Iterative expansion microscopy

作者：Chang, Jae-Byum; Chen, Fei; Yoon, Young-Gyu; Jung, Erica E.; Babcock, Hazen; Kang, Jeong Seuk; Asano, Shoh; Suk, Ho-Jun; Pak, Nikita; Tillberg, Paul W.; Wassie, Asmamaw T.; Cai, Dawen; Boyden, Edward S.^*

来源：Nature Methods, 2017, 14(6): 593-+.

DOI：10.1038/NMETH.4261

• IN-SITU
• FLUORESCENT PROTEINS
• RNA
• LOCALIZATION
• POLYACRYLAMIDE
• FLUOROPHORES
• PERFORMANCE
• ANTIBODIES
• VOLUMES
• TISSUES

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.

• 出版日期2017-6