With the advent of Fourier-domain techniques, optical coherence tomography (OCT) has advanced from high-resolution ‘point’ imaging over small fields-of-view to comprehensive microscopic imaging over three-dimensional volumes that are comparable to the dimensions of luminal internal organs. This advance has required the development of new lasers, improved spectrometers, minimally invasive catheters and endoscopes, and novel optical and signal processing strategies. In recent cardiovascular, ophthalmic, and gastrointestinal clinical studies, the capabilities of Fourier-domain OCT have enabled a new paradigm for diagnostic screening of large tissue areas, which addresses the shortcomings of existing technologies and focal biopsy.