ABSTRACT

This chapter provides an overview of commonly used super-resolution microscopy techniques, their applications to live-cell imaging, and future directions for this family of techniques. Fluorescence microscopy has been a crucial tool in the advancement of modern cell biology because of its noninvasive nature, compatibility with imaging live samples, and molecule-specific labeling tools. The resolution of light microscopy has a fundamental limit. This is commonly referred to as the diffraction limit, as it arises from the diffraction of light that occurs as it passes through an aperture. Super-resolution microscopy refers to any technique based on optical microscopy that is capable of resolving object separations smaller than allowed by Abbe’s diffraction limit. Structured illumination microscopyis a widefield super-resolution technique that uses the mathematical analysis of a sequence of images capturing a sample fluorescently excited by a patterned illumination to retrieve subdiffraction limit features.