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During the past two decades, there has been an increasing appreciation of the significant value that lifetime-based techniques can add to biomedical studies and applications of fluorescence. Bringing together perspectives of different research communities, Fluorescence Lifetime Spectroscopy and Imaging: Principles and Applications in Biomedical Diagnostics explores the remarkable advances in time-resolved fluorescence techniques and their role in a wide range of biological and clinical applications. Broadly accessible, the book captures the state-of-the-art of fluorescence lifetime metrology and imaging and provides current perspectives on their applications to biomedical studies of intact tissues and medical diagnosis. The text introduces these techniques within the wider context of fluorescence spectroscopy and describes basic principles underlying current instrumentation for fluorescence lifetime imaging and metrology (FLIM). It also covers the wide range of methods, including single channel (point) spectroscopy, fluorescence lifetime imaging microscopy, and single- and multi-photon excitation. Edited by pioneers in this field, with contributions from leading experts, the book includes an overview of complementary techniques that help researchers beginning FLIM research. It offers a comprehensive treatment of fundamental principles, instrumentation, analytical methods, and applications. It also provides an overview of the label-free contrast available from lifetime measurements of tissue autofluorescence and the prospects for exploiting this for clinical applications and biomedical research including drug discovery.
This book focuses on the emerging non-invasive imaging technique of Fluorescence Lifetime Imaging Ophthalmoscopy (FLIO). FLIO reveals unique information on retinal diseases, ranging from age-related macular degeneration and vascular diseases to hereditary retinal dystrophies. Fluorescence lifetimes enable the evaluation of disease progression before irreversible structural changes occur. The image acquisition is suitable for diagnostic purposes and follow-up examinations to investigate the natural course of disease, and to monitor the effects of possible therapies. This book fills the gap between available literature and gives state-of-the-art guidance on the principles of the FLIO technique, image acquisition, and data analysis. Written by a team of expert leaders within this field, this book will be relevant for scientists and clinicians with an interest in ophthalmoscopy.
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
This is an interdisciplinary book that presents the applications of novel laser spectroscopy and imaging techniques for the detection of cancers recently developed by some of the world's most renown researchers. The book consists of three parts and a total of 16 chapters. Each chapter is written by leading experts who are actively seeking to develop novel spectroscopic and analytical methods for cancer detection and diagnosis.In Part I, the authors present fundamentals on optics, atoms and molecules, biophysics, cancer and machine learning. These chapters are intended for those who are not experts in the field but wish to learn about fundamentals' aspects of some of the key topics that are addressed in this book. Particular attention has been given to providing key references for those who wish to go further into the fundamental aspects of atoms and molecules, light-matter interaction, optical instrumentation, machine learning and cancer.In Part II, the authors present key applications of various laser spectroscopic methods in cancer diagnosis. They have provided recent progress in cancer diagnostics obtained by combining laser spectroscopy and machine learning for the analysis of the spectra acquired from biomedical tissues and biofluids.In Part III, the authors present chapters that discuss key developments in the applications of various laser imaging techniques for cancer detection.This is one of the few books that addresses cancer detection and diagnosis using laser spectroscopic and imaging tools with an eye on providing the reader the scientific tools, including machine learning ones.
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
This book provides an in-depth description and discussion of different multi-modal diagnostic techniques for cancer detection and treatment using exact optical methods, their comparison, and combination. Coverage includes detailed descriptions of modern state of design for novel methods of optical non-invasive cancer diagnostics; multi-modal methods for earlier cancer diagnostic enhancing the probability of effective cancer treatment; modern clinical trials with novel methods of clinical cancer diagnostics; medical and technical aspects of clinical cancer diagnostics, and long-term monitoring. Biomedical engineers, cancer researchers, and scientists will find the book to be an invaluable resource. Introduces optical imaging strategies; Focuses on multimodal optical diagnostics as a fundamental approach; Discusses novel methods of optical non-invasive cancer diagnostics.
During the past two decades, there has been an increasing appreciation of the significant value that lifetime-based techniques can add to biomedical studies and applications of fluorescence. Bringing together perspectives of different research communities, Fluorescence Lifetime Spectroscopy and Imaging: Principles and Applications in Biomedical Dia
This volume provides resources, ideas, and bench manuals for the study of breast cancer. This book is divided into five sections: methods used in clinical laboratory for diagnosis (Detection of Molecular Markers of Breast Cancer); methods used in both clinical and research laboratories for testing genetic alterations (Genetic Detection of Breast Cancer) and (Isolation of Breast Cancer Cells); methods used to study the behavior of breast cancer cells (In Vitro Experimental Assays for Breast Cancer); and methods used for mimicking human breast cancer in a living organism (In Vivo Experimental Models for Breast Cancer). Breast Cancer: Methods and Protocols also explores several recently developed techniques for the study of breast cancer progression. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and thorough, Breast Cancer: Methods and Protocols, is a valuable handbook for both graduate and advanced undergraduate students of biological sciences, as well as scientists, technicians, and physicians working in the academic, hospital, or pharmaceutical industry aimed at studying or diagnosing breast cancer.
From the Lab to Clinical Settings—Advances in Quantitative, Noninvasive Optical Diagnostics Noninvasive fluorescence imaging techniques, novel fluorescent labels, and natural biomarkers are revolutionizing our knowledge of cellular processes, signaling and metabolic pathways, the underlying mechanisms for health problems, and the identification of new therapeutic targets for drug discoveries. Natural Biomarkers for Cellular Metabolism: Biology, Techniques, and Applications delves into the current state of knowledge on intrinsic fluorescent biomarkers and highlights recent developments in using these biomarkers for the metabolic mapping and clinical diagnosis of healthy and diseased cells and tissues. Autofluorescent Biomarkers for Biomedical Diagnostics The book’s first section introduces the fundamentals of cellular energy metabolism as well as natural biomarkers within the context of their biological functions. The second section outlines the theoretical and technical background of quantitative, noninvasive, autofluorescence microscopy and spectroscopy methods, including experimental design, calibration, pitfalls, and remedies of data acquisition and analysis. The last two sections highlight advances in biomedical and biochemical applications, such as monitoring stem cell differentiation in engineered tissues and diagnosing cancer and ophthalmic diseases quantitatively and noninvasively. Tailored to Interdisciplinary Researchers Covering cell biology, imaging techniques, and clinical diagnostics, this book provides readers with a complete guide to studying cellular/tissue metabolism under healthy, diseased, and environment-induced stress conditions using natural biomarkers. The book is designed for graduate and advanced undergraduate students, biophysics instructors, medical researchers, and those in pharmaceutical R&D.