In vivo confocal imaging: general principles and applications

It is well established that confocal microscopy provides higher resolution images with better rejection of out-of-focus information than conventional light microscopy. The optical sectioning ability of confocal microscopy allows images to be obtained from different depths within a thick tissue specimen, thereby eliminating the need for processing and sectioning procedures. Thus, confocal microscopy has made it possible to view biological tissues under more physiologic conditions than previously possible. The most widespread biological application of confocal microscopy has been in the localization of immunofluorescently labeled proteins in cell culture or within excised blocks of tissue. Because of its noninvasive optical sectioning capability, confocal microscopy is also ideally suited to the study of tissue in intact living animals, although the potential in vivo applications of this paradigm have received less attention. In this paper we trace the development of in vivo confocal microscopy and present examples of current capabilities for both research and clinical use.