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2016 ; 3
(6
): 3-6
Nephropedia Template TP
gab.com Text
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English Wikipedia
Enhanced DNA imaging using super-resolution microscopy and simultaneous
single-molecule orientation measurements
#MMPMID27722186
Backer AS
; Lee MY
; Moerner WE
Optica
2016[]; 3
(6
): 3-6
PMID27722186
show ga
Single-molecule orientation measurements provide unparalleled insight into a
multitude of biological and polymeric systems. We report a simple,
high-throughput technique for measuring the azimuthal orientation and rotational
dynamics of single fluorescent molecules, which is compatible with localization
microscopy. Our method involves modulating the polarization of an excitation
laser, and analyzing the corresponding intensities emitted by single dye
molecules and their modulation amplitudes. To demonstrate our approach, we use
intercalating and groove-binding dyes to obtain super-resolved images of
stretched DNA strands through binding-induced turn-on of fluorescence. By
combining our image data with thousands of dye molecule orientation measurements,
we develop a means of probing the structure of individual DNA strands, while also
characterizing dye-DNA interactions. This approach may hold promise as a method
for monitoring DNA conformation changes resulting from DNA-binding proteins.