Colocalization of Fluorescent Probes: Accurate and Precise Registration with Nanometer Resolution
نویسندگان
چکیده
منابع مشابه
Colocalization of fluorescent probes: accurate and precise registration with nanometer resolution.
Colocalization of fluorescent probes is commonly used in cell biology to discern the proximity of two proteins in the cell. Considering that the resolution limit of optical microscopy is on the order of 250 nm, there has not been a need for high-resolution colocalization techniques. However, with the advent of higher resolution techniques for cell biology and single-molecule biophysics, colocal...
متن کاملColocalization of Fluorescent Probes
1. Introduction Fluorescence labeling techniques are quite useful to microscopists. Not only can fluorescent probes label sub-cellular structures with a high degree of specificity, they can also do so with a very high degree of sensitivity. This combination of specificity and sensitivity permits observations of cellular events that are not always visible using traditional methodologies, such as...
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Calculation of the centroid of the images of individual fluorescent particles and molecules allows localization and tracking in light microscopes to a precision about an order of magnitude greater than the microscope resolution. The factors that limit the precision of these techniques are examined and a simple equation derived that describes the precision of localization over a wide range of co...
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An optical ruler based on ultrahigh-resolution colocalization of single fluorescent probes is described in this paper. It relies on the use of two unique families of fluorophores, namely energy-transfer fluorescent beads (TransFluoSpheres) and semiconductor nanocrystal quantum dots, that can be excited by a single laser wavelength but emit at different wavelengths. A multicolor sample-scanning ...
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An ultrahigh-resolution colocalization method based on the simultaneous acquisition and analysis of spectrally separated images of the excitation point-spread function of point-like fluorescent probes is reviewed. It is shown that molecular distances can be measured with accuracy better than 10 nm using conventional far-field optics. A detailed account of the methodology, theoretical considerat...
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ژورنال
عنوان ژورنال: Cold Spring Harbor Protocols
سال: 2012
ISSN: 1559-6095
DOI: 10.1101/pdb.top067918