Expansion microscopy at one nanometer resolution

Hümpfer, Nadja

doi:10.1242/prelights.33191

Cited by 2 publications

(4 citation statements)

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“…It is now a widely used high-density reconstruction algorithm, as highlighted by an important uptake by the scientific community [22][23][24][25] . Since its inception, several adaptations of SRRF have been proposed by the community, such as those based on a combination with other advanced imaging approaches [26][27][28] or on the introduction of additional data preprocessing steps 29 (Supplementary Table 2 provides a summary), highlighting the interest in and potential impact of the method on the imaging community. The positive reception of the original SRRF method can also be attributed to its user-friendly and accessible implementation as a plugin for the Fiji framework 30 .…”

Section: Esrrf Provides High-fidelity Srm Imagesmentioning

confidence: 99%

High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation

Laine,

Heil,

Coelho

et al. 2023

Nat Methods

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Live-cell super-resolution microscopy enables the imaging of biological structure dynamics below the diffraction limit. Here we present enhanced super-resolution radial fluctuations (eSRRF), substantially improving image fidelity and resolution compared to the original SRRF method. eSRRF incorporates automated parameter optimization based on the data itself, giving insight into the trade-off between resolution and fidelity. We demonstrate eSRRF across a range of imaging modalities and biological systems. Notably, we extend eSRRF to three dimensions by combining it with multifocus microscopy. This realizes live-cell volumetric super-resolution imaging with an acquisition speed of ~1 volume per second. eSRRF provides an accessible super-resolution approach, maximizing information extraction across varied experimental conditions while minimizing artifacts. Its optimal parameter prediction strategy is generalizable, moving toward unbiased and optimized analyses in super-resolution microscopy.

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Section: Esrrf Provides High-fidelity Srm Imagesmentioning

confidence: 99%

High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation

Laine,

Heil,

Coelho

et al. 2023

Nat Methods

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“…withstand and perform under the combined sample preparation and imaging protocols. To date, ExM was successfully combined with SIM [34], STED [35], dSTORM [36,37] and fluctuation-based techniques [38,39]. These ExM-SRM protocols rely on extrinsic labelling techniques of the targets using labels with organic dyes, mainly immunofluorescence [34][35][36][37][38][39].…”

Section: Introductionmentioning

confidence: 99%

“…To date, ExM was successfully combined with SIM [34], STED [35], dSTORM [36,37] and fluctuation-based techniques [38,39]. These ExM-SRM protocols rely on extrinsic labelling techniques of the targets using labels with organic dyes, mainly immunofluorescence [34][35][36][37][38][39]. Furthermore, dSTORM uses ionic switching buffers needed for the on-and off-blinking of dyes.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, factors that hinder efficient extrinsic labelling, such as a cell wall or a highly crowded and charged cytosol like in microbial organisms, can complicate labelling [40] and can introduce artefacts [41]. To date, there are only a handful of publications on combined ExM and SRM strategies for microbiology, namely two studies using ExSIM of fungi [16] and bacteria [42], and one has combined ExM with SRRF imaging for viral SARS-Cov-2 particles [38].…”

Section: Introductionmentioning

confidence: 99%

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Combining single-molecule and expansion microscopy in fission yeast to visualize protein structures at the nanostructural level

Vojnovic,

Caspari,

Hoşkan

et al. 2024

Open Biol.

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In this work, we have developed an expansion microscopy (ExM) protocol that combines ExM with photoactivated localization microscopy (ExPALM) for yeast cell imaging, and report a robust protocol for single-molecule and expansion microscopy of fission yeast, abbreviated as SExY. Our optimized SExY protocol retains about 50% of the fluorescent protein signal, doubling the amount obtained compared to the original protein retention ExM (proExM) protocol. It allows for a fivefold, highly isotropic expansion of fission yeast cells, which we carefully controlled while optimizing protein yield. We demonstrate the SExY method on several exemplary molecular targets and explicitly introduce low-abundant protein targets (e.g. nuclear proteins such as cbp1 and mis16, and the centromere-specific histone protein cnp1). The SExY protocol optimizations increasing protein yield could be beneficial for many studies, when targeting low abundance proteins, or for studies that rely on genetic labelling for various reasons (e.g. for proteins that cannot be easily targeted by extrinsic staining or in case artefacts introduced by unspecific staining interfere with data quality).

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Expansion microscopy at one nanometer resolution

Cited by 2 publications

References 0 publications

High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation

High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation

Combining single-molecule and expansion microscopy in fission yeast to visualize protein structures at the nanostructural level

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