title: Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging.
abstract: Fluorescence microscopy is used extensively in cell-biological and biomedical research, but it is often plagued by three major problems with the presently available fluorescent probes: photobleaching, blinking, and large size. We have addressed these problems, with special attention to single-molecule imaging, by developing biocompatible, red-emitting silicon nanocrystals (SiNCs) with a 4.1-nm hydrodynamic diameter. Methods for producing SiNCs by simple chemical etching, for hydrophilically coating them, and for conjugating them to biomolecules precisely at a 1:1 ratio have been developed. Single SiNCs neither blinked nor photobleached during a 300-min overall period observed at video rate. Single receptor molecules in the plasma membrane of living cells (using transferrin receptor) were imaged for ≥10 times longer than with other probes, making it possible for the first time to observe the internalization process of receptor molecules at the single-molecule level. Spatial variations of molecular diffusivity in the scale of 1-2 µm, i.e., a higher level of domain mosaicism in the plasma membrane, were revealed.
authors.affiliation: Institute for Integrated Cell-Material Sciences, 2 Institute for Frontier Medical Sciences, and 3 Department of Materials Science and Engineering, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: Hirohito
authors.initials: H
authors.lastname: Nishimura
authors.forename: Ken
authors.initials: K
authors.lastname: Ritchie
authors.forename: Rinshi S
authors.initials: RS
authors.lastname: Kasai
authors.forename: Miki
authors.initials: M
authors.lastname: Goto
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Morone
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sugimura
authors.forename: Koichiro
authors.initials: K
authors.lastname: Tanaka
authors.forename: Ichiro
authors.initials: I
authors.lastname: Sase
authors.forename: Akihiko
authors.initials: A
authors.lastname: Yoshimura
authors.forename: Yoshitaro
authors.initials: Y
authors.lastname: Nakano
authors.forename: Takahiro K
authors.initials: TK
authors.lastname: Fujiwara
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kusumi
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry*; D002462:Cell Membrane / Q000378:metabolism; D005456:Fluorescent Dyes; D006801:Humans; D008422:Materials Testing; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D057054:Molecular Imaging; D053758:Nanoparticles / Q000737:chemistry*; D036103:Nanotechnology; D038761:Photobleaching; D011990:Receptors, Transferrin / Q000032:analysis* / Q000378:metabolism; D012825:Silicon / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D005456:Fluorescent Dyes; D011990:Receptors, Transferrin; D012825:Silicon
references.citation: Nature. 1997 Jul 24;388(6640):355-8
references.pmid: 9237752
references.citation: Biophys Chem. 2007 Apr;127(1-2):51-63
references.pmid: 17234323
references.citation: Bioconjug Chem. 2011 Jun 15;22(6):1081-8
references.pmid: 21473652
references.citation: Annu Rev Biophys. 2008;37:265-88
references.pmid: 18573082
references.citation: Biophys J. 2008 Jul;95(1):435-50
references.pmid: 18339737
references.citation: J Photochem Photobiol B. 1988 Dec;2(4):523-31
references.pmid: 3150004
references.citation: Biophys J. 2008 Mar 1;94(5):1826-35
references.pmid: 17921203
references.citation: Acc Chem Res. 2010 Feb 16;43(2):190-200
references.pmid: 19827808
references.citation: Biophys J. 1993 Nov;65(5):2021-40
references.pmid: 8298032
references.citation: ACS Nano. 2012 Dec 21;6(12):11080-7
references.pmid: 23181687
references.citation: Annu Rev Biophys Biomol Struct. 2005;34:351-78
references.pmid: 15869394
references.citation: Nature. 2009 Jun 4;459(7247):686-9
references.pmid: 19430463
references.citation: J Cell Biol. 2011 Feb 7;192(3):463-80
references.pmid: 21300851
references.citation: Nat Methods. 2011 Nov 13;9(1):68-71
references.pmid: 22081126
references.citation: Angew Chem Int Ed Engl. 2008;47(29):5465-9
references.pmid: 18601270
references.citation: Protein Expr Purif. 2005 May;41(1):121-7
references.pmid: 15802229
references.citation: Nat Mater. 2008 Aug;7(8):659-64
references.pmid: 18568030
references.citation: Nat Methods. 2008 May;5(5):397-9
references.pmid: 18425138
references.citation: Annu Rev Phys Chem. 2012;63:595-617
references.pmid: 22404588
references.citation: Bioconjug Chem. 2003 Jan-Feb;14(1):195-204
references.pmid: 12526709
references.citation: Biochem Biophys Res Commun. 2001 Jan 12;280(1):312-8
references.pmid: 11162516
references.citation: Biophys J. 1995 Sep;69(3):989-93
references.pmid: 8519998
references.citation: Nat Methods. 2012 Apr 27;9(5):428-9
references.pmid: 22543373
references.citation: Biophys J. 2007 May 15;92(10):3706-18
references.pmid: 17293395
references.citation: Science. 2008 Apr 11;320(5873):201-5
references.pmid: 18403705
references.citation: J Cell Biol. 2002 Jun 10;157(6):1071-81
references.pmid: 12058021
references.citation: Nature. 2010 Apr 1;464(7289):783-7
references.pmid: 20208517
references.citation: Comb Chem High Throughput Screen. 2007 Jul;10(6):473-9
references.pmid: 17896943
references.citation: Bioconjug Chem. 2004 Mar-Apr;15(2):409-12
references.pmid: 15025539
references.citation: ACS Nano. 2008 May;2(5):873-8
references.pmid: 19206483
references.citation: Nat Nanotechnol. 2010 Dec;5(12):878-84
references.pmid: 21113157
references.citation: Langmuir. 2004 May 25;20(11):4720-7
references.pmid: 15969188
references.citation: Biophys J. 2005 Mar;88(3):2126-36
references.pmid: 15596511
references.citation: Science. 2002 May 3;296(5569):913-6
references.pmid: 11988576
references.citation: Biophys J. 2001 Jun;80(6):2667-77
references.pmid: 11371443
references.citation: Science. 2012 Jun 29;336(6089):1676-81
references.pmid: 22745423
references.citation: Annu Rev Cell Dev Biol. 2010;26:285-314
references.pmid: 20929313
references.citation: Biochem Biophys Res Commun. 2006 Dec 8;351(1):7-13
references.pmid: 17055452
references.citation: J Am Chem Soc. 2010 Apr 7;132(13):4556-7
references.pmid: 20235547
references.citation: ACS Nano. 2011 Jan 25;5(1):413-23
references.pmid: 21138323
references.citation: J Biol Chem. 2000 Dec 8;275(49):38135-8
references.pmid: 11027676
references.citation: Sci STKE. 2006 Mar 21;2006(327):pe13
references.pmid: 16552090
references.citation: Proc Natl Acad Sci U S A. 2005 Oct 4;102(40):14284-9
references.pmid: 16169907
references.citation: Arch Biochem Biophys. 1967 Mar;119(1):41-9
references.pmid: 6052434
references.citation: Biochem J. 2011 Feb 1;433(3):411-22
references.pmid: 21235524
references.citation: Bioconjug Chem. 1993 Mar-Apr;4(2):105-11
references.pmid: 7873641
references.citation: Phys Rev Lett. 2005 Mar 4;94(8):087405
references.pmid: 15783932
references.citation: Biophys J. 2007 Apr 15;92(8):2926-43
references.pmid: 17259276
references.citation: Bioconjug Chem. 2008 Mar;19(3):680-5
references.pmid: 18290602
references.citation: Neuron. 2007 Mar 1;53(5):719-34
references.pmid: 17329211
references.citation: Angew Chem Int Ed Engl. 2005 Jul 18;44(29):4550-4
references.pmid: 15973756
references.citation: Proc Natl Acad Sci U S A. 2004 May 11;101(19):7317-22
references.pmid: 15123831
references.citation: J Biomed Mater Res. 1971 Sep;5(5):459-88
references.pmid: 5120386
references.citation: Traffic. 2007 Jun;8(6):702-17
references.pmid: 17461795
references.citation: J Cell Biol. 1990 Feb;110(2):283-94
references.pmid: 2298808
references.citation: J Am Chem Soc. 2008 Apr 16;130(15):5026-7
references.pmid: 18355011