FLIM Microscopy in Biology and MedicineISBN: 978-1-4200-7890-9Suitable for a variety of applications in biology and biomedical research, fluorescence lifetime imaging (FLIM) is a powerful new tool for advanced quantitative analysis. Its high contrast and sensitivity allow for the detection of signals at the single molecule level. After an overview of basic principles of fluorescence, microscopy, and FLIM, this book discusses probes, dynamic and video rate FLIM imaging, laser scanning confocal FLIM, spectral lifetime imaging, nonlinear curve-fitting methods, global FLIM data analysis, and new approaches in data analysis. Widely recognized as pioneers in the field, the editors offer a summary chapter on current FLIM applications.
Molecular Imaging: FRET Microscopy and SpectroscopyISBN: 978-0-19-517720-6FRET microscopy and spectroscopy techniques are important tool used for molecular imaging under physiological conditions. Scientists and clinicians use these microscopy techniques to visualize the protein molecule from living specimens at very high spatial and temporal resolution. This book covers the fundamental principles and theory applied to different FRET microscopy and spectroscopy techniques. The critical tricks and the preparation of specimens required for obtaining a decent FRET image and precise quantitation of the signals from living specimens at the nanomolecular level are also explained. Many critical hints are provided to guide the reader to design and select an appropriate FRET pair on their own depending on their biological system. The aspects of protein molecular imaging are covered in this book are basics, theory, techniques and data analysis - includes basics of fluorescence, fluorophores, microscopy, basics of FRET, detectors, preparation of FRET pairs for cells and tissues, different FRET microscopy systems (wide-field, confocal, multi-photon), FRET data analysis, photobleaching FRET, FLIM-FRET microscopy, Spectral imaging FRET, Single molecule FRET, Time and Image correlation spectroscopy for protein studies and related biological and clinical applications.
Methods in Cellular ImagingISBN: 978-0195139365Advances in technology have revolutionized the development of light microscopy techniques in biomedical research, thus improving visualization of the microstructure of cells and tissues under physiological conditions. Fluorescence microscopy methods are non-contact and non-invasive and provide high spatial and temporal resolution that other laboratory techniques cannot. This well-illustrated book targets graduate students and scientists who are new to the state-of-the-art fluorescence microscopy techniques used in biological and clinical imaging. It explains basic concepts and imaging procedures for wide-field, confocal, multiphoton excitation, fluorescence resonance energy transfer (FRET), lifetime imaging (FLIM), spectral imaging, fluorescence recovery after photobleaching (FRAP), optical tweezers, total internal reflection, high spatial resolution atomic force microscopy (AFM), and bioluminescence imaging for gene expression. The usage of these techniques in various biological applications, including calcium, pH, membrane potential, mitochondrial signal, protein-protein interactions under various physiological conditions, and deep tissue imaging, is clearly presented. The authors describe the approaches to selecting epifluorescence microscopy, the detectors, and the image acquisition and processing software for different biological applications. Step-by-step directions on preparing different digital formats for light microscopy images on websites are also provided.