Super-resolution FRET measurements

Szalai, Alan M.; Zaza, Cecilia; Stefani, Fernando D.

doi:10.1039/d1nr05769c

Cited by 23 publications

(15 citation statements)

References 88 publications

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“…A detailed list of possible FRET applications is beyond the scope of this unit. Extensive reviews provide useful examples of FRET studies (Lerner et al., 2018; Lim, Petersen, Bunz, Simon, & Schindler, 2022; Shrestha et al., 2015; Szabó et al., 2020; Szalai, Zaza, & Stefani, 2021). We call attention to a few novel and interesting applications of new FRET methods such as new modalities of flow cytometry, developments in single molecule FRET (smFRET), FRET combined with high resolution and lifetime imaging microscopy (FLIM), and with molecular dynamics (MD) simulations.…”

Section: Applicationsmentioning

confidence: 99%

Principles of Resonance Energy Transfer

2022

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This unit describes the basic principles of Förster resonance energy transfer (FRET). Beginning with a brief summary of the history of FRET applications, the theory of FRET is introduced in detail using figures to explain all the important parameters of the FRET process. After listing various approaches for measuring FRET efficiency, several pieces of advice are given on choosing the appropriate instrumentation. The unit concludes with a discussion of the limitations of FRET measurements followed by a few examples of the latest FRET applications, including new developments such as spectral flow cytometric FRET, single-molecule FRET, and combinations of FRET with super-resolution or lifetime imaging microscopy and with molecular dynamics simulations.

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Section: Applicationsmentioning

confidence: 99%

Principles of Resonance Energy Transfer

2022

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“…[239,240] Combining FRET with other advanced optical microscopy, including single-molecule super-resolution microscopy, was demonstrated in biophysics community for revealing fine structural details. [72,73] Their extended use in polymers should be feasible. It is also important to recognize an inherent limit of FRET process is that it is only applicable when the donor and acceptor distance is in the range of ≈1-10 nm.…”

Section: Discussionmentioning

confidence: 99%

“…The ability to pair these super-resolution techniques with FRET-based measurement enables more accurate studies of phenomena below the diffraction limit with enhanced understanding at a molecular level. [72,73]…”

Section: Fret Paired With Imagingmentioning

confidence: 99%

Fluorescence Resonance Energy Transfer Measurements in Polymer Science: A Review

Valdez

Robertson

Qiang

2022

Macromol. Rapid Commun.

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Fluorescence resonance energy transfer (FRET) is a non‐invasive characterization method for studying molecular structures and dynamics, providing high spatial resolution at nanometer scale. Over the past decades, FRET‐based measurements are developed and widely implemented in synthetic polymer systems for understanding and detecting a variety of nanoscale phenomena, enabling significant advances in polymer science. In this review, the basic principles of fluorescence and FRET are briefly discussed. Several representative research areas are highlighted, where FRET spectroscopy and imaging can be employed to reveal polymer morphology and kinetics. These examples include understanding polymer micelle formation and stability, detecting guest molecule release from polymer host, characterizing supramolecular assembly, imaging composite interfaces, and determining polymer chain conformations and their diffusion kinetics. Finally, a perspective on the opportunities of FRET‐based measurements is provided for further allowing their greater contributions in this exciting area.

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“…Beam-scanning SML-SSI methods are fully compatible with quantitative fluorescence measurements such as fluorescence lifetime (FLIM), , as demonstrated by p-MINFLUX and RASTMIN . FLIM nanoscopy based on SML-SSI can be potentially applied for super-resolved energy transfer measurements or metal or graphene induced energy transfer methods. − …”

Section: Current and Future Applicationsmentioning

confidence: 99%

Nanometer Resolution Imaging and Tracking of Single Fluorophores by Sequential Structured Illumination

et al. 2022

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Localization of single fluorescent emitters is central for many physicochemical and biophysical measurements at the nanoscale and beyond ensemble averaging. Examples include single-molecule tracking and super-resolution imaging by single-molecule localization microscopy. Among the numerous localization methods available, MINFLUX constituted a breakthrough for achieving a ∼10-fold improvement in resolution over wide-field camera-based approaches, reaching the molecular scale at moderate photon counts. Since then, other related methods have been developed and a common framework has been established for single-molecule localization through sequential structured illumination. Now, fluorescence nanoscopy and tracking of single fluorophores with true nanometric resolution are sitting on solid conceptual and experimental ground. There is an ample scope of questions where they can be applied in the near future to gain new insight into the dynamics and photophysics of natural and synthetic systems at the nanometer scale.

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Super-resolution FRET measurements

Abstract: Recent advances combining FRET and super-resolution techniques are presented. This merger of methods is opening exciting opportunities to study the spatial organization and functional state of (bio)molecules and materials at the nanoscale.

Cited by 23 publications

References 88 publications

Principles of Resonance Energy Transfer

Principles of Resonance Energy Transfer

Fluorescence Resonance Energy Transfer Measurements in Polymer Science: A Review

Nanometer Resolution Imaging and Tracking of Single Fluorophores by Sequential Structured Illumination

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