title: Trapping cells on a stretchable microwell array for single-cell analysis.
abstract: There is a need for a technology that can be incorporated into routine laboratory procedures to obtain a continuous, quantitative, fluorescence-based measurement of the dynamic behaviors of numerous individual living cells in parallel, while allowing other manipulations, such as staining, rinsing, and even retrieval of targeted cells. Here, we report a simple, low-cost microarray platform that can trap cells for dynamic single-cell analysis of mammalian cells. The elasticity of polydimethylsiloxane (PDMS) was utilized to trap tens of thousands of cells on an array. The PDMS microwell array was stretched by a tube through which cells were loaded on the array. Cells were trapped on the array by removal of the tube and relaxation of the PDMS. Once that was accomplished, the cells remained trapped on the array without continuous application of an external force and permitted subsequent manipulations, such as staining, rinsing, imaging, and even isolation of targeted cells. We demonstrate the utility of this platform by multicolor analysis of trapped cells and monitoring in individual cells real-time calcium flux after exposure to the calcium ionophore ionomycin. Additionally, a proof of concept for target cell isolation was demonstrated by using a microneedle to locally deform the PDMS membrane in order to retrieve a particular cell from the array.
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: Yuli
authors.initials: Y
authors.lastname: Wang
authors.forename: Pavak
authors.initials: P
authors.lastname: Shah
authors.forename: Colleen
authors.initials: C
authors.lastname: Phillips
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Sims
authors.forename: Nancy L
authors.initials: NL
authors.lastname: Allbritton
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004867:Equipment Design; D015759:Ionomycin / Q000378:metabolism; D059010:Single-Cell Analysis / Q000191:economics / Q000295:instrumentation*; D046888:Tissue Array Analysis / Q000191:economics / Q000295:instrumentation*
chemical_list: D004129:Dimethylpolysiloxanes; D015759:Ionomycin; C013830:baysilon; D002118:Calcium
references.citation: Lab Chip. 2009 Aug 21;9(16):2381-90
references.pmid: 19636470
references.citation: Lab Chip. 2009 Oct 21;9(20):2965-72
references.pmid: 19789751
references.citation: Anal Chem. 2005 Dec 15;77(24):7976-83
references.pmid: 16351145
references.citation: Lab Chip. 2005 Jan;5(1):49-55
references.pmid: 15616740
references.citation: Anal Chem. 2005 Dec 15;77(24):8050-6
references.pmid: 16351155
references.citation: Proc Natl Acad Sci U S A. 2009 Aug 25;106(34):14195-200
references.pmid: 19617544
references.citation: Curr Protoc Mol Biol. 2010 Oct;Chapter 14:Unit14.20
references.pmid: 20890901
references.citation: Nat Rev Mol Cell Biol. 2003 Jul;4(7):517-29
references.pmid: 12838335
references.citation: Cytometry. 1997 Aug 1;28(4):289-97
references.pmid: 9266748
references.citation: ACS Comb Sci. 2011 Mar 14;13(2):190-5
references.pmid: 21395345
references.citation: Anal Chem. 2005 Sep 1;77(17):5628-34
references.pmid: 16131075
references.citation: Integr Biol (Camb). 2009 Dec;1(11-12):625-34
references.pmid: 20027371
references.citation: Cell. 2010 May 14;141(4):559-63
references.pmid: 20478246
references.citation: Biomaterials. 2006 Jun;27(16):3044-63
references.pmid: 16458351
references.citation: Electrophoresis. 2006 Dec;27(24):5093-100
references.pmid: 17117377
references.citation: Lab Chip. 2008 Jul;8(7):1015-23
references.pmid: 18584072
references.citation: Lab Chip. 2009 Oct 21;9(20):2890-5
references.pmid: 19789740
references.citation: Anal Chim Acta. 2009 Sep 7;649(2):141-57
references.pmid: 19699390
references.citation: Lab Chip. 2010 Dec 21;10(24):3363-72
references.pmid: 20967379
references.citation: Lab Chip. 2011 Jan 7;11(1):79-86
references.pmid: 20957290
references.citation: Biomed Microdevices. 2007 Aug;9(4):573-8
references.pmid: 17484053
references.citation: Lab Chip. 2009 Jan 7;9(1):158-63
references.pmid: 19209349
references.citation: Nature. 2010 Jun 10;465(7299):736-45
references.pmid: 20535203
references.citation: Anal Chem. 2009 Jul 1;81(13):5308-13
references.pmid: 19485404
references.citation: Biosens Bioelectron. 2004 Jul 15;19(12):1765-74
references.pmid: 15198083
references.citation: Science. 2006 Apr 14;312(5771):217-24
references.pmid: 16614209
references.citation: Expert Opin Biol Ther. 2002 Apr;2(4):371-93
references.pmid: 11955276
references.citation: Nat Protoc. 2010 Mar;5(3):491-502
references.pmid: 20203666
references.citation: Lab Chip. 2010 Sep 21;10(18):2411-8
references.pmid: 20664846
references.citation: Lab Chip. 2006 Aug;6(8):995-1000
references.pmid: 16874368
references.citation: Nature. 2003 Aug 14;424(6950):810-6
references.pmid: 12917694
references.citation: Lab Chip. 2007 Apr;7(4):423-40
references.pmid: 17389958
references.citation: Lab Chip. 2006 Jan;6(1):121-6
references.pmid: 16372078
references.citation: Biomaterials. 1999 Dec;20(23-24):2363-76
references.pmid: 10614942
references.citation: Lab Chip. 2010 Nov 7;10(21):2952-8
references.pmid: 20838671
references.citation: Lab Chip. 2006 Nov;6(11):1445-9
references.pmid: 17066168