LMP Seminar: Microfluidic tools for membrane biophysics and bottom-up synthetic biology
LMP Seminar
- Datum: 02.12.2019
- Uhrzeit: 14:00 - 15:30
- Vortragende(r): Dr. Tom Robinson
- Max Planck Institute of Colloids and Interfaces, Potsdam
- Ort: Max-Planck-Institut für Dynamik und Selbstorganisation (MPIDS)
- Raum: Riemannraum 1.40
- Gastgeber: MPIDS / LMP
- Kontakt: jaime.agudo@ds.mpg.de
Model membranes, and in particular, giant unilamellar vesicles (GUVs) are powerful tools for investigating the properties and functions of real cell membranes in a controlled manner in vitro. Here we present how microfluidic systems can be used to improve this field by aiding in the creation, manipulation, and analysis of GUVs (Robinson, Advanced Biosystems, 2019). After introducing the general principles of how micro-fabricated features can isolate defined numbers of these vesicles at specific spatial locations (Robinson et al. Biomicrofluidics 2013), we discuss how droplet-based microfluidics is used to create GUVs with superior size and encapsulation control.
Next, we introduce applications in the field of membrane biophysics. These include being able to apply a variety of external forces (electric fields, shear stress, and mechanical compression) to vesicles exhibiting domains as models for lipid rafts in biomembranes (Robinson et al. Lab on a Chip 2014; Sturzenegger, Robinson et al. Soft Matter 2016; Robinson et al. ChemBioChem 2019). We also utilise microfluidic systems to investigate the effects of solution asymmetry on the stabilisation of lipid domains and discuss the consequences of our findings in the context of cell membrane organisation (Kubsch, Robinson et al. Biophys. J. 2016). A high-throughput device is presented which we use to investigate membrane pore transport (Yandrapalli & Robinson, Lab on a Chip, 2019) as well as to measure the mechanical properties of lipid membranes.
Finally, we discuss how the combination of giant lipid vesicles and microfluidic systems is used in the field of bottom-up synthetic biology to create artificial cells. This includes two systems which mimic cell membrane fusion - one based on charged lipids (Lira, Robinson et al. Biophys. J. 2019) and the other using SNARE-mimetics for targeted fusion events. GUVs exhibiting multiple membrane compartments are presented as synthetic analogues to the architecture found in eukaryotic cells. These are used create out-of-equilibrium systems displaying self-organisation properties and reaction-diffusion systems using reconstituted membrane proteins.
Kubsch, B., Robinson, T., Lipowsky, R., and Dimova, R. (2016). Solution Asymmetry and Salt Expand Fluid-Fluid Coexistence Regions of Charged Membranes. Biophys. J. 110, 2581–2584.
Sturzenegger, F., Robinson, T., Hess, D., and Dittrich, P. S. (2016). Membranes under shear stress: visualization of non-equilibrium domain patterns and domain fusion in a microfluidic device. Soft Matter 12, 5072–5076.
Lira, R. B., Robinson, T., Dimova, R., and Riske, K. A. (2018). Highly Efficient Protein-free Membrane Fusion: A Giant Vesicle Study. Biophys. J. 116, 79–91.
Robinson, T. (2019). Microfluidic Handling and Analysis of Giant Vesicles for Use as Artificial Cells: A Review. Adv. Biosyst., 1800318.
Robinson, T., and Dittrich, P. S. (2019). Observations of membrane domain reorganization in mechanically compressed artificial cells. ChemBioChem, cbic.201900167.
Yandrapalli, N., and Robinson, T. (2019). Ultra-high capacity microfluidic trapping of giant vesicles for high-throughput membrane studies. Lab Chip 19, 626–633.
Robinson, T., Kuhn, P., Eyer, K., and Dittrich, P. S. (2013). Microfluidic trapping of giant unilamellar vesicles to study transport through a membrane pore. Biomicrofluidics 7, 44105.
Robinson, T., Verboket, P. E., Eyer, K., and Dittrich, P. S. (2014). Controllable electrofusion of lipid vesicles: initiation and analysis of reactions within biomimetic containers. Lab Chip, 2852–2859.
Next, we introduce applications in the field of membrane biophysics. These include being able to apply a variety of external forces (electric fields, shear stress, and mechanical compression) to vesicles exhibiting domains as models for lipid rafts in biomembranes (Robinson et al. Lab on a Chip 2014; Sturzenegger, Robinson et al. Soft Matter 2016; Robinson et al. ChemBioChem 2019). We also utilise microfluidic systems to investigate the effects of solution asymmetry on the stabilisation of lipid domains and discuss the consequences of our findings in the context of cell membrane organisation (Kubsch, Robinson et al. Biophys. J. 2016). A high-throughput device is presented which we use to investigate membrane pore transport (Yandrapalli & Robinson, Lab on a Chip, 2019) as well as to measure the mechanical properties of lipid membranes.
Finally, we discuss how the combination of giant lipid vesicles and microfluidic systems is used in the field of bottom-up synthetic biology to create artificial cells. This includes two systems which mimic cell membrane fusion - one based on charged lipids (Lira, Robinson et al. Biophys. J. 2019) and the other using SNARE-mimetics for targeted fusion events. GUVs exhibiting multiple membrane compartments are presented as synthetic analogues to the architecture found in eukaryotic cells. These are used create out-of-equilibrium systems displaying self-organisation properties and reaction-diffusion systems using reconstituted membrane proteins.
Kubsch, B., Robinson, T., Lipowsky, R., and Dimova, R. (2016). Solution Asymmetry and Salt Expand Fluid-Fluid Coexistence Regions of Charged Membranes. Biophys. J. 110, 2581–2584.
Sturzenegger, F., Robinson, T., Hess, D., and Dittrich, P. S. (2016). Membranes under shear stress: visualization of non-equilibrium domain patterns and domain fusion in a microfluidic device. Soft Matter 12, 5072–5076.
Lira, R. B., Robinson, T., Dimova, R., and Riske, K. A. (2018). Highly Efficient Protein-free Membrane Fusion: A Giant Vesicle Study. Biophys. J. 116, 79–91.
Robinson, T. (2019). Microfluidic Handling and Analysis of Giant Vesicles for Use as Artificial Cells: A Review. Adv. Biosyst., 1800318.
Robinson, T., and Dittrich, P. S. (2019). Observations of membrane domain reorganization in mechanically compressed artificial cells. ChemBioChem, cbic.201900167.
Yandrapalli, N., and Robinson, T. (2019). Ultra-high capacity microfluidic trapping of giant vesicles for high-throughput membrane studies. Lab Chip 19, 626–633.
Robinson, T., Kuhn, P., Eyer, K., and Dittrich, P. S. (2013). Microfluidic trapping of giant unilamellar vesicles to study transport through a membrane pore. Biomicrofluidics 7, 44105.
Robinson, T., Verboket, P. E., Eyer, K., and Dittrich, P. S. (2014). Controllable electrofusion of lipid vesicles: initiation and analysis of reactions within biomimetic containers. Lab Chip, 2852–2859.