Imaging of unresolved objects, superresolution, and precision of distance measurement with video microscopy.

Publisher Summary This chapter discusses the utility of video microscopy to visualize, resolve, and measure widths and distances to precisions that are conventionally considered to be below the limit of the resolution of the light microscope. In an image-forming system, resolution is generally expressed as a measure of the ability to separate the images of two neighboring object points. In the case of a light microscope, producing diffraction-limited images, the resolution limit is defined as the minimum distance between two self-luminous or incoherently illuminated objects or structures whose diffraction images can visually be distinguished as coming from two points. When the diffraction images of the two points overlap to an extent that they can no longer be distinguished from that of an individual object, the two are said not to be resolved or that the distance is less than the limit of resolution. For microscopy, the contrast boosting ability of video allows the use of the best corrected objective lenses not only for bright field and fluorescence but also for polarized light and differential interference contrast (DIC) microscopy.