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JoVE Journal Bioengineering
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Bioengineering

Microfluidic pikoliter bioreaktor for Microbial Encellede Analyse: Fabrikasjon, System Setup, og drift

Published: December 6, 2013 doi: 10.3791/50560
Automatic Translation
1, 1, 1, 1, 1, 1
1Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Juelich GmbH

Materials

Name Company Catalog Number Comments
Silicon wafer 100 mm diameter, P/BOR <100> Si-MAT, Silicon Materials, Germany
Photoresist SU-8 2000.5 Micro Resist Technology GmbH, Germany
Photoresist SU-8 2010 Micro Resist Technology GmbH, Germany
SU-8 Developer mr DEV- 600 Micro Resist Technology GmbH, Germany
Polydimethylsiloxane (PDMS) Sylgard 184 Silicone Elastomer Kit Dow Corning; Farnell GmbH, Germany
Dispensing needles Precision Tips  27 G; ID = 0.2 mm, OD = 0.42 mm Nordson EFD Deutschland, Germany
Glass plates D263 T eco, 30 mm x 25 mm x 0.17 mm Schott AG, Germany
Hole puncher AKA 5130-B-90 Harris Uni-Core
Tubing Tygon S-54-HL, ID = 0.25 mm, OD = 0.76 mm Saint Gobain; VWR International GmbH, Germany
Disposable Syringes – Omnifix Spritzen BRAUN Omnifix 40 Duo, 1 ml B. Braun Melsungen AG, Germany 552-183143
Syringes, 1 ml sterile glass syringes INNOVATIVE LABOR SYSTEME GMBH (ILS), Germany
Chemicals
(NH4)2SO4 Carl Roth GmbH + Co. KG, Germany
Urea Carl Roth GmbH + Co. KG, Germany
KH2PO4 Carl Roth GmbH + Co. KG, Germany
K2HPO4 Carl Roth GmbH + Co. KG, Germany
MgSO4•7H2O Carl Roth GmbH + Co. KG, Germany
MOPS Carl Roth GmbH + Co. KG, Germany
FeSO4•7H2O Carl Roth GmbH + Co. KG, Germany
MnSO4•H2O Carl Roth GmbH + Co. KG, Germany
ZnSO4•7H2O Carl Roth GmbH + Co. KG, Germany
CuSO4 Carl Roth GmbH + Co. KG, Germany
NiCl2•6H2O Carl Roth GmbH + Co. KG, Germany
CaCl2 Carl Roth GmbH + Co. KG, Germany
Biotin Carl Roth GmbH + Co. KG, Germany
Procatechuic acid Carl Roth GmbH + Co. KG, Germany
Glucose-Monohydrate Carl Roth GmbH + Co. KG, Germany
BHI Becton, Dickinson
Cells
Corynebacterium glutamicum ATTC 13032 DSMZ; Leibniz-Institut DSMZ - Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Germany
Escherichia coli MG 1655 DSMZ; Leibniz-Institut DSMZ - Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Germany
Equipment
Wafer Cleaner SSEC 3300 Solid State Equipment LLC
Spin Coater SPIN150 -NPP SPS Europe B.V.
Mask Aligner MA-6 Karl Suess
Hot Plate HP30A - 2 Torrey Pines Scientific
Laboratory oven Memmert UN 200 Memmert
Plasma Cleaner FEMTO Diener Electronics, Germany
neMESYS syringe pumps Cetoni GmbH, Germany
Magnetic stirrer CB 162 Stuart VWR 442-0304
Microscope Nikon Eclipse Ti Nikon Microscopy
Microscope incubuator Pecon GmbH, Germany
Centrifuge minispin plus “black line” Eppendorf 9776501
Photometer BioPhotometer plus  Eppendorf 6132000008
Shake flask shaker/incubator 3031 GFL - Gesellschaft für Labortechnik mbH, Germany
Profilometer, Dektak 150 Stylus Profiler Veeco

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References

  1. Muzzey, D., van Oudenaarden, A. Quantitative Time-Lapse Fluorescence Microscopy in Single Cells. Ann. Rev. Cell Dev. Biol. 25, 301-327 (2009).
  2. Wei, P., et al. Bacterial virulence proteins as tools to rewire kinase pathways in yeast and immune cells. Nature. 488, 384-388 (2012).
  3. Park, S., et al. Influence of topology on bacterial social interaction. Proc. Natl. Acad. Sci. U.S.A. 100, 13910-13915 (1073).
  4. Lee, P. J., Helman, N. C., Lim, W. A., Hung, P. J. A microfluidic system for dynamic yeast cell imaging. BioTechniques. 44, 91-95 (2008).
  5. Kobel, S. A., et al. Automated analysis of single stem cells in microfluidic traps. Lab Chip. 12, 2843-2849 (2012).
  6. Carlo, D. D., Wu, L. Y., Lee, L. P. Dynamic single cell culture array. Lab Chip. 6, 1445-1449 (2006).
  7. Locke, J. C. W., Young, J. W., Fontes, M., Jimenez, M. J. H., Elowitz, M. B. Stochastic Pulse Regulation in Bacterial Stress Response. Science. 334, 366-369 (2011).
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  9. Fritzsch, F. S. O., et al. Picoliter nDEP traps enable time-resolved contactless single bacterial cell analysis in controlled microenvironments. Lab Chip. 13, 397-408 (2013).
  10. Groisman, A., et al. A microfluidic chemostat for experiments with bacterial and yeast cells. Nat. Methods. 2, 685-689 (2005).
  11. Mather, W., Mondragon-Palomino, O., Danino, T., Hasty, J., Tsimring, L. S. Streaming Instability in Growing Cell Populations. Phys. Rev. Lett. 104, (2010).
  12. Wang, P., et al. Robust growth of Escherichia coli. Curr. Biol. 20, 1099-1103 (2010).
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  14. Walden, M., Elf, J. Studying transcriptional interactions in single cells at sufficient resolution. Curr. Opin. Biotech. 22, 81-86 (2011).
  15. Keymer, J. E., Galajda, P., Lambert, G., Liao, D., Austin, R. H. Computation of mutual fitness by competing bacteria. Proc. Natl. Acad. Sci. U.S.A. 105, 20269-20273 (2008).
  16. Keymer, J. E., Galajda, P., Muldoon, C., Park, S., Austin, R. H. Bacterial metapopulations in nanofabricated landscapes. Proc. Natl. Acad. Sci. U.S.A. 103, 17290-17295 (2006).
  17. Grunberger, A., et al. Beyond growth rate 0.6: Corynebacterium glutamicum cultivated in highly diluted environments. Biotechnol. Bioeng. 110, 220-228 (2013).
  18. Binder, S., et al. A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level. Genome Biol. 13, (2012).
  19. Mustafi, N., Grunberger, A., Kohlheyer, D., Bott, M., Frunzke, J. The development and application of a single-cell biosensor for the detection of L-methionine and branched-chain amino acids. Metab. Eng. 14, 449-457 (2012).
  20. Huber, R., et al. Robo-Lector - a novel platform for automated high-throughput cultivations in microtiter plates with high information content. Microb. Cell Fact. 8, (2009).
  21. Keilhauer, C., Eggeling, L., Sahm, H. Isoleucine synthesis in Corynebacterium glutamicum: molecular analysis of the ilvB-ilvN-ilvC operon. J. Bacteriol. 175, 5595-5603 (1993).
  22. Frunzke, J., Bramkamp, M., Schweitzer, J. E., Bott, M. Population Heterogeneity in Corynebacterium glutamicum ATCC 13032 caused by prophage CGP3. J. Bacteriol. 190, 5111-5119 (2008).
Microfluidic pikoliter bioreaktor for Microbial Encellede Analyse: Fabrikasjon, System Setup, og drift
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Cite this Article

Gruenberger, A., Probst, C., Heyer,… More

Gruenberger, A., Probst, C., Heyer, A., Wiechert, W., Frunzke, J., Kohlheyer, D. Microfluidic Picoliter Bioreactor for Microbial Single-cell Analysis: Fabrication, System Setup, and Operation. J. Vis. Exp. (82), e50560, doi:10.3791/50560 (2013).

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