title: A microfluidic device for the hydrodynamic immobilisation of living fission yeast cells for super-resolution imaging.
abstract: We describe a microfluidic device designed specifically for the reversible immobilisation ofSchizosaccharomyces pombe(Fission Yeast) cells to facilitate live cell super-resolution microscopy. Photo-Activation Localisation Microscopy (PALM) is used to create detailed super-resolution images within living cells with a modal accuracy of >25 nm in the lateral dimensions. The novel flow design captures and holds cells in a well-defined array with minimal effect on the normal growth kinetics. Cells are held over several hours and can continue to grow and divide within the device during fluorescence imaging.
authors.affiliation: Cambridge Nanoscience Centre, 11 J J Thomson Avenue, Cambridge CB3 0FF, United Kingdom.
authors.forename: Laurence
authors.initials: L
authors.lastname: Bell
authors.affiliation: Cambridge Nanoscience Centre, 11 J J Thomson Avenue, Cambridge CB3 0FF, United Kingdom.
authors.forename: Ashwin
authors.initials: A
authors.lastname: Seshia
authors.affiliation: Department of Biochemistry, 80 Tennis Court Road, Cambridge CB2 1GA, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Lando
authors.affiliation: Department of Biochemistry, 80 Tennis Court Road, Cambridge CB2 1GA, United Kingdom.
authors.forename: Ernest
authors.initials: E
authors.lastname: Laue
authors.affiliation: Department of Chemistry, Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Palayret
authors.affiliation: Department of Chemistry, Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: Steven F
authors.initials: SF
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Lensfield Road, Cambridge CB2 1EW, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Klenerman
keywords: Hydrodynamic trapping; Microfluidics; PALM imaging; Single cell analysis; Single molecule localisation microscopy
references.citation: Lab Chip. 2007 Jun;7(6):740-5
references.pmid: 17538716
references.citation: Lab Chip. 2008 May;8(5):663-71
references.pmid: 18432334
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 27;106(43):18149-54
references.pmid: 19826080
references.citation: Lab Chip. 2004 Apr;4(2):98-103
references.pmid: 15052347
references.citation: Electrophoresis. 2008 Dec;29(24):5055-60
references.pmid: 19130590
references.citation: Biophys J. 2011 Apr 6;100(7):L31-3
references.pmid: 21463569
references.citation: Nat Methods. 2008 Nov;5(11):947-9
references.pmid: 18794860
references.citation: Open Biol. 2012 Jul;2(7):120078
references.pmid: 22870388
references.citation: Langmuir. 2004 Dec 21;20(26):11684-91
references.pmid: 15595798
references.citation: Nat Cell Biol. 2010 Aug;12(8):791-8
references.pmid: 20657594
references.citation: Lab Chip. 2006 Nov;6(11):1445-9
references.pmid: 17066168
references.citation: Proc Natl Acad Sci U S A. 2007 Jan 23;104(4):1146-51
references.pmid: 17227861
references.citation: Methods Mol Biol. 2000;139:209-19
references.pmid: 10840789
references.citation: Annu Rev Phys Chem. 2010;61:345-67
references.pmid: 20055680
references.citation: Nature. 1975 Aug 14;256(5518):547-51
references.pmid: 1165770
references.citation: Curr Protoc Mol Biol. 2010 Oct;Chapter 14:Unit14.20
references.pmid: 20890901
references.citation: Biophys J. 2006 Dec 1;91(11):4258-72
references.pmid: 16980368
references.citation: Exp Cell Res. 1988 Mar;175(1):184-91
references.pmid: 3345800
references.citation: Lab Chip. 2010 Apr 7;10(7):857-63
references.pmid: 20300672
references.citation: J Cell Biol. 2010 Jul 26;190(2):165-75
references.pmid: 20643879
references.citation: Science. 1987 Mar 20;235(4795):1517-20
references.pmid: 3547653