Abstract
The microscope allows scientists to peer into small objects. With new organic dyes and fluorescent proteins, the microscope has improved our ability to extract biological information from selectively labeled proteins, DNA, and organelles. However, a physical property of light, called diffraction, has made it nearly impossible to see objects smaller than the diffraction limit, which is about 250 nm. However, much information smaller than this distance can be insightful. This can be accomplished without violating the diffraction limit using a technique called FIONA (Fluorescence Imaging with One Nanometer Accuracy). With FIONA, the position of a diffraction-limited spot can be located to within a few nanometers, on a time-scale longer than a millisecond. In this chapter, we discuss some background principles underlying FIONA and its applications.
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