Abstract
A three-point microfluidic system was developed and used to experimentally verify bacterial chemotaxis with known chemoeffectors. Using pneumatically-controlled micro-valves, the device was able to regulate microscale flows and created concentration gradients that allowed GFP-labelled Escherichia coli cells to interact with an environment that contained a chemoattractant and a chemorepellent. Having two separate possible paths (left and right) for the bacteria to move forward, this device also allowed for imaging processing based removal of noisy data, if adirectional bias was present. This device could be useful for quantitative analysis of chemotactic behaviors with minimal technical requirements, and could motivate the development of future devices based on this concept.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Whitesides, G.M. The origins and the future of microfluidics. Nature 442, 368–373 (2006).
Kim, M.J. & Breuer, K.S. A Selective Mixing in Microfluidic Systems Using Bacterial Chemotaxis. in ASME 277–282 (2005).
Kim, S., Kim, H.J. & Jeon, N.L. Biological applications of microfluidic gradient devices. Integr. Biol. 2, 584–603 (2010).
Weibel, D.B. & Whitesides, G.M. Applications of microfluidics in chemical biology. Curr. Opin. Chem. Biol. 10, 584–591 (2006).
Sia, S.K. & Whitesides, G.M. Microfluidic devices fabricated in poly (dimethylsiloxane) for biological studies. Electrophoresis 24, 3563–3576 (2003).
Weibel, D.B., DiLuzio, W.R. & Whitesides, G.M. Microfabrication meets microbiology. Nat. Rev. Microbiol. 5, 209–218 (2007).
Zare, R.N. & Kim, S. Microfluidic platforms for single- cell analysis. Annu. Rev. Biomed. Eng. 12, 187–201 (2010).
Li, J. & Lin, F. Microfluidic devices for studying chemotaxis and electrotaxis. Trends Cell Biol. 21, 489–497 (2011).
Meyvantsson, I. & Beebe, D.J. Cell culture models in microfluidic systems. Annu. Rev. Anal. Chem. 1, 423–449 (2008).
Ahmed, T., Shimizu, T.S. & Stocker, R. Bacterial chemotaxis in linear and nonlinear steady microfluidic gradients. Nano Lett. 10, 3379–3385 (2010).
Keenan, T.M. & Folch, A. Biomolecular gradients in cell culture systems. Lab Chip 8, 34–57 (2007).
Diao, J. et al. A three-channel microfluidic device for generating static linear gradients and its application to the quantitative analysis of bacterial chemotaxis. Lab Chip 6, 381–388 (2006).
Wadhams, G.H. & Armitage, J.P. Making sense of it all: bacterial chemotaxis. Nat. Rev. Mol. Cell Biol. 5, 1024–1037 (2004).
Eisenbach, M. Bacterial Chemotaxis. in eLS (John Wiley & Sons, Ltd, 2001).
Kim, M.J. & Breuer, K.S. Controlled mixing in microfluidic systems using bacterial chemotaxis. Anal. Chem. 79, 955–959 (2007).
Phuyal, K. & Kim, M.J. Mechanics of swimming of multi-body bacterial swarmers using non-labeled cell tracking algorithm. Phys. Fluids B 25, 011901 (2013).
Berg, H.C. Motile behavior of bacteria. Phys. Today 53, 24–30 (2000).
Darnton, N.C., Turner, L., Rojevsky, S. & Berg, H.C. Dynamics of bacterial swarming. Biophys. J. 98, 2082–2090 (2010).
Ford, R.M. & Harvey, R.W. Role of chemotaxis in the transport of bacteria through saturated porous media. Adv. Water Resour. 30, 1608–1617 (2007).
Paguirigan, A.L. & Beebe, D.J. Microfluidics meet cell biology: bridging the gap by validation and application of microscale techniques for cell biological assays. BioEssays 30, 811–821 (2008).
Jeon, N.L. et al. Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device. Nat. Biotechnol. 20, 826–830 (2002).
Chung, B.G. et al. A gradient-generating microfluidic device for cell biology. J. Vis. Exp. 7 (2007).
Jeon, N.L. et al. Generation of solution and surface gradients using microfluidic systems. Langmuir 16, 8311–8316 (2000).
Cheng, S.-Y. et al. A hydrogel-based microfluidic device for the studies of directed cell migration. Lab Chip 7, 763–769 (2007).
Haessler, U., Kalinin, Y., Swartz, M.A. & Wu, M. An agarose-based microfluidic platform with a gradient buffer for 3D chemotaxis studies. Biomed. Microdevices 11, 827–835 (2009).
Atencia, J. & Beebe, D.J. Controlled microfluidic interfaces. Nature 437, 648–655 (2005).
Beebe, D.J., Mensing, G.A. & Walker, G.M. Physics and applications of microfluidics in biology Annu. Rev. Biomed. Eng. 4, 261–286 (2002).
Mengeaud, V., Josserand, J. & Girault, H.H. Mixing processes in a zigzag microchannel: finite element simulations and optical study. Anal. Chem. 74, 4279–4286 (2002).
Studer, V. et al. Scaling properties of a low-actuation pressure microfluidic valve. J. Appl. Phys. 95, 393–398 (2003).
Kamholz, A.E., Weigl, B.H., Finlayson, B.A. & Yager, P. Quantitative analysis of molecular interaction in a microfluidic channel: the T-sensor. Anal. Chem. 71, 5340–5347 (1999).
Erickson, D. Towards numerical prototyping of labson- chip: modeling for integrated microfluidic devices. Microfluid. Nanofluid. 1, 301–318 (2005).
Berg, H.C. E. coli in Motion, (Springer, 2004).
Alon, U., Surette, M.G., Barkai, N. & Leibler, S. Robustness in bacterial chemotaxis. Nature 397, 168–171 (1999).
Adler, J. A method for measuring chemotaxis and use of the method to determine optimum conditions for chemotaxis by Escherichia coli. J. Gen. Microbiol. 74, 77–91 (1973).
Mao, H., Cremer, P.S. & Manson, M.D. A sensitive, versatile microfluidic assay for bacterial chemotaxis. PNAS 100, 5449–5454 (2003).
De Pina, K., Desjardin, V., Mandrand-Berthelot, M. -A., Giordano, G. & Wu, L.-F. Isolation and Characterization of thenikR Gene Encoding a Nickel-Responsive Regulator in Escherichia coli. J. Bacteriol. 181, 670–674 (1999).
Borrok, D., Borrok, M.J., Fein, J.B. & Kiessling, L.L. Link between chemotactic response to Ni2+ and its adsorption onto the Escherichia coli cell surface. Environ. Sci. Technol. 39, 5227–5233 (2005).
Macomber, L. & Hausinger, R.P. Mechanisms of nickel toxicity in microorganisms. Metallomics 3, 1153–1162 (2011).
Unger, M.A., Chou, H.-P., Thorsen, T., Scherer, A. & Quake, S.R. Monolithic microfabricated valves and pumps by multilayer soft lithography. Science 288, 113–116 (2000).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kim, H., Ali, J., Phuyal, K. et al. Investigation of bacterial chemotaxis using a simple three-point microfluidic system. BioChip J 9, 50–58 (2015). https://doi.org/10.1007/s13206-014-9107-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13206-014-9107-x