Abstract
Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM) enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT)] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples.
Highlights
Drug and chemical absorption and disposition in vivo have commonly been characterised indirectly with analytical instrumentation using drug quantitation following tissue sampling
Disadvantages which may limit clinical uses of fluorescence microscopy are the relatively small number of fluorescent stains approved for use in humans and the requirement for additional specialised equipment compared with reflectance imaging
In addition to 5ALA, which is a prodrug of protoporphyrin IX (PPIX), a range of photosensitizers are in clinical use or development and are potentially useful in applications with fluorescence imaging
Summary
Drug and chemical absorption and disposition in vivo have commonly been characterised indirectly with analytical instrumentation using drug quantitation following tissue sampling. Disadvantages which may limit clinical uses of fluorescence microscopy are the relatively small number of fluorescent stains approved for use in humans and the requirement for additional specialised equipment compared with reflectance imaging. The fluorescent dyes in widespread clinical use as injected imaging agents are fluorescein, methylene blue, indocyanine green and the prodrug5aminolevulinate (5ALA) [2,3]. In addition to 5ALA, which is a prodrug of protoporphyrin IX (PPIX), a range of photosensitizers are in clinical use or development and are potentially useful in applications with fluorescence imaging. Recent clinical applications with intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT)] and endomicroscopy. We conclude with a discussion of some existing clinical applications with intravital imaging using endogenous fluorescence or fluorescein stain
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