Topographic contrast of ultrathin cryo-sections for correlative super-resolution light and electron microscopy.
Fluorescence microscopy reveals molecular expression at nanometer resolution but lacks ultrastructural context information. This deficit often hinders a clear interpretation of results. Electron microscopy provides this contextual subcellular detail, but protein identification can often be problematic. Correlative light and electron microscopy produces complimentary information that expands our knowledge of protein expression in cells and tissue. Inherent methodological difficulties are however encountered when combining these two very different microscopy technologies. We present a quick, simple and reproducible method for protein localization by conventional and super-resolution light microscopy combined with platinum shadowing and scanning electron microscopy to obtain topographic contrast from the surface of ultrathin cryo-sections. We demonstrate protein distribution at nuclear pores and at mitochondrial and plasma membranes in the extended topographical landscape of tissue.
Center for Microscopy and Image Analysis, University of Zurich, Switzerland.
José María
JM
Mateos
Center for Microscopy and Image Analysis, University of Zurich, Switzerland.
Bruno
B
Guhl
Center for Microscopy and Image Analysis, University of Zurich, Switzerland.
Jana
J
Doehner
Center for Microscopy and Image Analysis, University of Zurich, Switzerland.
Gery
G
Barmettler
Center for Microscopy and Image Analysis, University of Zurich, Switzerland.
Andres
A
Kaech
Center for Microscopy and Image Analysis, University of Zurich, Switzerland.
Urs
U
Ziegler
Methods Mol Med. 2006;121:351-69
16251754
Proc Natl Acad Sci U S A. 2012 Apr 17;109(16):6136-41
22474357
PLoS One. 2013 Oct 25;8(10):e77209
24204771
J Neurosci. 2015 Apr 8;35(14):5792-807
25855189
Proc Natl Acad Sci U S A. 2011 Aug 16;108(33):13546-51
21799113
Microsc Res Tech. 2003 Oct 15;62(3):218-24
14506687
J Cell Biol. 1980 Mar;84(3):560-83
6987244
Nat Methods. 2015 Mar;12(3):215-8, 4 p following 218
25581799
Arch Biochem Biophys. 2015 Sep 1;581:98-110
26072116
Nat Methods. 2014 Mar;11(3):305-8
24464288
J Histochem Cytochem. 2013 Mar;61(3):236-47
23264637
J Struct Biol. 2014 May;186(2):273-82
24704216
Histochem J. 1980 Jul;12(4):381-403
7440248
Nat Methods. 2011 Jan;8(1):80-4
21102453
J Struct Biol. 2015 Jan;189(1):53-61
25448886
Nat Methods. 2015 Jun;12(6):503-13
26020503
J Histochem Cytochem. 2009 Dec;57(12):1103-12
19654103
Methods Cell Biol. 2014;124:363-89
25287850
Elife. 2013 Oct 08;2:e01149
24137544
Nano Lett. 2009 Jun;9(6):2508-10
19459703
Nat Protoc. 2007;2(10):2480-91
17947990
Nat Methods. 2012 Jun 28;9(7):676-82
22743772
J Microsc. 1999 Dec;196(Pt 3):279-87
10594768
PLoS One. 2015 Dec 21;10(12):e0145034
26690057
Kidney Int. 2013 May;83(5):811-24
23447069
Sci Rep. 2015 Mar 31;5:9583
25823571
Microsc Res Tech. 2000 Dec 1;51(5):464-8
11074617
Science. 2015 Aug 28;349(6251):aab3500
26315442
J Ultrastruct Res. 1984 Oct;89(1):65-78
6544882
Microsc Res Tech. 2010 Mar;73(3):215-24
19725102
J Struct Biol. 2011 Jul;175(1):62-72
21473917
Science. 2006 Sep 15;313(5793):1642-5
16902090
PLoS One. 2012;7(6):e38011
22723842
PLoS One. 2012;7(4):e35172
22523574