities for biophotonics, including the investigation and manipulation of biological phenomena using light and its application to biomedicine. This book contains papers from the Enrico Fermi International School of Physics on Microscopy Applied to Biophotonics, held in Varenna, Italy, in July 2011. The lectures spanned the basic science of imaging, through advanced microscopy techniques, to the state-of-the-art in biomedical imaging, and were complemented by seminars from world leaders in biophotonics. Subjects covered include: an overview of biophotonics; fundamentals of microscopy and an introduction to nonlinear microscopy; fluorescence; lasers for biophotonics; and an introduction to ultra-microscopy.
Author: Edyth L. Malin Publisher: Springer Science & Business Media ISBN: 1461519136 Category : Science Languages : en Pages : 398
Book Description
Although the art of making cheese can be traced to prehistoric times, it has continued to evolve as modern civilization progressed. The advent of new technologies and instrumentation has brought exponential growth in the understanding of cheese components and their function. Even more recently, the evolution of cheesemaking has accelerated, driven by economic factors such as the establishment of the European Economic Community, the changing diet of developed countries, and the environmental and economic concerns associated with whey disposal. Molecular biology has revolutionized the development of starter and adjunct cultures as well as rennets, and genetics will make it possible to maintain ideal milk components for cheesemaking. The ability to accelerate traditional ripening procedures has altered the production of certain cheeses, and the emphasis on decreasing the intake of dietary fat, especially in the United States, has prompted the development of technology for producing low-fat cheeses with traditional texture and flavor. In assembling a distinguished group of participants for the symposium, "Chemistry of the Structure/Function Relationships in Cheese," we hoped to review the interplay of these trends and forecast the direction of future research. Contributors evaluated the current status of cheesemaking and highlighted the information that will be essential for new developments. They also focused the attention of agricultural and food chemists on the opportunities in cheese research and the potential contributions they might make to the future of cheese, a most valuable food product. We are indebted to Dr. Patrick Fox, Dr. Mark Johnson, Dr. Milos Kalab, Dr.
Author: Francesco S. Pavone Publisher: Taylor & Francis ISBN: 1439849153 Category : Science Languages : en Pages : 465
Book Description
Second-harmonic generation (SHG) microscopy has shown great promise for imaging live cells and tissues, with applications in basic science, medical research, and tissue engineering. Second Harmonic Generation Imaging offers a complete guide to this optical modality, from basic principles, instrumentation, methods, and image analysis to biomedical a
Author: Robert R. Alfano Publisher: William Andrew ISBN: 0323480683 Category : Technology & Engineering Languages : en Pages : 612
Book Description
Neurophotonics and Biomedical Spectroscopy addresses the novel state-of-the-art work in non-invasive optical spectroscopic methods that detect the onset and progression of diseases and other conditions, including pre-malignancy, cancer, Alzheimer's disease, tissue and cell response to therapeutic intervention, unintended injury and laser energy deposition. The book then highlights research in neurophotonics that investigates single and multi-photon excitation optical signatures of normal/diseased nerve tissues and in the brain, providing a better understanding of the underlying biochemical and structural changes of tissues and cells that are responsible for the observed spectroscopic signatures. Topics cover a wide array of well-established UV, visible, NIR and IR optical and spectroscopic techniques and novel approaches to diagnose tissue changes, including: label free in vivo and ex vivo fluorescence spectroscopy, Stoke shift spectroscopy, spectral imaging, Resonance Raman spectroscopy, multiphoton two Photon excitation, and more. - Provides an overview of the spectroscopic properties of tissue and tissue-light interaction, describing techniques to exploit these properties in imaging - Explores the potential and significance of molecule-specific imaging and its capacity to reveal vital new information on nanoscale structures - Offers a concise overview of different spectroscopic methods and their potential benefits for solving diagnostic and therapeutic problems
Author: Ruikang K. Wang Publisher: Taylor & Francis ISBN: 1439895821 Category : Science Languages : en Pages : 726
Book Description
Despite a number of books on biophotonics imaging for medical diagnostics and therapy, the field still lacks a comprehensive imaging book that describes state-of-the-art biophotonics imaging approaches intensively developed in recent years. Addressing this shortfall, Advanced Biophotonics: Tissue Optical Sectioning presents contemporary methods and
Author: Karsten König Publisher: Walter de Gruyter GmbH & Co KG ISBN: 311043007X Category : Science Languages : en Pages : 785
Book Description
This monograph focuses on modern femtosecond laser microscopes for two photon imaging and nanoprocessing, on laser tweezers for cell micromanipulation as well as on fluorescence lifetime imaging (FLIM) in Life Sciences. The book starts with an introduction by Dr. Wolfgang Kaiser, pioneer of nonlinear optics and ends with the chapter on clinical multiphoton tomography, the novel high resolution imaging technique. It includes a foreword by the nonlinear microscopy expert Dr. Colin Sheppard. Contents Part I: Basics Brief history of fluorescence lifetime imaging The long journey to the laser and its use for nonlinear optics Advanced TCSPC-FLIM techniques Ultrafast lasers in biophotonics Part II: Modern nonlinear microscopy of live cells STED microscopy: exploring fluorescence lifetime gradients for super-resolution at reduced illumination intensities Principles and applications of temporal-focusing wide-field two-photon microscopy FLIM-FRET microscopy TCSPC FLIM and PLIM for metabolic imaging and oxygen sensing Laser tweezers are sources of two-photon effects Metabolic shifts in cell proliferation and differentiation Femtosecond laser nanoprocessing Cryomultiphoton imaging Part III: Nonlinear tissue imaging Multiphoton Tomography (MPT) Clinical multimodal CARS imaging In vivo multiphoton microscopy of human skin Two-photon microscopy and fluorescence lifetime imaging of the cornea Multiscale correlative imaging of the brain Revealing interaction of dyes and nanomaterials by multiphoton imaging Multiphoton FLIM in cosmetic clinical research Multiphoton microscopy and fluorescence lifetime imaging for resection guidance in malignant glioma surgery Non-invasive single-photon and multi-photon imaging of stem cells and cancer cells in mouse models Bedside assessment of multiphoton tomography
Author: Bingbing Cheng Publisher: ISBN: Category : Diagnostic imaging Languages : en Pages : 135
Book Description
Fluorescence microscopic imaging in centimeter-deep tissue has been highly sought-after for many years because much interesting in vivo micro-information--such as microcirculation, tumor angiogenesis, and cancer metastasis--may deeply locate in tissue. However, it is very challenging because of strong tissue light scattering. This includes: how to confine the fluorescence emission into a small volume to achieve high spatial resolution; how to increase fluorescence emission efficiency to compensate the signal attenuation caused by small emission volume and tissue scattering/absorption; and how to reduce background fluorescence noise and exclusively differentiate signal photons from background photons to increase signal-to-noise ratio (SNR) and sensitivity. Ultrasonic scattering is two to three orders of magnitude less than light scattering in opaque biological tissue. Therefore, light focusing has been replaced by ultrasonic focusing to achieve high spatial resolution in deep tissue. In addition, high intensity focused ultrasound (HIFU) can noninvasively heat a small region deep within the body (hundreds of microns in lateral). If one can develop a contrast agent whose fluorescence emission is sensitive to this HIFU-induced temperature change and a sensitive imaging system which can detect the ultrasound-controlled photons that have been scattered many times, centimeter-deep tissue fluorescence microscopic imaging can be achievable. This study is focused on developing a fundamentally different imaging technology: ultrasound-switchable fluorescence (USF), including the contrast agent development and the imaging system development. Basically, the USF contrast agent developed in this work is thermosensitive and its fluorescence emission has a switch-like relationship with temperature. Its fluorescence emission can be switched on or off by a focused ultrasound beam generated from a HIFU transducer within its focal volume. Then the diffused USF photons propagate out and are detected by a sensitive USF imaging system. First, the excellent USF imaging contrast agents were developed by using the environment-sensitive fluorophores and thermosensitive polymers. We started investigating environment-sensitive fluorophores from visible light range up to near infrared (NIR) range since only NIR light can penetrate centimeter-deep in opaque biological tissues. Two basic thermosensitive polymers and their co-polymers were used, including: poly (N-isopropylacrylamide) (PNIPAM) and pluronic. Both linear polymer and nanoparticle based USF contrast agents were explored. Second, a sensitive frequency domain (FD) USF imaging system and an effective signal identification algorithm were developed. The lock-in amplifier adopted in the FD-USF imaging system and the correlation algorithm significantly improved the SNR and detection sensitivity. Third, the feasibility of USF imaging in centimeter-deep tissue with high resolution was demonstrated in both tissue-mimicking phantoms and ex vivo biological tissues. Multi-color high-resolution USF imaging in centimeter-deep tissue with high SNR and picomole sensitivity were also achieved. Fourth, the feasibility of in vivo USF imaging was demonstrated in living mice by using different types of USF contrast agents via both intravenous and local injections. In summary, the results provided in this work demonstrated for the first time the feasibility of centimeter-deep tissue fluorescence microscopic imaging with high SNR and picomole sensitivity via USF in tissue-mimicking phantoms, porcine muscle tissues, ex vivo mouse organs (liver and spleen), and in vivo mice. Multiplex USF imaging was also achieved, which is useful to simultaneously image multiple targets and observe their interactions. This work opens the door for future studies of center-deep tissue fluorescence microscopic imaging.
Author: Lingyan Shi Publisher: CRC Press ISBN: 1351797387 Category : Medical Languages : en Pages : 444
Book Description
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.
Author: Aaron Mok
Author: Aaron Mok
Author: Adrià Escobet Montalbán
Author: F.S. Pavone
Author: Edyth L. Malin
Author: Francesco S. Pavone
Author: Robert R. Alfano
Author: Ruikang K. Wang
Author: Karsten König
Author: Bingbing Cheng
Author: Lingyan Shi