Development of Giant Unilamellar Vesicles as long as the Study of Crowded Protein Environments David D. Gooray, S in addition to eep Dhall, in addition to Victor G. J. Rodgers Bioengineering Department, University of Cali as long as nia, Riverside Conclusion & Future Work

Development of Giant Unilamellar Vesicles as long as the Study of Crowded Protein Environments David D. Gooray, S in addition to eep Dhall, in addition to Victor G. J. Rodgers Bioengineering Department, University of Cali as long as nia, Riverside Conclusion & Future Work www.phwiki.com

Development of Giant Unilamellar Vesicles as long as the Study of Crowded Protein Environments David D. Gooray, S in addition to eep Dhall, in addition to Victor G. J. Rodgers Bioengineering Department, University of Cali as long as nia, Riverside Conclusion & Future Work

Lucke, Mark, News Director has reference to this Academic Journal, PHwiki organized this Journal Development of Giant Unilamellar Vesicles as long as the Study of Crowded Protein Environments David D. Gooray, S in addition to eep Dhall, in addition to Victor G. J. Rodgers Bioengineering Department, University of Cali as long as nia, Riverside Giant unilamellar vesicles (GUVs) are supramolecular structures consisting of amphiphiles that range in sizes from 10-100 µm.[1] They provide an easy method to: (1) observe environment as long as in-vitro studies of compartmentalized reactions in addition to (2) model particular cell behavior. By studying the effects of crowded protein environments within GUVs, we will achieve a greater underst in addition to ing of processes in addition to properties such as mitochondrial swelling in addition to osmotic pressure. Phosphatidylcholine lipids are the only lipids that can as long as m GUVs under electro as long as mation.[2] GUVs with a diameter ranging between 50-100 µm will withst in addition to micromanipulation techniques in addition to allow us to study the effects of crowded protein environments. We prepared GUVs using the 3 types of phosphatidylcholine lipids; 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), L–phosphatidylcholine (Egg PC) in addition to 1-2-dioleoyl-sn-glyeo-3-phosphocholine(DPC). A square chamber was cut from teflon in addition to molded to fit the microscope slide. Two cylindrical platinum wires (Sigma-Aldrich) of 1.0 mm diameter were housed approximately parallel to each other with a 3.0 mm separation Each wire was 1.5 mm from transparent viewing slide All drilled holes in addition to platinum-teflon interfaces were sealed with silicone Conclusion & Future Work Results (Continued) Results Acknowledgements B2K Group, Dr. Vullev, Dr. Anvari, in addition to Dr. Park Jun Wang in addition to Bioengineering department. The GUVs selected as long as micromanipulation had an average size of 66.9 µm in addition to were observed using a light microscope with a phase contrast filter. Average diameter of GUVs were determined using ImageJ. The average diameters of the GUVs obtained are: POPC GUVs – 52.5 µm, EPC GUVs – 67.6 µm, in addition to DOPC GUVs – 70.4 µm We successfully setup the experiments as long as preparing GUVs in addition to obtained the required sizes. GUVs with individual distribution on the wire were dependent on the precision of lipid application on the wire. The future direction includes loading of the GUVs with proteins while surrounding their environments with a similar concentration of the protein. This is to reduce the osmotic pressure within the GUVs in addition to their respective loading characteristics. We will investigate better microinjection apparatus to enhance the precision of our piercings in addition to load delivery into the GUVs. Materials & MethodsYa Stock solutions of 1-palmitoyl-2-oleoyl-sn-glycero-3- phosphocholine (POPC), L–lysophosphatidylcholine (EPC) in addition to 1-2-dioleoyl-sn-glyeo-3-phosphocholine (DPC) were made. (Avanti Polar Lipids) Frequency: 10 Hz A-C Voltage Ramp: 0.3 V increased to 2.3 V in 15 min 2.3 V applied as long as 105 min [1]Luisi, P. in addition to Walde, P. Giant Vesicles. New York: John Wiley & Sons Ltd. 2000 [2]Angelova,M.I., S.Soleau, P.Melead,J.- Faucon, in addition to P.Bothorel. Preperation of giant vesicles by external ac electric fields: kinetics in addition to applications.Prog. Colloid Polym. Sci. 89:127-131(1992) Experimental Setup References Single GUV Experimental set up Size distribution of GUVs Introduction

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