Towards structural biology with super-resolution microscopy

Nanoscale
Julia MolleDina Grohmann

Abstract

Fluorescence resonance energy transfer (FRET) has been instrumental in determining the structure and dynamics of biomolecules but distances above 8 nanometers are not accessible. However, with the advent and rapid development of super-resolution (SR) microscopy, distances between two fluorescent dyes below 20 nanometers can be resolved, which hitherto has been inaccessible for fluorescence microscopy approaches due to the limited resolving power of an optical imaging system that is determined by the fundamental laws of light diffraction (referred to as the diffraction limit). Therefore, the question arises whether SR microscopy can ultimately close the resolution gap between FRET and the diffraction limit and whether SR microscopy can be employed for the structural interrogation of proteins in the sub-20 nm range? Here, we show that the combination of DNA nanotechnology and single-molecule biochemistry allows the first step towards the investigation of the structural organization of a protein via SR microscopy. Limiting factors and possible future directions for the full implementation of SR microscopy as a structural tool are discussed.

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Citations

Mar 8, 2019·Advanced Healthcare Materials·Divita Mathur, Igor L Medintz

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Methods Mentioned

BETA
optical microscopy
fluorescence resonance
electrophoresis
size exclusion chromatography
gel filtration
atomic
AFM
super-resolution microscopy
fluorescence microscopy
light microscopy

Software Mentioned

ProtParam
GATTAquant
Labview
MINFLUX
DNA
PAINT
JPK Data Processing Software
MATLAB
Gattanalysis

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