Development and fabrication of nanoporous silicon-based bioreactors within a microfluidic chip.
Multi-scale lithography and cryogenic deep reactive ion etching techniques were used to create ensembles of nanoporous, picolitre volume, reaction vessels within a microfluidic system. The fabrication of these vessels is described and how this process can be used to tailor vessel porosity by controlling the width of slits that constitute the vessel pores is demonstrated. Control of pore size allows the containment of nucleic acids and enzymes that are the foundation of biochemical reaction systems, while allowing smaller reaction constituents to traverse the container membrane and continuously supply the reaction. In this work, a 5.4 kb DNA plasmid was retained within the reaction vessels and labeled under microfluidic control with ethidium bromide as an initial proof-of-principle. Subsequently, a coupled enzyme reaction, in which glucose oxidase (GOX) and horseradish peroxidase (HRP) were contained and fed with a substrate solution of glucose and Amplex Red to produce fluorescent resorufin, was carried out under microfluidic control and monitored using fluorescent microscopy. The fabrication techniques presented are broadly applicable and can be adapted to produce devices in which a variety of high aspect ratio, nanoporous silicon structures can be integrated within a microfluidic network. The devices shown here are amenable to being scaled in number and organized to implement more complex reaction systems for applications in sensing and actuation as well as fundamental studies of biological reaction systems.
Retterer
Scott T
ST
Oak Ridge National Laboratory, Oak Ridge, TN, USA. rettererst@ornl.gov
Siuti
Piro
P
Choi
Chang-Kyoung
CK
Thomas
Darrell K
DK
Doktycz
Mitchel J
MJ
D019149:Bioreactors; D015374:Biosensing Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure*; D036103:Nanotechnology / Q000295:instrumentation*; D016062:Porosity; D012825:Silicon / Q000737:chemistry*
D012825:Silicon
Nat Rev Mol Cell Biol. 2009 Jun;10(6):410-22
19461664
J Phys Chem B. 2005 Feb 3;109(4):1609-17
16851132
Nanotechnology. 2008 Oct 15;19(41):
21394229
Trends Biotechnol. 2008 Jan;26(1):14-20
18037179
Nature. 2002 Nov 14;420(6912):224-30
12432407
Trends Biochem Sci. 2002 Jan;27(1):27-33
11796221
Genetica. 1999;106(1-2):49-55
10710709
Carbon N Y. 2008;46(11):1378-1383
19448842
J Membr Biol. 2003 Jan 15;191(2):87-97
12533776
Biotechnol Bioeng. 2005 Jan 5;89(1):1-8
15580587
Biochem Biophys Res Commun. 2007 Nov 9;363(1):12-7
17850764
Adv Drug Deliv Rev. 2004 Sep 22;56(11):1661-73
15350295
Anal Chem. 2009 Jun 15;81(12):4922-8
19441820
Trends Biotechnol. 2002 Mar;20(3):123-8
11841864
Angew Chem Int Ed Engl. 2006 Nov 13;45(44):7336-56
17086584
Proc Natl Acad Sci U S A. 2004 Dec 21;101(51):17669-74
15591347
Biomol Eng. 2001 Oct 31;18(4):143-77
11576871
Anal Chem. 2005 Mar 15;77(6):1539-44
15762555
Proc Natl Acad Sci U S A. 2008 Dec 23;105(51):20269-73
19074280
Cell. 2008 Oct 17;135(2):216-26
18957198
ACS Nano. 2008 Feb;2(2):247-54
19206624
Adv Drug Deliv Rev. 2004 Feb 10;56(2):211-29
14741117
Lab Chip. 2002 Nov;2(4):197-202
15100810
Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17290-5
17090676
Mol Syst Biol. 2007;3:125
17625513