title: Correlation of dual colour single particle trajectories for improved detection and analysis of interactions in living cells.
abstract: Interactions between objects inside living cells are often investigated by looking for colocalization between fluorescence microscopy images that are recorded in separate colours corresponding to the fluorescent label of each object. The fundamental limitation of this approach in the case of dynamic objects is that coincidental colocalization cannot be distinguished from true interaction. Instead, correlation between motion trajectories obtained by dual colour single particle tracking provides a much stronger indication of interaction. However, frequently occurring phenomena in living cells, such as immobile phases or transient interactions, can limit the correlation to small parts of the trajectories. The method presented here, developed for the detection of interaction, is based on the correlation inside a window that is scanned along the trajectories, covering different subsets of the positions. This scanning window method was validated by simulations and, as an experimental proof of concept, it was applied to the investigation of the intracellular trafficking of polymeric gene complexes by endosomes in living retinal pigment epithelium cells, which is of interest to ocular gene therapy.
authors: Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Harelbekestraat 72, B-9000 Gent, Belgium. hendrik.deschout@ugent.be
authors: Hendrik
authors: H
authors: Deschout
authors: Thomas
authors: T
authors: Martens
authors: Dries
authors: D
authors: Vercauteren
authors: Katrien
authors: K
authors: Remaut
authors: Jo
authors: J
authors: Demeester
authors: Stefaan C
authors: SC
authors: De Smedt
authors: Kristiaan
authors: K
authors: Neyts
authors: Kevin
authors: K
authors: Braeckmans
mesh_terms: D000465:Algorithms; D002460:Cell Line; D003198:Computer Simulation; D011992:Endosomes / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D008954:Models, Biological; D050997:Nanomedicine; D053758:Nanoparticles / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes
references: Nat Biotechnol. 2003 Nov;21(11):1387-95
references: 14595367
references: Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9461-6
references: 10931959
references: Annu Rev Biophys Biomol Struct. 1997;26:373-99
references: 9241424
references: Biophys J. 2005 Jan;88(1):623-38
references: 15533928
references: Biophys J. 2002 May;82(5):2775-83
references: 11964263
references: Histochem Cell Biol. 2012 Jan;137(1):1-10
references: 22086768
references: Anal Chem. 2001 Mar 15;73(6):1100-5
references: 11305637
references: Biophys J. 2004 Feb;86(2):1185-200
references: 14747353
references: J Neurosci. 2004 Apr 21;24(16):4070-81
references: 15102922
references: J Struct Biol. 2007 Sep;159(3):347-58
references: 17532228
references: Biophys J. 2010 Sep 22;99(6):1996-2005
references: 20858446
references: Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1419-23
references: 15668396
references: J Microsc. 2003 Nov;212(Pt 2):122-31
references: 14629561
references: J Cell Sci. 1998 Jan;111 ( Pt 2):271-81
references: 9405317
references: Mol Ther. 2007 Jul;15(7):1297-305
references: 17457321
references: Biophys J. 2013 Jun 4;104(11):2373-82
references: 23746509
references: J Control Release. 2007 Aug 28;121(3):208-17
references: 17630013
references: Biophys J. 2005 Mar;88(3):2126-36
references: 15596511
references: Angew Chem Int Ed Engl. 2006 Feb 27;45(10):1568-72
references: 16470762
references: Biophys J. 2003 Dec;85(6):4110-21
references: 14645098
references: Biophys J. 2006 Dec 15;91(12):4611-22
references: 17012312
references: Biophys J. 2004 Jun;86(6):3993-4003
references: 15189895
references: Am J Physiol Cell Physiol. 2011 Apr;300(4):C723-42
references: 21209361
references: J Microsc. 2000 Oct;200(Pt 1):14-25
references: 11012824
references: Biophys J. 2009 Aug 19;97(4):L5-7
references: 19686638
references: J Microsc. 2006 Dec;224(Pt 3):213-32
references: 17210054
references: Acta Histochem Cytochem. 2007 Aug 30;40(4):101-11
references: 17898874
references: Proc Natl Acad Sci U S A. 2008 Mar 4;105(9):3298-303
references: 18305159
references: J Biophotonics. 2012 Jan;5(1):97-109
references: 22083848
references: J Microsc. 2008 Apr;230(Pt 1):121-33
references: 18387047
references: Nature. 2010 Jul 29;466(7306):647-51
references: 20613725
references: ACS Nano. 2011 Oct 25;5(10):7874-84
references: 21923168
references: J Microsc. 1993 Mar;169(3):375-382
references: 33930978