Super-Resolved FRET Imaging by Confocal Fluorescence-Lifetime Single-Molecule Localization Microscopy Article Swipe
Cecilia Zaza
,
Germán Chiarelli
,
Ludovit P. Zweifle
,
Mauricio Pilo‐Pais
,
Evangelos Sisamakis
,
Fabio Barachi
,
Fernando D. Stefani
,
Guillermo P. Acuna
·
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.5281/zenodo.7957337
YOU?
·
· 2023
· Open Access
·
· DOI: https://doi.org/10.5281/zenodo.7957337
FRET-based methods are a special tool for detecting interactions between (bio)molecules and their immediate environment. The spatial distribution of molecular interactions and functional states can be seen using FLIM (Fluorescence Lifetime IMaging) and FRET imaging. The spatial information, accuracy, and dynamic range of the observed signals are, however, constrained by the fact that conventional FLIM and FRET imaging only provides average information over an ensemble of molecules within a diffraction-limited volume. On the other hand, conventional Single Molecule Localization Microscopy (SMLM) relies on highly sensitive multi-pixel detectors (e.g. sCMOS or EM-CCD) whose time resolution is not suitable for fluorescence lifetime measurements. Here, we demonstrate a method for obtaining super-resolved FRET imaging using confocal fluorescence-lifetime single-molecule localization microscopy. The proof of concept was carried out using a DNA origami sample for performing DNA-PAINT measurements in combination with fluorogenic probes for reducing background signal. With this method, We show that FRET events separated by sub-diffraction distances can be distinguished based on lifetime modifications.
Related Topics
Concepts
Förster resonance energy transfer
Confocal
Confocal microscopy
Fluorescence
Fluorescence-lifetime imaging microscopy
Microscopy
Fluorescence microscope
Photoactivated localization microscopy
Biophysics
Chemistry
Materials science
Nuclear magnetic resonance
Optics
Super-resolution microscopy
Physics
Biology
Metadata
- Type
- dataset
- Language
- en
- Landing Page
- https://doi.org/10.5281/zenodo.7957337
- OA Status
- green
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4393966119
All OpenAlex metadata
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- OpenAlex ID
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https://openalex.org/W4393966119Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.5281/zenodo.7957337Digital Object Identifier
- Title
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Super-Resolved FRET Imaging by Confocal Fluorescence-Lifetime Single-Molecule Localization MicroscopyWork title
- Type
-
datasetOpenAlex work type
- Language
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enPrimary language
- Publication year
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2023Year of publication
- Publication date
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2023-05-02Full publication date if available
- Authors
-
Cecilia Zaza, Germán Chiarelli, Ludovit P. Zweifle, Mauricio Pilo‐Pais, Evangelos Sisamakis, Fabio Barachi, Fernando D. Stefani, Guillermo P. AcunaList of authors in order
- Landing page
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https://doi.org/10.5281/zenodo.7957337Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
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greenOpen access status per OpenAlex
- OA URL
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https://doi.org/10.5281/zenodo.7957337Direct OA link when available
- Concepts
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Förster resonance energy transfer, Confocal, Confocal microscopy, Fluorescence, Fluorescence-lifetime imaging microscopy, Microscopy, Fluorescence microscope, Photoactivated localization microscopy, Biophysics, Chemistry, Materials science, Nuclear magnetic resonance, Optics, Super-resolution microscopy, Physics, BiologyTop concepts (fields/topics) attached by OpenAlex
- Cited by
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0Total citation count in OpenAlex
- Related works (count)
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10Other works algorithmically related by OpenAlex
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