Expansion Microscopy: Super-Resolution Imaging with Hydrogels

Truckenbrodt, Sven

doi:10.1021/acs.analchem.2c04921

Cited by 18 publications

(15 citation statements)

References 187 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Parallel sample preparaKon should furthermore enable straigh•orward implementaKons of tokPAINT for correlaKve light and electron microscopy [89][90][91][92] . With the rise of expansion microscopy 93 , it would be interesKng to apply tokPAINT to cryosecKons of previously expanded samples, which could have advantages, such as increasing epitope accessibility and reducing linkage errors 94 , but would require prior validaKon of preserving cellular integrity at the nanoscale.…”

Section: Discussionmentioning

confidence: 99%

Cryosectioning-enabled super-resolution microscopy for studying nuclear architecture at the single protein level

Stein,

Ericsson,

Nofal

et al. 2024

Preprint

View full text Add to dashboard Cite

DNA-PAINT combined with total Internal Reflection Fluorescence (TIRF) microscopy enables the highest localization precisions, down to single nanometers in thin biological samples, due to TIRF’s unique method for optical sectioning and attaining high contrast. However, most cellular targets elude the accessible TIRF range close to the cover glass and thus require alternative imaging conditions, affecting resolution and image quality. Here, we address this limitation by applying ultrathin physical cryosectioning in combination with DNA-PAINT. With “tomographic & kinetically-enhanced” DNA-PAINT (tokPAINT), we demonstrate the imaging of nuclear proteins with sub-3 nanometer localization precision, advancing the quantitative study of nuclear organization within fixed cells and mouse tissues at the level of single antibodies. We believe that ultrathin sectioning combined with the versatility and multiplexing capabilities of DNA-PAINT will be a powerful addition to the toolbox of quantitative DNA-based super-resolution microscopy in intracellular structural analyses of proteins, RNA and DNAin situ.

show abstract

Section: Discussionmentioning

confidence: 99%

Cryosectioning-enabled super-resolution microscopy for studying nuclear architecture at the single protein level

Stein,

Ericsson,

Nofal

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…Exciting future possibilities for single AAV particle identification also come from expansion microscopy [ 165 , 166 ]. The principle of expansion microscopy is to expand the sample in an isotropic way typically by a factor 10 up to 100.…”

Section: Discussionmentioning

confidence: 99%

Fluorescence Microscopy in Adeno-Associated Virus Research

Golm,

Hübner,

Müller

2023

Viruses

View full text Add to dashboard Cite

Research on adeno-associated virus (AAV) and its recombinant vectors as well as on fluorescence microscopy imaging is rapidly progressing driven by clinical applications and new technologies, respectively. The topics converge, since high and super-resolution microscopes facilitate the study of spatial and temporal aspects of cellular virus biology. Labeling methods also evolve and diversify. We review these interdisciplinary developments and provide information on the technologies used and the biological knowledge gained. The emphasis lies on the visualization of AAV proteins by chemical fluorophores, protein fusions and antibodies as well as on methods for the detection of adeno-associated viral DNA. We add a short overview of fluorescent microscope techniques and their advantages and challenges in detecting AAV.

show abstract

“…Remarkably versatile, ExM can be directly applied to pre-labelled samples (immuno- or dye-labelling) or preserve the inherent fluorescence of genetically-tagged proteins. In all case, proteins and antibodies are anchored to the gel by an acrylamide-conjugated molecule such as succinimidyl ester of 6-((acryloyl)amino)hexanoic acid (AcX) or methacrylic acid N-hydroxysuccinimidyl ester (MA-NHS) (Truckenbrodt, 2023). The acrylamide gel containing the water-absorbent sodium acrylate is then digested by Proteinase K, which then allows expansion of the gels with water.…”

Section: Introductionmentioning

confidence: 99%

Plant Root Expansion Microscopy (ROOT-ExM): A streamlined super resolution method for plants

Grison,

Maucort,

Dumazel

et al. 2024

Preprint

View full text Add to dashboard Cite

Expansion microscopy (ExM) has revolutionized biological imaging by physically enlarging samples, surpassing the light diffraction limit and enabling nanoscale visualization using standard microscopes. While extensively employed across a wide range of biological samples, its application to plant tissues is sparse. In this work, we present ROOT-ExM, an expansion method suited for stiff and intricate multicellular plant tissues, focusing on the primary root of Arabidopsis thaliana. ROOT-ExM achieves isotropic expansion with a fourfold increase in resolution, enabling super-resolution microscopy comparable to STimulated Emission Depletion (STED) microscopy. Labelling is achieved through immunolocalization, compartment-specific dyes, and native fluorescence preservation, while N-Hydroxysuccinimide (NHS) ester-dye conjugates reveal the ultrastructural context of cells alongside specific labelling. We successfully applied ROOT-ExM to image various cellular structures, including the Golgi apparatus, the endoplasmic reticulum, the cytoskeleton, and wall-embedded structures such as plasmodesmata. When combined with lattice light sheet microscopy (LLSM), ROOT-ExM achieves 3D quantitative analysis of nanoscale cellular process, revealing increased vesicular fusion in close proximity of the cell plate during cell division. Achieving super-resolution fluorescence imaging in plant biology remains a formidable challenge. Our findings underscore that ROOT-ExM provides a remarkable, cost-effective solution to this challenge, paving the way for unprecedented insights into plant cellular subcellular architecture.One sentence summaryROOT-ExM achieves super-resolution expansion microscopy in plants

show abstract

Expansion Microscopy: Super-Resolution Imaging with Hydrogels

Cited by 18 publications

References 187 publications

Cryosectioning-enabled super-resolution microscopy for studying nuclear architecture at the single protein level

Cryosectioning-enabled super-resolution microscopy for studying nuclear architecture at the single protein level

Fluorescence Microscopy in Adeno-Associated Virus Research

Plant Root Expansion Microscopy (ROOT-ExM): A streamlined super resolution method for plants

Contact Info

Product

Resources

About