title: Convection-Enhanced Biopatterning with Recirculation of Hydrodynamically Confined Nanoliter Volumes of Reagents.
abstract: We present a new methodology for efficient and high-quality patterning of biological reagents for surface-based biological assays. The method relies on hydrodynamically confined nanoliter volumes of reagents to interact with the substrate at the micrometer-length scale. We study the interplay between diffusion, advection, and surface chemistry and present the design of a noncontact scanning microfluidic device to efficiently present reagents on surfaces. By leveraging convective flows, recirculation, and mixing of a processing liquid, this device overcomes limitations of existing biopatterning approaches, such as passive diffusion of analytes, uncontrolled wetting, and drying artifacts. We demonstrate the deposition of analytes, showing a 2- to 5-fold increase in deposition rate together with a 10-fold reduction in analyte consumption while ensuring less than 6% variation in pattern homogeneity on a standard biological substrate. In addition, we demonstrate the recirculation of a processing liquid using a microfluidic probe (MFP) in the context of a surface assay for (i) probing 12 independent areas with a single microliter of processing liquid and (ii) processing a 2 mm(2) surface to create 170 antibody spots of 50 × 100 μm(2) area using 1.6 μL of liquid. We observe high pattern quality, conservative usage of reagents, micrometer precision of localization and convection-enhanced fast deposition. Such a device and method may facilitate quantitative biological assays and spur the development of the next generation of protein microarrays.
authors.affiliation: IBM Research-Zurich , Säumerstrasse 4, 8803 Rüschlikon, Switzerland.
authors.forename: Julien
authors.initials: J
authors.lastname: Autebert
authors.affiliation: IBM Research-Zurich , Säumerstrasse 4, 8803 Rüschlikon, Switzerland.
authors.forename: Julien F
authors.initials: JF
authors.lastname: Cors
authors.affiliation: IBM Research-Zurich , Säumerstrasse 4, 8803 Rüschlikon, Switzerland.
authors.forename: David P
authors.initials: DP
authors.lastname: Taylor
authors.affiliation: IBM Research-Zurich , Säumerstrasse 4, 8803 Rüschlikon, Switzerland.
authors.forename: Govind V
authors.initials: GV
authors.lastname: Kaigala
mesh_terms: D057446:Hydrodynamics; D007202:Indicators and Reagents / Q000737:chemistry*; D044085:Microfluidics / Q000295:instrumentation; D036103:Nanotechnology
chemical_list: D007202:Indicators and Reagents
references.citation: Proc Natl Acad Sci U S A. 2008 Nov 4;105(44):16843-8
references.pmid: 18974218
references.citation: Chem Commun (Camb). 2009 Jan 21;(3):334-6
references.pmid: 19209320
references.citation: Rev Sci Instrum. 2014 Mar;85(3):034301
references.pmid: 24689601
references.citation: Expert Rev Proteomics. 2011 Feb;8(1):61-79
references.pmid: 21329428
references.citation: Science. 1997 May 2;276(5313):779-81
references.pmid: 9115199
references.citation: Nat Mater. 2007 Oct;6(10):782-9
references.pmid: 17676047
references.citation: Chem Biol. 2001 Feb;8(2):105-15
references.pmid: 11251285
references.citation: Science. 2000 Sep 8;289(5485):1760-3
references.pmid: 10976071
references.citation: Biosensors (Basel). 2014 Apr 24;4(2):111-36
references.pmid: 25587413
references.citation: Langmuir. 2014 Apr 1;30(12):3640-5
references.pmid: 24625080
references.citation: J Am Chem Soc. 2002 Jul 31;124(30):8810-1
references.pmid: 12137530
references.citation: Langmuir. 2011 May 3;27(9):5686-93
references.pmid: 21476506
references.citation: Nat Mater. 2005 Aug;4(8):622-8
references.pmid: 16041377
references.citation: Lab Chip. 2003 Feb;3(1):46-50
references.pmid: 15100805
references.citation: Analyst. 2014 Mar 21;139(6):1303-26
references.pmid: 24479125
references.citation: J Phys Chem B. 2006 Apr 13;110(14):7090-4
references.pmid: 16599468
references.citation: Anal Chem. 2008 Mar 1;80(5):1763-9
references.pmid: 18237157
references.citation: Immunology. 1989 Mar;66(3):321-7
references.pmid: 2649437
references.citation: Science. 1989 Feb 3;243(4891):641-3
references.pmid: 2464851
references.citation: Nat Mater. 2009 Sep;8(9):736-41
references.pmid: 19684584
references.citation: J Neurosci Methods. 2003 Jul 15;127(1):75-84
references.pmid: 12865150
references.citation: Crit Rev Biotechnol. 2006 Oct-Dec;26(4):237-59
references.pmid: 17095434
references.citation: Genome Res. 2003 Oct;13(10):2348-52
references.pmid: 12975313
references.citation: Angew Chem Int Ed Engl. 2012 Nov 5;51(45):11224-40
references.pmid: 23111955
references.citation: Philos Trans A Math Phys Eng Sci. 2004 May 15;362(1818):923-35
references.pmid: 15306477
references.citation: Nat Biotechnol. 2008 Apr;26(4):417-26
references.pmid: 18392027
references.citation: Anal Biochem. 2008 Feb 1;373(1):141-6
references.pmid: 17868635
references.citation: Nat Med. 2003 Jan;9(1):140-5
references.pmid: 12514728