This book will be useful to those who need an induction to the field of fluorescence or a brief package of information with all the basics but also extending towards the latest in science and technology. As the use of fluorescence techniques in biotechnology and life sciences has advanced rapidly, the book gives a sense of reaching the cutting edge of research. Importantly with its authors spanning a range of disciplines it presents the subject matter from these varying perspectives and gives a thorough coverage of applications. In addition to postgraduate training we hope that the book will also support multi-disciplinary research into applications of fluorescence which require a high degree of interaction of expertise in biology, physics, chemistry, bioengineering and medicine. It is also designed to raise awareness of scientific approaches and technologies that may help to resolve industrially and medically relevant problems in areas such as public health, food processing and safety, and environmental monitoring
The book begins with an introductory level explanation of fluorescence where we revisit the key fundamental concepts and explain the terms. Fluorescence labeling forms an integral part of modern fluorescence technologies. This book covers labeling with fluorescent dyes and genetically encoded fluorescent probes that have revolutionized the life sciences as well as the important emerging fluorescent nanoparticles. Fluorescence labeling coupled with digital imaging leads to new opportunities such as scanning cytometry and spectral imaging as well as challenges of "spectral unmixing" where similar fluorophores are separated by specialized software packages. These are covered in subsequent chapters. Finally we address the fundamental challenge of the correlation of laser scanning microscopy with electron microscopy. We further develop the theme of advanced fluorescence techniques. Among these the spectroscopic and microscopic techniques of Fluorescence Resonant Energy Transfer (FRET) and Fluorescence Recovery After Photobleaching (FRAP) and other photobleaching methods feature prominently. The following chapters focus on various fluorescence technologies, such as time-resolved techniques in fluorescence microscopy, fluorescence correlation spectroscopy (FCS), and flow cytometry, all illustrated with specific applications. In the final part we discuss the applications of fluorescence in broadly understood fields of diagnostic imaging, genetics and clinical biochemistry, as well as for immunochemical detection of analytes. The book concludes with more specific applications in the context of specialised topics such as fluorescence analysis of membranes and probing the kinetics of electrogenic ion pumps via voltage-sensitive fluorescent dyes.
The diversity of a modern biotechnology workplace requires the awareness of the approaches of various disciplines, a demand that is often unmet in conventional postgraduate programs. As the various training institutions across the world continue to adjust their programs to suit this demand, it is not well supported by the existing literature. The most significant barrier and one which most difficult to overcome, divides physics and chemistry from biology. These disciplines have, traditionally, been driven by different paradigms, reinforced by poor overlap of undergraduate degree programs. This needs to be addressed at a postgraduate level so that the new generations of specialists are able to communicate across the specialization gap.