logging in or signing up presentation kristin Perrin Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINTLite Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: (To copy code, click on the text box) Embed: URL: Thumbnail: WordPress Embed Customize Embed The presentation is successfully added In Your Favorites. Views: 73 Category: Education License: All Rights Reserved Like it (0) Dislike it (0) Added: January 08, 2008 This Presentation is Public Favorites: 0 Presentation Description No description available. Comments Posting comment... Premium member Presentation Transcript Giant Unilamellar Vesicles: Giant Unilamellar Vesicles Kristin Ackermann Advisor: Dr. Steve Wrenn DREAM 2005Overview: Overview Giant unilamellar vesicles (GUVs) The importance of GUVs Preparation Methods Glass bead method Platinum wire method Cholesterol vesicles and domainsBackground: Background Vesicles are spheres consisting of a lipid bilayer and a water core GUVs are vesicles that are tens of micrometers in size http://academic.brooklyn.cuny.edu/biology/bio4fv/page/phosphb.htm GUVs have great potential in membrane researchObjectives: Objectives To describe step-by-step detailed procedures for making GUVs by both glass bead and platinum wire methods. To determine which method is more suitable for our applications in the lab. Experimental Methods Glass Bead Method: Experimental Methods Glass Bead Method Pyrex® Glass beads are added to cover the bottom of a 20 mL glass vial. A lipid mixture dissolved in chloroform is added to the vial The solution is dried under nitrogen A buffer is added and the vessel is heated at 70°C for three hoursResults So Far Glass Bead Method: Results So Far Glass Bead MethodExperimental Methods Platinum Wire Method: Experimental Methods Platinum Wire Method 50 uL of a lipid mixture is added to wire The wire is dried under vacuum for 30 minutes Buffer is added Voltage is then applied to the wire starting at 50mV 10 minutes: increase 5mV every minute 2 hours: increase 12 mV every 10 minutesResults So Far Platinum Wire Method: Results So Far Platinum Wire MethodWhat’s Next?: What’s Next? Run more experiments using the platinum wire method http://www.jlr.org/content/vol39/issue5/cover.shtml Use the atomic force microscope to measure the size of cholesterol domains in vesicles References: References Dimitrov, DS et al, FEBS 1980. Oct 1984, 176, 2, 398-400 H.H. Hub et al. FEBS Letters. Apr 1982, 140, 2, 254-256 Rodriguez, Nicolas et al. Coll & Surf B. 2005, 42, 125-130 Staneva, Galya et al. Chem & Phys of Lipids. 2004,129,53-62Acknowledgements : Acknowledgements Dr. Steve Wrenn Mona, Andrew and AngelaQuestions?: Questions? You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.
presentation kristin Perrin Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINTLite Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: (To copy code, click on the text box) Embed: URL: Thumbnail: WordPress Embed Customize Embed The presentation is successfully added In Your Favorites. Views: 73 Category: Education License: All Rights Reserved Like it (0) Dislike it (0) Added: January 08, 2008 This Presentation is Public Favorites: 0 Presentation Description No description available. Comments Posting comment... Premium member Presentation Transcript Giant Unilamellar Vesicles: Giant Unilamellar Vesicles Kristin Ackermann Advisor: Dr. Steve Wrenn DREAM 2005Overview: Overview Giant unilamellar vesicles (GUVs) The importance of GUVs Preparation Methods Glass bead method Platinum wire method Cholesterol vesicles and domainsBackground: Background Vesicles are spheres consisting of a lipid bilayer and a water core GUVs are vesicles that are tens of micrometers in size http://academic.brooklyn.cuny.edu/biology/bio4fv/page/phosphb.htm GUVs have great potential in membrane researchObjectives: Objectives To describe step-by-step detailed procedures for making GUVs by both glass bead and platinum wire methods. To determine which method is more suitable for our applications in the lab. Experimental Methods Glass Bead Method: Experimental Methods Glass Bead Method Pyrex® Glass beads are added to cover the bottom of a 20 mL glass vial. A lipid mixture dissolved in chloroform is added to the vial The solution is dried under nitrogen A buffer is added and the vessel is heated at 70°C for three hoursResults So Far Glass Bead Method: Results So Far Glass Bead MethodExperimental Methods Platinum Wire Method: Experimental Methods Platinum Wire Method 50 uL of a lipid mixture is added to wire The wire is dried under vacuum for 30 minutes Buffer is added Voltage is then applied to the wire starting at 50mV 10 minutes: increase 5mV every minute 2 hours: increase 12 mV every 10 minutesResults So Far Platinum Wire Method: Results So Far Platinum Wire MethodWhat’s Next?: What’s Next? Run more experiments using the platinum wire method http://www.jlr.org/content/vol39/issue5/cover.shtml Use the atomic force microscope to measure the size of cholesterol domains in vesicles References: References Dimitrov, DS et al, FEBS 1980. Oct 1984, 176, 2, 398-400 H.H. Hub et al. FEBS Letters. Apr 1982, 140, 2, 254-256 Rodriguez, Nicolas et al. Coll & Surf B. 2005, 42, 125-130 Staneva, Galya et al. Chem & Phys of Lipids. 2004,129,53-62Acknowledgements : Acknowledgements Dr. Steve Wrenn Mona, Andrew and AngelaQuestions?: Questions?