Abstract
We introduce a micro-scale bioreactor for automated culture and density analysis of microorganisms. The microbioreactor is powered by digital microfluidics (DMF) and because it is used with bacteria, algae and yeast, we call it the BAY microbioreactor. Previous miniaturized bioreactors have relied on microchannels which often require valves, mixers and complex optical systems. In contrast, the BAY microbioreactor is capable of culturing microorganisms in distinct droplets on a format compatible with conventional bench-top analyzers without the use of valves, mixers or pumps. Bacteria, algae and yeast were grown for up to 5 days with automated semi-continuous mixing and temperature control. Cell densities were determined by measuring absorbances through transparent regions of the devices, and growth profiles were shown to be comparable to those generated in conventional, macro-scale systems. Cell growth and density measurements were integrated in the microbioreactor with a fluorescent viability assay and transformation of bacteria with a fluorescent reporter gene. These results suggest that DMF may be a useful new tool in automated culture and analysis of microorganisms for a wide range of applications.
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M. Abdelgawad, M.W. Watson, A.R. Wheeler, Lab Chip 9, 1046 (2009)
M. Abdelgawad, A.R. Wheeler, Adv. Mat 21, 920 (2009)
M. Abdelgawad, S.L. Freire, H. Yang, A.R. Wheeler, Lab Chip 8, 672 (2008)
S.N. Bailey, R.Z. Wu, D.M. Sabatini, Drug Discov Today 7, S113 (2002)
S.N. Bailey, D.M. Sabatini, B.R. Stockwell, Proc Natl Acad Sci USA 101, 16144 (2004)
F.K. Balagadde, L. You, C.L. Hansen, F.H. Arnold, S.R. Quake, Science 309, 137 (2005)
I. Barbulovic-Nad, S.H. Au, A.R. Wheeler, Lab Chip 10, 1536 (2010)
I. Barbulovic-Nad, H. Yang, P.S. Park, A.R. Wheeler, Lab Chip 8, 519 (2008)
G.P.L. Cereghino, J.M. Cregg, Curr Opin Biotechnol 10, 422 (1999)
Y.H. Chang, G.B. Lee, F.C. Huang, Y.Y. Chen, J.L. Lin, Biomed Microdevices 8, 215 (2006)
D. Chatterjee, A.J. Ytterberg, S.U. Son, J.A. Loo, R.L. Garrell, Anal Chem 82, 2095 (2010)
Y. Chisti, Trends Biotechnol 26, 126 (2008)
A.L. Demain, J.L. Adrio, Mol Biotechnol 38, 41 (2008)
S. Faley, K. Seale, J. Hughey, D. K. Schaffer, S. VanCompernolle, B. McKinney, F. Baudenbacher, D. Unutmaz, J. P. Wikswo, Lab Chip. 8, 1700 (2008)
S. L. Faley, M. Copland, D. Wlodkowic, W. Kolch, K. T. Seale, J. P. Wikswo, J. M. Cooper, Lab Chip. 9, 2659 (2009)
G.B. Fogel, C.F. Brunk, Anal Biochem 260, 80 (1998)
J. Fowler, M. Hyejin, K. Chang-Jin, in Micro Electro Mechanical Systems, 2002. The Fifteenth IEEE International Conference on, p. 97–100 (2002)
G. Giaever, A.M. Chu, L. Ni, C. Connelly, L. Riles, S. Veronneau, S. Dow, A. Lucau-Danila, K. Anderson, B. Andre, A.P. Arkin, A. Astromoff, M. El Bakkoury, R. Bangham, R. Benito, S. Brachat, S. Campanaro, M. Curtiss, K. Davis, A. Deutschbauer, K.-D. Entian, P. Flaherty, F. Foury, D.J. Garfinkel, M. Gerstein, D. Gotte, U. Guldener, J.H. Hegemann, S. Hempel, Z. Herman, D.F. Jaramillo, D.E. Kelly, S.L. Kelly, P. Kotter, D. LaBonte, D.C. Lamb, N. Lan, H. Liang, H. Liao, L. Liu, C. Luo, M. Lussier, R. Mao, P. Menard, S.L. Ooi, J.L. Revuelta, C.J. Roberts, M. Rose, P. Ross-Macdonald, B. Scherens, G. Schimmack, B. Shafer, D.D. Shoemaker, S. Sookhai-Mahadeo, R.K. Storms, J.N. Strathern, G. Valle, M. Voet, G. Volckaert, C-y Wang, T.R. Ward, J. Wilhelmy, E.A. Winzeler, Y. Yang, G. Yen, E. Youngman, K. Yu, H. Bussey, J.D. Boeke, M. Snyder, P. Philippsen, R.W. Davis, M. Johnston, Nature 418, 387 (2002)
A. Groisman, C. Lobo, H. Cho, J.K. Campbell, Y.S. Dufour, A.M. Stevens, A. Levchenko, Nat Methods 2, 685 (2005)
A. Humphrey, Biotechnol Prog 14, 3 (2008)
M.J. Jebrail, A.R. Wheeler, Anal Chem 81, 330 (2009)
M. J. Jebrail, V. N. Luk, S. C. C. Shih, R. Fobel, A. H. Ng, H. Yang, S. L. Freire, A. R. Wheeler, J Vis Exp. (2009). doi:10.3791/1603
J.S. Kee, D.P. Poenar, P. Neuzil, L. Yobas, Sens and Act. B 134, 532 (2008)
E.V. Koonin, M.Y. Galperin, Curr Opin Genet Dev 7, 757 (1997)
B.H. Lapizco-Encinas, B.A. Simmons, E.B. Cummings, Y. Fintschenko, Anal Chem 76, 1571 (2004)
H.L. Lee, P. Boccazzi, R.J. Ram, A.J. Sinskey, Lab Chip 6, 1229 (2006)
Z. Liang, N. Chiem, G. Ocvirk, T. Tang, K. Fluri, D.J. Harrison, Anal Chem 68, 1040 (1996)
A. Llobera, S. Demming, R. Wilke, S. Buttgenbach, Lab Chip 7, 1560 (2007)
H.W. Lu, F. Bottausci, J.D. Fowler, A.L. Bertozzi, C. Meinhart, C.J. Kim, Lab Chip 8, 456 (2008)
V.N. Luk, G. Mo, A.R. Wheeler, Langmuir 24, 6382 (2008)
V.N. Luk, A.R. Wheeler, Anal Chem 81, 4524 (2009)
M. Mandel, A. Higa, J Mol Biol 53, 159 (1970)
E.M. Miller, A.R. Wheeler, Anal Chem 80, 1614 (2008)
D.J.S. Montagnes, D.J. Franklin, Limnol. Oceangr. 46, 2008 (2001)
H. Moon, A.R. Wheeler, R.L. Garrell, J.A. Loo, C.J. Kim, Lab Chip 6, 1213 (2006)
N. A. Mousa, M. J. Jebrail, H. Yang, M. Abdelgawad, P. Metalnikov, J. Chen, A. R. Wheeler, R. F. Casper, Sci Transl Med. 1, 1ra2 (2009)
P. Paik, V.K. Pamula, R.B. Fair, Lab Chip 3, 253 (2003a)
P. Paik, V.K. Pamula, M.G. Pollack, R.B. Fair, Lab Chip 3, 28 (2003b)
X.Y. Peng, P.C.H. Li, Anal Chem 76, 5282 (2004)
J. Piškur, R.B. Langkjær, Mol Microbiol 53, 381 (2004)
D.A. Ratkowsky, J. Olley, T.A. McMeekin, A. Ball, J Bacteriol 149, 1 (1982)
J. Ryley, O.M. Pereira-Smith, Yeast 23, 1065 (2006)
G.J. Shah, A.T. Ohta, E.P. Chiou, M.C. Wu, C.J. Kim, Lab Chip 9, 1732 (2009)
R. Sista, Z. Hua, P. Thwar, A. Sudarsan, V. Srinivasan, A. Eckhardt, M. Pollack, V. Pamula, Lab Chip 8, 2091 (2008a)
R.S. Sista, A.E. Eckhardt, V. Srinivasan, M.G. Pollack, S. Palanki, V.K. Pamula, Lab Chip 8, 2188 (2008b)
S. Son, R. Garrell, Lab Chip 9, 2398 (2009)
V. Srinivasan, V.K. Pamula, R.B. Fair, Anal Chim Acta 507, 145 (2004a)
V. Srinivasan, V.K. Pamula, R.B. Fair, Lab Chip 4, 310 (2004b)
J.R. Swartz, Curr Opin Biotechnol 12, 195 (2001)
M.D. Vahey, J. Voldman, Anal Chem 80, 3135 (2008)
E. Verpoorte, A. Manz, H. Ludi, A.E. Bruno, F. Maystre, B. Krattiger, H.M. Widmer, B.H. van der Schoot, N.F. de Roojj, Sens and Act B. 6, 66 (1992)
I. Walther, B.v d Schootb, M. Boillatb, A. Cogoli, Enzyme Microb Technol 27, 778 (2000)
M.W. Watson, M.J. Jebrail, A.R. Wheeler, Anal Chem 82, 6680 (2010)
A.R. Wheeler, H. Moon, C.A. Bird, R.R.O. Loo, C.J. Kim, J.A. Loo, R.L. Garrell, Anal Chem 77, 534 (2005)
A.R. Wheeler, Science 322, 539 (2008)
E. Yu, F. Zendejas, P. Lane, S. Gaucher, B. Simmons, T. Lane, Jour App. Phycol. (2009). doi: 10.1007/s10811-008-9400-y
Z. Zhang, P. Boccazzi, H.G. Choi, G. Perozziello, A.J. Sinskey, K.F. Jensen, Lab Chip 6, 906 (2006)
J. Ziauddin, D.M. Sabatini, Nature 411, 107 (2001)
Acknowledgements
We thank Prof. Kevin Truong (Institute of Biomaterials and Biomedical Engineering, IBBME, University of Toronto) for generously donating E. coli and plasmid DNA, Prof. Igor Stagjlar (Department of Medical Genetics and Microbiology, University of Toronto) for generously donating S. cerevisiae and Prof. William Ryu (Department of Physics, University of Toronto) for the use of the cold room. We also thank Evan Mills (IBBME, University of Toronto) and Dawn Edmonds (Department of Medical Genetics and Microbiology, University of Toronto) for their assistance with bacterial transformation and yeast culture. We thank Kamlesh D. Patel and Pam Lane (Sandia National Laboratories, Livermore, CA) for discussion and assistance with algae culture. We thank the Canadian Institutes of Health Research (CIHR) for financial support. SHA and SCCS thank NSERC (Natural Sciences and Engineering Research Council) for graduate fellowships, and ARW thanks CRC for a Canada Research Chair.
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Sam H. Au and Steve C. C. Shih contributed equally
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Au, S.H., Shih, S.C.C. & Wheeler, A.R. Integrated microbioreactor for culture and analysis of bacteria, algae and yeast. Biomed Microdevices 13, 41–50 (2011). https://doi.org/10.1007/s10544-010-9469-3
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DOI: https://doi.org/10.1007/s10544-010-9469-3