Welcome to the website for expansion microscopy protocols.
Here you can find relevant papers, protocols, materials lists, and advice on expansion microscopy (ExM), a new technology for imaging biological specimens with fine detail by physically making them bigger through a chemical process that preserves nanoscale isotropy. It enables super-resolution imaging on a conventional light microscope.
Please send inquiries to
Chang, J.-B., Chen, F., Yoon, Y.-G., Jung, E. E., Babcock H., Kang J.-S., Asano S., Suk H.-J., Pak N., Tillberg P.W., Wassie A., Cai D., Boyden E.S. (2016) Iterative expansion microscopy, Nature Methods
, advance online publication, doi:10.1038/nmeth.4261. [Publisher link] [PDF]
Chen, F.*, Wassie, A.T.*, Cote, A.J., Sinha, A., Alon, S., Asano, S., Daugharthy, E.R., Chang, J.-B., Marblestone, A., Church, G.M., Raj, A., Boyden, E.S. (2016) Nanoscale Imaging of RNA with Expansion Microscopy, Nature Methods
13(8):679-84. (*, co-first authors) [Publisher link] [PDF]
Tillberg, P.W.*, Chen, F.*, Piatkevich, K.D., Zhao, Y., Yu, C.-C., English, B.P., Gao, L., Martorell, A., Suk, H.-J., Yoshida, F., DeGennaro, E.M., Roossien, D.H., Gong, G., Seneviratne, U., Tannenbaum, S.R., Desimone, R., Cai, D., Boyden, E.S. (2016) Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies, Nature Biotechnology
34:987–992. (*, co-first authors) [Publisher link] [PDF]
Chen, F.*, Tillberg, P.W.*, Boyden, E.S. (2015) Expansion Microscopy, Science
347(6221):543-548. (*, equal contribution) [Publisher link] [PDF]
Talk at TED.com, "A new way to study the brain's invisible secrets," filmed June 2016. [Link]
Expansion Technologies, Inc.
is producing kits and providing services for expansion microscopy. Note: if you are using kits from Expansion Technologies, please follow the protocols provided by the company. Financial disclosure: E.S.B. is a co-founder of Expansion Technologies, Inc.; many of the authors of the papers above are co-inventors on one or more patents related to expansion microscopy.
Expansion microscopy (ExM) enables large 3-D volumes to be imaged with nanoscale precision, even on ordinary microscopes. It works by physically magnifying the specimen directly, in contrast to traditional optical magnification methods. The protocols below show you how to label your specimens, anchor key labels or biomolecules to the swellable polymer that you will form evenly throughout specimens, mechanically homogenize samples, and expand the resultant tissue-material composites.
There are currently four versions of ExM -- an RNA-anchoring form of ExM for visualization of RNA with nanoscale precision (ExFISH, Nature Methods 2016), a protein-anchoring form of ExM which enables visualization of genetically encoded fluorophores and antibodies (proExM, Nature Biotechnology 2016), the original DNA oligonucleotide-anchoring form of ExM (ExM 1.0, Science 2015), and a protocol that expands a single sample over and over for added resolution (iExM, Nature Methods 2017) - see the "Explore" page for links to the primary papers.
Detailed protocols are found below. If you are using kits from Expansion Technologies, please follow the protocols provided by the company. Please contact us at email@example.com with questions or suggestions.
ExM 1.0 Materials List, v1.3
This materials list contains an inventory of all the materials needed to carry out the ExM 1.0 protocol (Science
Iterative expansion microscopy, V1.0
A detailed description of how to expand cells or tissues twice, using the iterative expansion (iExM) protocol (Nature Methods
ExFISH Protocol For Cells and Tissues, V1.0
A detailed description of how to expand cells or tissues according to the ExFISH protocol (Nature Methods
proExM Protocol For Tissues, V1.0
A detailed description of how to expand tissues according to the proExM protocol (Nature Biotechnology
proExM Protocol For Cultured Cells, V1.0
A detailed description of how to expand cultured cells according to the proExM protocol (Nature Biotechnology
ExM 1.0 Cultured Cell Protocol, V1.4
A detailed description of how to expand cultured cells according to the ExM 1.0 protocol (Science
ExM 1.0 Brain Slice Protocol, V1.4
A detailed description of how to expand brain slices according to the ExM 1.0 protocol (Science
ExM 1.0 DNA-Antibody Conjugation Protocol, V1.2
How to create ExM 1.0-ready secondary antibodies for the ExM 1.0 protocol (Science
proExM for tissues: gelation demonstration
A videorecording of the gelation and digestion steps relevant to proExM, ExFISH, and other ExM protocols, when applied to tissues.