title: Cryo-electron tomography: the challenge of doing structural biology in situ.
abstract: Electron microscopy played a key role in establishing cell biology as a discipline, by producing fundamental insights into cellular organization and ultrastructure. Many seminal discoveries were made possible by the development of new sample preparation methods and imaging modalities. Recent technical advances include sample vitrification that faithfully preserves molecular structures, three-dimensional imaging by electron tomography, and improved image-processing methods. These new techniques have enabled the extraction of high fidelity structural information and are beginning to reveal the macromolecular organization of unperturbed cellular environments.
authors.affiliation: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Vladan
authors.initials: V
authors.lastname: Lučič
authors.forename: Alexander
authors.initials: A
authors.lastname: Rigort
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Baumeister
mesh_terms: D020285:Cryoelectron Microscopy; D004247:DNA / Q000737:chemistry*; D021621:Imaging, Three-Dimensional; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D014054:Tomography
chemical_list: D011506:Proteins; D004247:DNA
references.citation: Structure. 2012 Sep 5;20(9):1508-18
references.pmid: 22819217
references.citation: Philos Trans R Soc Lond B Biol Sci. 1971 May 27;261(837):133-8
references.pmid: 4399200
references.citation: Nature. 2007 Oct 4;449(7162):611-5
references.pmid: 17851530
references.citation: Nat Rev Mol Cell Biol. 2006 Mar;7(3):225-30
references.pmid: 16482091
references.citation: EMBO J. 2012 Feb 1;31(3):552-62
references.pmid: 22157822
references.citation: J Ultrastruct Res. 1957 Dec;1(2):109-37
references.pmid: 13492345
references.citation: J Bacteriol. 2009 Aug;191(16):5026-36
references.pmid: 19429612
references.citation: Micron. 2012 Oct;43(10):1010-30
references.pmid: 22658288
references.citation: Mol Microbiol. 2008 Jul;69(1):30-41
references.pmid: 18363791
references.citation: J Struct Biol. 2011 Sep;175(3):372-83
references.pmid: 21616152
references.citation: J Cell Biol. 2009 Aug 10;186(3):437-46
references.pmid: 19667131
references.citation: Trends Parasitol. 2012 Nov;28(11):479-85
references.pmid: 22980758
references.citation: Science. 2012 Aug 3;337(6094):553
references.pmid: 22798403
references.citation: J Struct Biol. 2008 Mar;161(3):459-68
references.pmid: 17923421
references.citation: Nat Methods. 2007 Mar;4(3):215-7
references.pmid: 17277781
references.citation: Exp Cell Res. 1953 Sep;5(1):255-6
references.pmid: 13083636
references.citation: J Struct Biol. 2007 Nov;160(2):146-56
references.pmid: 17905597
references.citation: J Struct Biol. 2010 Apr;170(1):134-45
references.pmid: 20035877
references.citation: J Bacteriol. 2008 Aug;190(16):5672-80
references.pmid: 18567661
references.citation: Structure. 2011 Feb 9;19(2):147-54
references.pmid: 21300284
references.citation: J Struct Biol. 2010 Nov;172(2):180-90
references.pmid: 20638479
references.citation: J Cell Biol. 2009 Dec 14;187(6):921-33
references.pmid: 20008568
references.citation: J Biophys Biochem Cytol. 1961 May;10:1-13
references.pmid: 13772269
references.citation: Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2399-406
references.pmid: 11226251
references.citation: Mol Microbiol. 2009 Mar;71(6):1415-34
references.pmid: 19210619
references.citation: Nat Rev Mol Cell Biol. 2003 Sep;Suppl:SS6-10
references.pmid: 14587520
references.citation: Biochim Biophys Acta. 1978 Sep 29;515(3):303-27
references.pmid: 359049
references.citation: Proc Natl Acad Sci U S A. 2012 Jun 5;109(23):E1481-8
references.pmid: 22556268
references.citation: J Electron Microsc (Tokyo). 2007 Jun;56(3):93-101
references.pmid: 17967812
references.citation: J Cell Biol. 2011 Nov 14;195(4):673-87
references.pmid: 22065640
references.citation: Q Rev Biophys. 1988 May;21(2):129-228
references.pmid: 3043536
references.citation: J Cell Biol. 2010 Aug 23;190(4):613-21
references.pmid: 20733057
references.citation: J Struct Biol. 2003 Oct-Nov;144(1-2):152-61
references.pmid: 14643218
references.citation: J Microsc. 1983 Jul;131(Pt 1):1-9
references.pmid: 6350598
references.citation: J Struct Biol. 2005 Apr;150(1):109-21
references.pmid: 15797735
references.citation: Nat Biotechnol. 2004 Oct;22(10):1317-21
references.pmid: 15470473
references.citation: Proc Soc Exp Biol Med. 1950 Jun;74(2):436-9
references.pmid: 15440846
references.citation: J Struct Biol. 2008 Mar;161(3):276-86
references.pmid: 17720536
references.citation: J Struct Biol. 2011 Dec;176(3):404-8
references.pmid: 21933715
references.citation: Nature. 2003 Jan 23;421(6921):421-2
references.pmid: 12540914
references.citation: Curr Opin Struct Biol. 2013 Apr;23(2):261-7
references.pmid: 23466038
references.citation: Annu Rev Biophys Biomol Struct. 2002;31:303-19
references.pmid: 11988472
references.citation: Trends Cell Biol. 1999 Mar;9(3):92-3
references.pmid: 10201073
references.citation: Ultramicroscopy. 1992 Oct;46(1-4):207-27
references.pmid: 1481272
references.citation: Cell Microbiol. 2010 Mar;12(3):362-71
references.pmid: 19863555
references.citation: J Struct Biol. 1997 Dec;120(3):276-308
references.pmid: 9441933
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 20;109(12):4449-54
references.pmid: 22392984
references.citation: J Cell Biol. 1981 Dec;91(3 Pt 2):IX-XI
references.pmid: 7033234
references.citation: J Biophys Biochem Cytol. 1957 Jan 25;3(1):45-60
references.pmid: 13416310
references.citation: J Struct Biol. 2006 Nov;156(2):334-41
references.pmid: 16857386
references.citation: Science. 1949 Jul 15;110(2846):66-8
references.pmid: 17774956
references.citation: Proc Natl Acad Sci U S A. 2011 Oct 18;108(42):E845-53
references.pmid: 21930914
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3777-81
references.pmid: 17360429
references.citation: Mol Biol Cell. 2012 Jan;23(1):111-20
references.pmid: 22072792
references.citation: J Microsc. 2009 Jul;235(1):1-8
references.pmid: 19566622
references.citation: J Cell Biol. 2006 Sep 11;174(6):759-65
references.pmid: 16954350
references.citation: Structure. 2005 Mar;13(3):423-34
references.pmid: 15766544
references.citation: Science. 2009 Jan 23;323(5913):509-12
references.pmid: 19095899
references.citation: Nature. 2007 Dec 13;450(7172):973-82
references.pmid: 18075576
references.citation: Biol Cell. 2002 Dec;94(9):561-96
references.pmid: 12732437
references.citation: Biol Cell. 2011 Sep;103(9):405-20
references.pmid: 21812762
references.citation: J Struct Biol. 2012 Jan;177(1):135-44
references.pmid: 21907807
references.citation: Mol Cell. 2010 Aug 27;39(4):560-9
references.pmid: 20797628
references.citation: Nature. 2002 Jan 10;415(6868):141-7
references.pmid: 11805826
references.citation: Nature. 2007 Dec 6;450(7171):832-7
references.pmid: 18064004
references.citation: Methods Enzymol. 2010;481:343-69
references.pmid: 20887864
references.citation: Tissue Cell. 2004 Apr;36(2):83-94
references.pmid: 15041410
references.citation: J Struct Biol. 2007 Nov;160(2):135-45
references.pmid: 17884579
references.citation: J Microsc. 2006 Apr;222(Pt 1):42-7
references.pmid: 16734713
references.citation: J Cell Biol. 2010 Aug 23;190(4):491-500
references.pmid: 20733050
references.citation: J Biol Chem. 2011 Aug 19;286(33):29175-29191
references.pmid: 21700706
references.citation: Ultramicroscopy. 2006 May;106(7):587-96
references.pmid: 16616422
references.citation: Nat Cell Biol. 2010 Aug;12(8):739-46
references.pmid: 20639870
references.citation: Annu Rev Biochem. 2005;74:833-65
references.pmid: 15952904
references.citation: J Struct Biol. 2010 Jan;169(1):54-61
references.pmid: 19699799
references.citation: Eur J Cell Biol. 2009 Nov;88(11):669-84
references.pmid: 19726102
references.citation: J Microsc. 2011 Jun;242(3):221-7
references.pmid: 21175615
references.citation: Q Rev Biophys. 2012 Feb;45(1):27-56
references.pmid: 22082691
references.citation: Science. 2004 Feb 6;303(5659):808-13
references.pmid: 14764870
references.citation: Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3963-7
references.pmid: 18316738
references.citation: Nature. 1999 Dec 2;402(6761 Suppl):C47-52
references.pmid: 10591225
references.citation: EMBO J. 2007 Nov 14;26(22):4694-708
references.pmid: 17948052
references.citation: J Cell Biol. 2010 Jan 11;188(1):145-56
references.pmid: 20065095
references.citation: J Struct Biol. 2007 Oct;160(1):70-82
references.pmid: 17692533
references.citation: Histochem Cell Biol. 2008 Nov;130(5):877-89
references.pmid: 18795316
references.citation: Ultramicroscopy. 2009 Aug;109(9):1126-43
references.pmid: 19497671
references.citation: Curr Opin Cell Biol. 2013 Feb;25(1):125-33
references.pmid: 23183140
references.citation: Proc Natl Acad Sci U S A. 2008 Oct 28;105(43):16555-60
references.pmid: 18940922
references.citation: Science. 1974 Dec 13;186(4168):1036-7
references.pmid: 4469695
references.citation: J Struct Biol. 2008 Sep;163(3):271-6
references.pmid: 18588985
references.citation: Structure. 2012 Jun 6;20(6):998-1006
references.pmid: 22632834
references.citation: Science. 2011 Dec 2;334(6060):1283-6
references.pmid: 22075726
references.citation: Science. 2002 Nov 8;298(5596):1209-13
references.pmid: 12424373
references.citation: Methods Cell Biol. 2012;111:259-81
references.pmid: 22857933
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):19732-7
references.pmid: 19064912
references.citation: PLoS Biol. 2004 Nov;2(11):e329
references.pmid: 15514700
references.citation: J Cell Biol. 1979 May;81(2):275-300
references.pmid: 38256
references.citation: Q Rev Biophys. 2004 Feb;37(1):3-13
references.pmid: 17390603
references.citation: Microsc Res Tech. 1998 Jul 1;42(1):43-58
references.pmid: 9712162
references.citation: J Struct Biol. 2013 Mar;181(3):283-90
references.pmid: 23261400
references.citation: J Struct Biol. 2006 Jan;153(1):1-13
references.pmid: 16343943
references.citation: J Struct Biol. 2009 Mar;165(3):157-68
references.pmid: 19114107
references.citation: J Electron Microsc (Tokyo). 2011;60 Suppl 1:S149-59
references.pmid: 21844586
references.citation: Acta Hist Leopoldina. 1979;12:7-136
references.pmid: 11628633
references.citation: Proc Natl Acad Sci U S A. 2012 Mar 6;109(10):3766-71
references.pmid: 22355139
references.citation: PLoS One. 2007 May 09;2(5):e420
references.pmid: 17487274
references.citation: Nat Struct Mol Biol. 2010 Jun;17(6):761-7
references.pmid: 20453857
references.citation: EMBO J. 2011 May 4;30(9):1719-29
references.pmid: 21441899
references.citation: Proc Natl Acad Sci U S A. 2005 Mar 29;102(13):4729-34
references.pmid: 15774580
references.citation: J Struct Biol. 2010 Nov;172(2):169-79
references.pmid: 20178848
references.citation: J Cell Biol. 2013 May 27;201(5):725-40
references.pmid: 23712261
references.citation: Science. 1972 Jul 21;177(4045):268-70
references.pmid: 5041024
references.citation: Microsc Res Tech. 2007 Mar;70(3):230-42
references.pmid: 17279510
references.citation: J Microsc. 2007 Aug;227(Pt 2):98-109
references.pmid: 17845705
references.citation: J Struct Biol. 2011 Feb;173(2):345-9
references.pmid: 20863895
references.citation: Ultramicroscopy. 1984;13(1-2):57-70
references.pmid: 6382732
references.citation: Anat Rec. 1953 Dec;117(4):685-710
references.pmid: 13124776
references.citation: J Cell Comp Physiol. 1953 Aug;42(1):45-70
references.pmid: 13084706
references.citation: Cell. 1998 Feb 6;92(3):291-4
references.pmid: 9476889
references.citation: J Struct Biol. 2001 Sep;135(3):239-50
references.pmid: 11722164
references.citation: Nat Methods. 2009 Nov;6(11):817-23
references.pmid: 19838170
references.citation: J Struct Biol. 2004 Oct;148(1):131-5
references.pmid: 15363793