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Iterative expansion microscopy
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DOI：10.1038/NMETH.4261.png)
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Abstract
En 中文
We recently developed a method called expansion microscopy, in which preserved biological specimens are physically magnified by embedding them in a densely crosslinked polyelectrolyte gel, anchoring key labels or biomolecules to the gel, mechanically homogenizing the specimen, and then swelling the gel-specimen composite by-4.5x in linear dimension. Here we describe iterative expansion microscopy (iExM), in which a sample is expanded-20x. After preliminary expansion a second swellable polymer mesh is formed in the space newly opened up by the first expansion, and the sample is expanded again. iExM expands biological specimens similar to 4.5 x 4.5, or similar to 20x, and enables similar to 25-nm-resolution imaging of cells and tissues on conventional microscopes. We used iExM to visualize synaptic proteins, as well as the detailed architecture of dendritic spines, in mouse brain circuitry.
Keywords:
IN-SITU
FLUORESCENT PROTEINS
RNA
LOCALIZATION
POLYACRYLAMIDE
FLUOROPHORES
PERFORMANCE
ANTIBODIES
VOLUMES
TISSUES
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