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.

"There are many critical processes involved in keeping the retina functioning properly. Two of these, the visual cycle and the metabolism of the cell, are tied together by their conversion of important molecules from one form to another. In the visual cycle, 11-cis-retinal is regenerated so that it can combine with a rhodopsin molecule and initiate phototransduction. In cellular metabolism, the cell undergoes many steps to generate adenosine triphosphate, the energy unit of the cell. These mechanisms are critical in maintaining a functioning retina, however they have been difficult to directly interrogate in the living eye. A technique which can quantitatively measure these processes could allow researchers and clinicians to examine them in healthy subjects and how they change under conditions of disease. The goal of this work is to develop a technique which will allow us to investigate these measures of retinal function quantitatively and in a repeatable way. Advantageously, molecules which are converted during the visual cycle or cellular metabolism are accessible using adaptive optics aided two-photon fluorescence ophthalmoscopy. Furthermore, I develop a new technique, adaptive optics fluorescence lifetime ophthalmoscopy, which provides a robust and quantitative measure of a key property of retinal fluorescence. Initially, this method was deployed in a new two-photon adaptive optics ophthalmoscope designed for imaging mice. Exogenous fluorophores with known fluorescence lifetimes were used to validate the initial measurements, before using the new technique to establish baseline measurements for a sensor of cellular metabolism in the mouse eye. Following successful implementation in the mouse, the fluorescence lifetime method was translated to a system dedicated to imaging the macaque retina. By measuring the fluorescence lifetime of endogenous fluorescence originating in the photoreceptors, I found that rods and cones exhibit different fluorescence lifetimes. Further development of this technology may advance research in widespread areas including fluorophore identification in the retina, mechanisms of retinal metabolism, and as a clinical diagnostic."--Pages xiii-xiv.

This open access book provides a comprehensive overview of the application of the newest laser and microscope/ophthalmoscope technology in the field of high resolution imaging in microscopy and ophthalmology. Starting by describing High-Resolution 3D Light Microscopy with STED and RESOLFT, the book goes on to cover retinal and anterior segment imaging and image-guided treatment and also discusses the development of adaptive optics in vision science and ophthalmology. Using an interdisciplinary approach, the reader will learn about the latest developments and most up to date technology in the field and how these translate to a medical setting. High Resolution Imaging in Microscopy and Ophthalmology – New Frontiers in Biomedical Optics has been written by leading experts in the field and offers insights on engineering, biology, and medicine, thus being a valuable addition for scientists, engineers, and clinicians with technical and medical interest who would like to understand the equipment, the applications and the medical/biological background. Lastly, this book is dedicated to the memory of Dr. Gerhard Zinser, co-founder of Heidelberg Engineering GmbH, a scientist, a husband, a brother, a colleague, and a friend.