TY - JOUR
T1 - Magnify is a universal molecular anchoring strategy for expansion microscopy
AU - Klimas, Aleksandra
AU - Gallagher, Brendan R.
AU - Wijesekara, Piyumi
AU - Fekir, Sinda
AU - DiBernardo, Emma F.
AU - Cheng, Zhangyu
AU - Stolz, Donna B.
AU - Cambi, Franca
AU - Watkins, Simon C.
AU - Brody, Steven
AU - Horani, Amjad
AU - Barth, Alison L.
AU - Moore, Christopher I.
AU - Ren, Xi
AU - Zhao, Yongxin
N1 - Funding Information:
We thank A. Gittis and J. Schwenk at Carnegie Mellon University for the donation and preparation of mouse brain tissue. We thank R. E. Campbell at the University of Alberta for his helpful discussion in writing this manuscript. We also thank T. Lee at Carnegie Mellon University for the donation of U2OS cells. This work was supported by Carnegie Mellon University and DSF Charitable Foundation (Y.Z. and X.R.), US Department of Defense DoD VR190139 (Y.Z.), NIH Director’s New Innovator Award DP2 OD025926-01 (Y.Z.), The Kauffman Foundation (Y.Z. and A.L.B.), T32 predoctoral training grant (Biomechanics in Regenerative Medicine, BiRM) from the National Institute of Biomedical Imaging and Bioengineering of NIH (P.W.), NIH RF1 MH114103 (A.L.B.), Air Force Office of Scientific Research AFOSR FA9550-19-1-13022629 (A.L.B.), NeuroNex GR5260228.1001 (C.I.M.), the Training Program for Interactionist Cognitive Neuroscience (iCoN) T32MH115895 (C.I.M) and NIH National Center for Research Resources 1S10RR019003-01 (S.C.W.).
Funding Information:
We thank A. Gittis and J. Schwenk at Carnegie Mellon University for the donation and preparation of mouse brain tissue. We thank R. E. Campbell at the University of Alberta for his helpful discussion in writing this manuscript. We also thank T. Lee at Carnegie Mellon University for the donation of U2OS cells. This work was supported by Carnegie Mellon University and DSF Charitable Foundation (Y.Z. and X.R.), US Department of Defense DoD VR190139 (Y.Z.), NIH Director’s New Innovator Award DP2 OD025926-01 (Y.Z.), The Kauffman Foundation (Y.Z. and A.L.B.), T32 predoctoral training grant (Biomechanics in Regenerative Medicine, BiRM) from the National Institute of Biomedical Imaging and Bioengineering of NIH (P.W.), NIH RF1 MH114103 (A.L.B.), Air Force Office of Scientific Research AFOSR FA9550-19-1-13022629 (A.L.B.), NeuroNex GR5260228.1001 (C.I.M.), the Training Program for Interactionist Cognitive Neuroscience (iCoN) T32MH115895 (C.I.M) and NIH National Center for Research Resources 1S10RR019003-01 (S.C.W.).
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/6
Y1 - 2023/6
N2 - Expansion microscopy enables nanoimaging with conventional microscopes by physically and isotropically magnifying preserved biological specimens embedded in a crosslinked water-swellable hydrogel. Current expansion microscopy protocols require prior treatment with reactive anchoring chemicals to link specific labels and biomolecule classes to the gel. We describe a strategy called Magnify, which uses a mechanically sturdy gel that retains nucleic acids, proteins and lipids without the need for a separate anchoring step. Magnify expands biological specimens up to 11 times and facilitates imaging of cells and tissues with effectively around 25-nm resolution using a diffraction-limited objective lens of about 280 nm on conventional optical microscopes or with around 15 nm effective resolution if combined with super-resolution optical fluctuation imaging. We demonstrate Magnify on a broad range of biological specimens, providing insight into nanoscopic subcellular structures, including synaptic proteins from mouse brain, podocyte foot processes in formalin-fixed paraffin-embedded human kidney and defects in cilia and basal bodies in drug-treated human lung organoids.
AB - Expansion microscopy enables nanoimaging with conventional microscopes by physically and isotropically magnifying preserved biological specimens embedded in a crosslinked water-swellable hydrogel. Current expansion microscopy protocols require prior treatment with reactive anchoring chemicals to link specific labels and biomolecule classes to the gel. We describe a strategy called Magnify, which uses a mechanically sturdy gel that retains nucleic acids, proteins and lipids without the need for a separate anchoring step. Magnify expands biological specimens up to 11 times and facilitates imaging of cells and tissues with effectively around 25-nm resolution using a diffraction-limited objective lens of about 280 nm on conventional optical microscopes or with around 15 nm effective resolution if combined with super-resolution optical fluctuation imaging. We demonstrate Magnify on a broad range of biological specimens, providing insight into nanoscopic subcellular structures, including synaptic proteins from mouse brain, podocyte foot processes in formalin-fixed paraffin-embedded human kidney and defects in cilia and basal bodies in drug-treated human lung organoids.
UR - http://www.scopus.com/inward/record.url?scp=85145322005&partnerID=8YFLogxK
U2 - 10.1038/s41587-022-01546-1
DO - 10.1038/s41587-022-01546-1
M3 - Article
C2 - 36593399
AN - SCOPUS:85145322005
SN - 1087-0156
VL - 41
SP - 858
EP - 869
JO - Nature Biotechnology
JF - Nature Biotechnology
IS - 6
ER -