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Combined molecular techniques reveal DNA details

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Compiled by BioPhotonics staff

A new sensitive instrument that combines two molecular imaging technologies can provide scientists with detailed insight into dynamic molecular processes.

Two physicists from the University of Illinois have combined their expertise in single-molecule biophysics – fluorescence microscopy and optical traps – to study the binding and unbinding of individual DNA segments to a larger strand. Their findings appeared in Nature Methods on Feb. 20, 2011 (doi: 10.1038/nmeth.1574).


Physicists from the University of Illinois have developed an instrument that uses two molecular imaging techniques to capture individual DNA segments binding and unbinding to a larger strand.


On their own, the two techniques cannot provide the sensitivity needed to image single DNA strands. Although fluorescence microscopy techniques enable researchers to observe proteins as they conform and move, they lack the spatial range to track movement over distances. Optical traps allow scientists to study a protein’s translocation, but not its conformation, making it difficult to know how many proteins or which types are involved.

By combining the two methods, the researchers measured both the protein’s motion – sensitive to translocation as small as one DNA base pair – as well as its conformational changes as it acted. Their findings revealed details about the DNA’s mechanism that was not accessible using the two techniques separately.

The work was supported by the National Science Foundation, National Institutes of Health and the Howard Hughes Medical Institute.
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Published: May 2011
Glossary
fluorescence microscopy
Fluorescence microscopy is a specialized optical imaging technique used in biology, chemistry, and materials science to visualize and study specimens that exhibit fluorescence. Fluorescence is the phenomenon where a substance absorbs light at one wavelength and emits light at a longer wavelength. In fluorescence microscopy, fluorescent dyes or proteins are used to label specific structures or molecules within a sample. The basic principles of fluorescence microscopy involve illuminating the...
nano
An SI prefix meaning one billionth (10-9). Nano can also be used to indicate the study of atoms, molecules and other structures and particles on the nanometer scale. Nano-optics (also referred to as nanophotonics), for example, is the study of how light and light-matter interactions behave on the nanometer scale. See nanophotonics.
AmericasBiophotonicsBioScanDNA bindingDNA unbindingfluorescence microscopyHoward Hughes Medical InstituteIllinoisImagingindividual DNA segmentsMicroscopymolecular imagingmolecular processingnanoNational Institutes of HealthNational Science FoudnationNewsoptical trapsOpticsprotein translocationproteinssingle-molecule biophysicsUniversity of Illinois at Urbana-Champaign

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