Summary
Quantitative studies of chemotactic signaling require experimental techniques that can expose single cells to chemical stimuli with high resolution in both space and time. Recently, we have introduced the method of flow photolysis (Anal. Chem. 79:3940–3944, 2007), which combines microfluidic techniques with the photochemical release of caged compounds. This method allows us to tailor chemical stimuli on the length scale of individual cells with subsecond temporal resolution. In this chapter, we provide a detailed protocol for the setup of flow photolysis experiments and exemplify this versatile approach by initiating membrane translocation of fluorescent fusion proteins in chemotactic Dictyostelium discoideum cells.
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References
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Acknowledgments
The flow photolysis approach was developed at the Max Planck Institute for Dynamics and Self-Organization in Göttingen, Germany, together with Danica Wyatt and Eberhard Bodenschatz. The initial ideas were designed in collaboration with Wouter-Jan Rappel from UCSD. We thank Loling Song and Sharvari Nadkarni for valuable contributions to our soft lithography protocol. Inspiring discussions with Albert Bae, Gabriel Amselem, Christian Westendorf, and William Loomis are acknowledged.
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Beta, C. (2009). Spatiotemporal Stimulation of Single Cells Using Flow Photolysis. In: Jin, T., Hereld, D. (eds) Chemotaxis. Methods in Molecular Biology™, vol 571. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-198-1_22
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DOI: https://doi.org/10.1007/978-1-60761-198-1_22
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