ABSTRACT
This chapter presents the most commonly used methods for optical imaging, looking at methods in which the light field is limited in extent, or spatially controlled, and also where novel fluorophores are combined with high-speed sensitive cameras and advanced computing to build up images in effect molecule by molecule. The instruments use high-energy beams of electrons, which have a very short wavelength, and hence the ability to image with nanometer resolution. Total internal reflection microscopy is, in some sense, not a full super-resolution technique as it only provides super resolution in the axial direction. Structured illumination microscopy (SIM) is another widefield, super-resolution technique, though in comparison to total internal reflection microscopy, SIM produces sub-diffraction limited imaging in two or three axes rather than just the axial direction. In SIM this fine structure is translated into a lower “beat” frequency, which does pass through the lens.
