Cryogenic localization of single molecules with angstrom precision

Weisenburger, S.; Jing, Bo; Renn, Alois; Sandoghdar, Vahid

doi:10.1117/12.2025373

Cited by 31 publications

(33 citation statements)

References 36 publications

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“…[6,7] Weisenburger et al compared the performance of dyes with an emission peak of 552-576 nm at liquid helium temperatures (4.4 K), finding an increase of one to two orders of magnitude in the number of detected photons. However, another way is to increase the number of detected photons N. Thus, by cooling a sample and thereby increasing the photostability of the fluorescent dye molecules can have a tremendous impact on resolution in SMLM.…”

Section: Introductionmentioning

confidence: 99%

Photon Yield Enhancement of Red Fluorophores at Cryogenic Temperatures

Hulleman

Gregor

et al. 2018

ChemPhysChem

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Single Molecule Localization Microscopy has become one of the most successful and widely applied methods of Super-resolution Fluorescence Microscopy. Its achievable resolution strongly depends on the number of detectable photons from a single molecule until photobleaching. By cooling a sample from room temperature down to liquid nitrogen temperatures, the photostability of dyes can be enhanced by more than 100 fold, which results in an improvement in localization precision greater than 10 times. Here, we investigate a variety of fluorescent dyes in the red spectral region, and we find an average photon yield between 3.5 ⋅ 10 to 11 ⋅ 10 photons before bleaching at liquid nitrogen temperatures, corresponding to a theoretical localization precision around 0.1 nm.

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Section: Introductionmentioning

confidence: 99%

Photon Yield Enhancement of Red Fluorophores at Cryogenic Temperatures

Hulleman

Gregor

et al. 2018

ChemPhysChem

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“…To improve on this limitation, several efforts have optimized the choices of fluorophores and the buffer conditions [13,135], engineered the dye molecule itself [14], or carefully controlled its environment [136]. The best localization precision for single molecules that has been reported is just under 0.3 nanometers [127].…”

Section: Single Molecule Localizationmentioning

confidence: 99%

Light microscopy: an ongoing contemporary revolution

Weisenburger

Sandoghdar

2015

Contemporary Physics

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Optical microscopy is one of the oldest scientific instruments that is still used in forefront research. Ernst Abbe's nineteenth century formulation of the resolution limit in microscopy let generations of scientists believe that optical studies of individual molecules and resolving sub-wavelength structures were not feasible. The Nobel Prize in 2014 for superresolution fluorescence microscopy marks a clear recognition that the old beliefs have to be revisited. In this article, we present a critical overview of various recent developments in optical microscopy. In addition to the popular super-resolution fluorescence methods, we discuss the prospects of various other techniques and imaging contrasts and consider some of the fundamental and practical challenges that lie ahead.

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“…Thus, by cooling a sample and thereby increasing the photostability of the fluorescent dye molecules can have a tremendous impact on resolution in SMLM. Cooling reduces all photochemical reaction rates and thus inhibits bleaching, thereby enabling significantly more photons to be detected from a molecule, resulting in a localization precision in theÅngström range [6,7]. Weisenburger et al compared the performance of dyes with an emission peak of 552-576 nm at liquid helium temperatures (4.4 K), finding an increase of one to two orders of magnitude in the number of emitted photons [6].…”

Section: Introductionmentioning

confidence: 99%

“…Cooling reduces all photochemical reaction rates and thus inhibits bleaching, thereby enabling significantly more photons to be detected from a molecule, resulting in a localization precision in theÅngström range [6,7]. Weisenburger et al compared the performance of dyes with an emission peak of 552-576 nm at liquid helium temperatures (4.4 K), finding an increase of one to two orders of magnitude in the number of emitted photons [6]. Li et al built a cryo-fluorescence microscope with samples contained in a liquid nitrogen cryostat and analyzed the performance of ATTO 647N, finding an increase of more than two orders of magnitude in photon yield [7].…”

Section: Introductionmentioning

confidence: 99%

Photon yield enhancement of red fluorophores at cryogenic temperatures

Hulleman

Gregor

et al. 2018

Preprint

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Single Molecule Localization Microscopy has become one of the most successful and widely applied methods of Super‐resolution Fluorescence Microscopy. Its achievable resolution strongly depends on the number of detectable photons from a single molecule until photobleaching. By cooling a sample from room temperature down to liquid nitrogen temperatures, the photostability of dyes can be enhanced by more than 100 fold, which results in an improvement in localization precision greater than 10 times. Here, we investigate a variety of fluorescent dyes in the red spectral region, and we find an average photon yield between 3.5 ⋅ 106 to 11 ⋅ 106 photons before bleaching at liquid nitrogen temperatures, corresponding to a theoretical localization precision around 0.1 nm.

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Cryogenic localization of single molecules with angstrom precision

Cited by 31 publications

References 36 publications

Photon Yield Enhancement of Red Fluorophores at Cryogenic Temperatures

Photon Yield Enhancement of Red Fluorophores at Cryogenic Temperatures

Light microscopy: an ongoing contemporary revolution

Photon yield enhancement of red fluorophores at cryogenic temperatures

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