Super-resolution microscopy

Super-resolution microscopy is a family of optical imaging methods that resolves details finer than conventional light microscopy’s diffraction limit by controlling illumination, fluorescence, or image reconstruction.

Stimulated emission depletion microscopy: A fluorescence microscopy method that confines emission to a subdiffraction region using a patterned depletion beam. It narrows the fluorescent spot by suppressing emission around the scan position.
Diffraction limit: A bound on the spatial detail a wave-based imaging system can distinguish because of diffraction. It is the conventional resolution benchmark that super-resolution methods exceed.
Single-molecule tracking: The measurement of individual molecules’ positions over time to infer their movement and interactions. Localization methods can follow molecular motion as well as map fixed structures.
Conventional fluorescence microscopy: Fluorescence imaging that resolves structures within the conventional diffraction-limited range. It provides the baseline resolution and acquisition demands that super-resolution methods surpass.
Localization precision: The uncertainty in estimating a fluorescent emitter’s position from measured photons and background. It sets a central limit on the detail attainable by localization-based imaging.
Single-molecule localization microscopy: A family of methods that reconstructs images from the estimated positions of isolated fluorescent molecules. It groups methods that build super-resolved images through repeated localization.
Point spread function: The image formed by an optical system from an ideal point source. Its width determines how signals from nearby emitters overlap.
Live-cell imaging: Microscopy of living cells over time to observe their structures and processes. Some super-resolution methods reveal cellular dynamics, though photon dose and speed constrain them.
Electron microscopy: Imaging that uses electron beams to resolve structures at much finer scales than visible-light microscopy. It offers higher spatial resolution but differs in contrast, sample preparation, and live imaging.
Phototoxicity: Cellular damage caused by exposure to light during imaging. High illumination and repeated acquisition can compromise live-cell measurements.

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