Fundamentals of Fluorescence Microscopy: Exploring Life with Light

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This book starts at an introductory level and leads reader to the most advanced topics in fluorescence imaging and super-resolution techniques that have enabled new developments such as nanobioimaging, multiphoton microscopy, nanometrology and nanosensors.
The interdisciplinary subject of fluorescence microscopy and imaging requires complete knowledge of imaging optics and molecular physics. So, this book approaches the subject by introducing optical imaging concepts before going in more depth about advanced imaging systems and their applications. Additionally, molecular orbital theory is the important basis to present molecular physics and gain a complete understanding of light-matter interaction at the geometrical focus. The two disciplines have some overlap since light controls the molecular states of molecules and conversely, molecular states control the emitted light. These two mechanisms together determine essential imaging factors such as, molecular cross-section, Stoke shift, emission and absorption spectra, quantum yield, signal-to-noise ratio, Forster resonance energy transfer (FRET), fluorescence recovery after photobleaching (FRAP) and fluorescence lifetime. These factors form the basis of many fluorescence based devices.
The book is organized into two parts. The first part deals with basics of imaging optics and its applications. The advanced part takes care of several imaging techniques and related instrumentation that are developed in the last decade pointing towards far-field diffraction unlimited imaging.

Author(s): Partha Pratim Mondal, Alberto Diaspro (auth.)
Edition: 1
Publisher: Springer Netherlands
Year: 2014

Language: English
Pages: 218
Tags: Spectroscopy and Microscopy; Biological Microscopy; Biophysics and Biological Physics; Optics, Optoelectronics, Plasmonics and Optical Devices; Spectroscopy/Spectrometry

Front Matter....Pages I-XV
Front Matter....Pages 1-1
Ray Optics, Wave Optics and Imaging System Design....Pages 3-31
Basics of Electromagnetic Theory for Fluorescence Microscopy....Pages 33-51
Electric Field Effects in Optical Microscopy Systems....Pages 53-70
Quantum Description of Radiation Field and Optical Microscopy....Pages 71-94
Molecular Physics of Fluorescent Markers....Pages 95-107
Front Matter....Pages 109-109
Basics of Fluorescence and Photophysics....Pages 111-134
General Fluorescence Imaging Techniques....Pages 135-147
Multiphoton Fluorescence Microscopy....Pages 149-159
Super-resolution Fluorescence Microscopy....Pages 161-187
Image Reconstruction for Fluorescence Microscopy....Pages 189-202
Future Prespective of Fluorescence Microscopy....Pages 203-207
Back Matter....Pages 209-218