Subnanometre single-molecule localization, registration and distance measurements

@article{Pertsinidis2010SubnanometreSL,
  title={Subnanometre single-molecule localization, registration and distance measurements},
  author={A. Pertsinidis and Yunxiang Zhang and S. Chu},
  journal={Nature},
  year={2010},
  volume={466},
  pages={647-651}
}
Remarkable progress in optical microscopy has been made in the measurement of nanometre distances. If diffraction blurs the image of a point object into an Airy disk with a root-mean-squared (r.m.s.) size of s =  0.44λ/2NA (∼90 nm for light with a wavelength of λ = 600 nm and an objective lens with a numerical aperture of NA = 1.49), limiting the resolution of the far-field microscope in use to d = 2.4s ≈ 200 nm, additional knowledge about the specimen can be used to great advantage. For… Expand
Localization microscopy: mapping cellular dynamics with single molecules
TLDR
The versatility of the SRFLM technique has great promise for improving the understanding of cell biology at the molecular level as well as investigating questions whose answers lie below the diffraction limit. Expand
Superlocalization spectral imaging microscopy of a multicolor quantum dot complex.
TLDR
A spectral imaging based color nanoscopy which is capable of determining which QD is blinking in the multicolor QD complex through tracking the first-order spectrum, and thus, the distance at tens of nanometers between two QDs is measured. Expand
A microscope calibration protocol for single- molecule microscopy
Single-molecule microscopy allows for the investigation of the dynamics of individual molecules and the visualization of subcellular structures at high spatial resolution. For single-molecule imagingExpand
Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes
TLDR
This work introduces MINFLUX, a concept for localizing photon emitters in space by probing the emitter with a local intensity minimum of excitation light, which minimizes the fluorescence photons needed for high localization precision. Expand
Super-Resolution Imaging Through Stochastic Switching and Localization of Single Molecules: An Overview
The resolution of fluorescence microscopy had traditionally been limited to ~200–300 nm due to the diffraction of light. Recently, this resolution limit has been broken using mainly two classes ofExpand
Three-Dimensional Localization of an Individual Fluorescent Molecule with Angstrom Precision.
TLDR
A cryogenic reflecting microscope with a numerical aperture of 0.99 and an imaging stability of0.05 nm in standard deviation at a temperature of 1.8 K to realize the cryogenic performances is developed by the laboratory. Expand
Ultraprecise single-molecule localization microscopy enables in situ distance measurements in intact cells
TLDR
By overcoming the major bottlenecks in SMLM imaging, it is possible to generate molecular images with nanometer accuracy and conduct distance measurements on the biological relevant length scales. Expand
Optimal 3D single-molecule localization for superresolution microscopy with aberrations and engineered point spread functions
TLDR
An optimal 3D single-molecule localization estimator is presented in a general framework for noisy, aberrated and/or engineered PSF imaging and is shown to be efficient, meaning it reaches the fundamental Cramer–Rao lower bound of x, y, and z localization precision. Expand
Superresolution imaging of biological nanostructures by spectral precision distance microscopy
TLDR
The principles of spectrally assigned localization microscopy (SALM) of biological nanostructures are described, focusing on a special SALM approach, spectral precision distance/position determination microscopy(SPDM) with physically modified fluorochromes (SPDMPhymod), based on high‐precision localization of fluorescent molecules. Expand
Microscope calibration protocol for single-molecule microscopy.
TLDR
A method for calibrating a microscope at the nanometer scale, in the sense of determining optical aberrations as revealed by point source localization errors on the order of nanometers, is proposed. Expand
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 41 REFERENCES
Single-molecule high-resolution imaging with photobleaching.
  • M. Gordon, T. Ha, P. Selvin
  • Chemistry, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 2004
TLDR
This work circumvented the Rayleigh limit and achieved nanometer-scale resolution by measuring the distance between single fluorophores separated by 10-20 nm via attachment to the ends of double-stranded DNA molecules immobilized on a surface. Expand
Ultrahigh-resolution multicolor colocalization of single fluorescent probes.
TLDR
An optical ruler based on ultrahigh-resolution colocalization of single fluorescent probes that relies on the use of two unique families of fluorophores, namely energy-transfer fluorescent beads and semiconductor nanocrystal quantum dots, that can be excited by a single laser wavelength but emit at different wavelengths is described. Expand
Far-field fluorescence microscopy beyond the diffraction limit
A technique has been developed to obtain three-dimensional structural information on a length scale well below the Rayleigh length with conventional far-field optics. By spectrally selecting a singleExpand
High-resolution colocalization of single dye molecules by fluorescence lifetime imaging microscopy.
TLDR
The results presented demonstrate the first high-precision distance measurements between single conventional fluorescent dyes based solely on fluorescence lifetime, and can be used to distinguish, and to measure the distance between, two dye molecules that are less than 30 nm apart. Expand
Nanometer-localized multiple single-molecule fluorescence microscopy.
TLDR
This paper demonstrates nanometer-localized multiple single-molecule (NALMS) fluorescence microscopy by using both centroid localization and photobleaching of the single fluorophores to validate the NALMS microscopy approach. Expand
Ultra-high resolution imaging by fluorescence photoactivation localization microscopy.
TLDR
A new method for fluorescence imaging has been developed that can obtain spatial distributions of large numbers of fluorescent molecules on length scales shorter than the classical diffraction limit, and suggests a means to address a significant number of biological questions that had previously been limited by microscope resolution. Expand
Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission.
The diffraction barrier responsible for a finite focal spot size and limited resolution in far-field fluorescence microscopy has been fundamentally broken. This is accomplished by quenching excitedExpand
Single molecule high-resolution colocalization of Cy3 and Cy5 attached to macromolecules measures intramolecular distances through time.
TLDR
This technique's lower resolution limit lies at the upper resolution limit of single molecule FRET (smFRET) microscopy, and the instrumentation and fluorophores used for SHREC can also be used for smFRET, allowing the two types of measurements to be made interchangeably, covering a wide range of interfluorophore distances. Expand
Combining optical trapping and single-molecule fluorescence spectroscopy: enhanced photobleaching of fluorophores.
TLDR
This work shows that optical trapping with single-molecule fluorescence microscopy is feasible even when the fluorophore is directly attached to the trapped particle, by using optimized optical filters, and indicates that the enhanced photobleaching is caused by the absorption of a visible photon followed by the excited-state absorption of an near-infrared photon. Expand
Interlaced optical force-fluorescence measurements for single molecule biophysics.
TLDR
A novel approach to overcome the dramatic reduction of trap-induced photobleaching effects on the common single molecule fluorescence dye Cy3 by alternately modulating the optical trap and excitation beams to prevent simultaneous exposure of the fluorescent dye. Expand
...
1
2
3
4
5
...