![]() Recently, however, super-resolution fluorescence imaging techniques have emerged which are able to achieve a resolution of ∼50 nm or better while maintaining the high molecular specificity and live-cell compatibility of conventional fluorescence microscopy. Other biological imaging modalities such as electron microscopy, atomic force microscopy, or near-field techniques that place a detector or imaging probe within approximately one wavelength or less of the emitting dipole, are capable of attaining very high spatial resolution but either have a poor ability to identify the molecules being examined or are primarily limited to the imaging of surface features. This resolution limit is caused by the diffraction of light and is a characteristic of all far-field light microscopies, where far-field here refers to configurations where a detector is placed several wavelengths or further from the emitting dipole. While fluorescence microscopy has revolutionized the visualization of structures and dynamics in biology, it has traditionally faced a resolution limit of about 250 nm laterally and 500 nm axially, such that features closer than these distances are obscured. Its immense popularity arises in large part from the ability to specifically label molecules or structures of interest within a cell and from its ability to study living organisms. Fluorescence microscopy allows researchers to explore the inner workings of fixed or living specimens in order to directly monitor organization and dynamics occurring within a given microenvironment.
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