Year: 2018
Type: article
Abstract: Expansion microscopy (ExM) is a recently developed technique that enables nanoscale-resolution imaging of preserved cells and tissues on conventional diffraction-limited microscopes via isotropic physical expansion of the specimens before imaging. In ExM, biomolecules and/or fluorescent labels in the specimen are linked to a dense, expandable polymer matrix synthesized evenly throughout the specimen, which undergoes 3-dimensional expansion by ∼4.5 fold linearly when immersed in water. Since our first report, versions of ExM optimized for visualization of proteins, RNA, and other biomolecules have emerged. Here we describe best-practice, step-by-step ExM protocols for performing analysis of proteins (protein retention ExM, or proExM) as well as RNAs (expansion fluorescence in situ hybridization, or ExFISH), using chemicals and hardware found in a typical biology lab. Furthermore, a detailed protocol for handling and mounting expanded samples and for imaging them with confocal and light-sheet microscopes is provided. © 2018 by John Wiley & Sons, Inc.
DOI: 10.1002/cpcb.56
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $5,050
Funders: Howard Hughes Medical Institute, Massachusetts Institute of Technology, Open Philanthropy Project, Cancer Research UK, National Institutes of Health, Intelligence Advanced Research Projects Activity, National Institute of Mental Health, National Human Genome Research Institute, National Institute of Neurological Disorders and Stroke, National Institute of Biomedical Imaging and Bioengineering, Army Research Office, Army Research Laboratory
Awards: G1067232454, Expansion Microscopy, An Accessible Optical Toolbox for Saturated Nanoscale Analysis of Neural Architecture, Millisecond-Timescale Whole-Brain Neural Activity Mapping in Health and Disease, G2645970597, G3030293856, Center for Genomically Engineered Organs, Scalable Cell- and Circuit-Targeted Electrophysiology, G4591667967, Whole Brain Nanoscale Preservation/Imaging, G5197663545, G6960262275, G79353681, G8108977101, G9000073667, G9000125229
OpenAlex ID: W2885194328