Expansion microscopy

Chen, Fei; Tillberg, Paul W; Boyden, Edward S.

doi:10.1126/science.1260088

Cited by 1,172 publications

(1,353 citation statements)

References 29 publications

(19 reference statements)

Supporting

Mentioning

1,315

Contrasting

Unclassified

Order By: Relevance

“…The distortions of the sample introduced by the gel during swelling are minimal (Fig 2D) and are virtually identical to those seen in 4× expansion microscopy 7, 8, 12. We would like to note, however, that the extensive digestion required for X10 is incompatible with expansion microscopy protocols that preserve fluorescent proteins 9.…”

Section: Resultssupporting

confidence: 56%

“…This would be ~20–30 nm for normal immunostaining experiments, since these rely on identifying the epitopes via primary antibodies that are later detected through secondary antibodies, each of which is ~10–15 nm in size. Expansion microscopy, a technique introduced by the Boyden laboratory 7, 8, 9, 10, is an important step in this direction. Expansion microscopy entails that the sample of interest is first fixed, permeabilized, and immunostained and is then embedded in polyelectrolyte gels, which expand strongly when dialyzed in water.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

X10 expansion microscopy enables 25‐nm resolution on conventional microscopes

et al. 2018

View full text Add to dashboard Cite

Expansion microscopy is a recently introduced imaging technique that achieves super‐resolution through physically expanding the specimen by ~4×, after embedding into a swellable gel. The resolution attained is, correspondingly, approximately fourfold better than the diffraction limit, or ~70 nm. This is a major improvement over conventional microscopy, but still lags behind modern STED or STORM setups, whose resolution can reach 20–30 nm. We addressed this issue here by introducing an improved gel recipe that enables an expansion factor of ~10× in each dimension, which corresponds to an expansion of the sample volume by more than 1,000‐fold. Our protocol, which we termed X10 microscopy, achieves a resolution of 25–30 nm on conventional epifluorescence microscopes. X10 provides multi‐color images similar or even superior to those produced with more challenging methods, such as STED, STORM, and iterative expansion microscopy (iExM). X10 is therefore the cheapest and easiest option for high‐quality super‐resolution imaging currently available. X10 should be usable in any laboratory, irrespective of the machinery owned or of the technical knowledge.

show abstract

Section: Resultssupporting

confidence: 56%

Section: Introductionmentioning

confidence: 99%

X10 expansion microscopy enables 25‐nm resolution on conventional microscopes

et al. 2018

View full text Add to dashboard Cite

show abstract

“…ability to systematically map the static distribution of RNAs in situ becomes available (34,35), the dynamic mapping and control of RNAs to assess their causal role in cellular processes such as those explored here. Pumby was able to support specific binding, with sequences differing by as few as two or three bases resulting in less, or even functionally zero, binding.…”

Section: Discussionmentioning

confidence: 99%

Programmable RNA-binding protein composed of repeats of a single modular unit

Adamala

Martin-Alarcon

Boyden

2016

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

The ability to monitor and perturb RNAs in living cells would benefit greatly from a modular protein architecture that targets unmodified RNA sequences in a programmable way. We report that the RNA-binding protein PumHD (Pumilio homology domain), which has been widely used in native and modified form for targeting RNA, can be engineered to yield a set of four canonical protein modules, each of which targets one RNA base. These modules (which we call Pumby, for Pumilio-based assembly) can be concatenated in chains of varying composition and length, to bind desired target RNAs. The specificity of such Pumby-RNA interactions was high, with undetectable binding of a Pumby chain to RNA sequences that bear three or more mismatches from the target sequence. We validate that the Pumby architecture can perform RNA-directed protein assembly and enhancement of translation of RNAs. We further demonstrate a new use of such RNA-binding proteins, measurement of RNA translation in living cells. Pumby may prove useful for many applications in the measurement, manipulation, and biotechnological utilization of unmodified RNAs in intact cells and systems.

show abstract

“…Governments in many countries have prioritized developing a better understanding of the brain, most famously in the United States and Europe by funding the US Brain Initiative (Insel et al 2013) and the EU Human Brain Project (Markram 2012). Simultaneously, new methods such as CLARITY (Chung and Deisseroth 2013) and Expansion Microscopy (Chen et al 2015) allow imaging the brain in novel ways. Automated large-scale morphology reconstruction (Kasthuri et al 2015) will extract neuron morphologies from the images and in addition will provide insight into cell types and possible connectivity within the local microcircuit.…”

Section: Modeldb and The Future Of Neurosciencementioning

confidence: 99%

Twenty years of ModelDB and beyond: building essential modeling tools for the future of neuroscience

et al. 2016

View full text Add to dashboard Cite

Neuron modeling may be said to have originated with the Hodgkin and Huxley action potential model in 1952 and Rall's models of integrative activity of dendrites in 1964. Over the ensuing decades, these approaches have led to a massive development of increasingly accurate and complex data-based models of neurons and neuronal circuits. ModelDB was founded in 1996 to support this new field and enhance the scientific credibility and utility of computational neuroscience models by providing a convenient venue for sharing them. It has grown to include over 1100 published models covering more than 130 research topics. It is actively curated and developed to help researchers discover and understand models of interest. ModelDB also provides mechanisms to assist running models both locally and remotely, and has a graphical tool that enables users to explore the anatomical and biophysical properties that are represented in a model. Each of its capabilities is undergoing continued refinement and improvement in response to user experience. Large research groups (Allen Brain Institute, EU Human Brain Project, etc.) are emerging that collect data across multiple scales and integrate that data into many complex models, presenting new challenges of scale. We end by predicting a future for neuroscience increasingly fueled by new technology and high performance computation, and increasingly in need of comprehensive user-friendly databases such as ModelDB to provide the means to integrate the data for deeper insights into brain function in health and disease.

show abstract

Expansion microscopy

Cited by 1,172 publications

References 29 publications

X10 expansion microscopy enables 25‐nm resolution on conventional microscopes

X10 expansion microscopy enables 25‐nm resolution on conventional microscopes

Programmable RNA-binding protein composed of repeats of a single modular unit

Twenty years of ModelDB and beyond: building essential modeling tools for the future of neuroscience

Contact Info

Product

Resources

About