Single-Molecule High-Resolution Colocalization (SHREC)
Single molecule high resolution colocalization (SHREC) is a technique for fluorescence microscopy that allows precise distances to be measured well below the diffraction limit of light microscopy.
Microscopy is often used to investigate biological systems such as cells and macromolecular complexes, and distances measured between two components are frequently used as a proxy for their functional connection. Over the past decade, the spatial resolution of isolated fluorescent probes has improved immensely (Yildiz et al. 2003; Betzig et al. 2006; Mortensen et al. 2010), requiring that colocalization techniques of two fluorescent probes achieve high resolution.
Historically, Förster resonance energy transfer (FRET) has been used to infer proximity below ∼10 nm in biological samples. At the other end of the spectrum, diffraction-limited light microscopy has allowed distance measurements above ∼250 nm. SHREC measures distances...
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