IGEM:IMPERIAL/2007/Projects/In-Veso/Presentations/29-08-07

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Vesicles

IC07 runner index.png
  1. Method of vesicle formation
  2. Emulsions, briefly
  3. Progress to date
  4. The Pore Dilemma
  5. The next steps

The Method

IC07 runner index 1.png

Stage 1 Stage 2 Stage 3
IC07 Suspension.png IC07 Emulsion.png IC07 Interface.png
Preparation of lipid-oil suspension Emulsification of vesicle contents Bi-layer formed through sedimentation

Emulsions

IC07 runner index 2.png

We are mixing oil and water - a very old problem...

  • The problem arises due to a high surface tension at the interface.
  • The creation of surfaces requires an energy input - work.
  • The lowest energy state is that with smallest interface surface area.

Surfactants are used to reduce surface tension.

IC07 surfacetension.png

A surfactant is any substance that reduces the surface tension between two immiscible liquids.

Art, not Science

IC07 runner index 2.png

The activity of surfactants depends on:

  • Oil composition
  • Salinity and osmolarity
  • pH, temperature, pressure
  • Head and tail characteristics
  • etc.
IC07 davincipoly.png

The process is hard to predict and reproduce.
Our understanding of the process is qualitative.
The physical laws are still unknown.



What We have Done

IC07 runner index 3.png

We think we encapsulated GFP in vesicles.

IC07 image120-red.jpg
IC07 image118b-red.jpg


We have used different materials:

  • DOPC, POPC, and Span 80 as surfactants
  • Dodecane and Mineral Oil as oils


We have done modifications to our protocol, often in an expedient manner.

  • Sometimes haphazard, sometimes serendipitous.


Pore Dilemma

IC07 runner index 4.png

IC07 scale.png
  • Vesicles are inefficient without permeability.
  • Pores do not destabilize the membrane, but they are toxic.
  • Detergents are not toxic, but they destabilize the membrane.


What's Next

IC07 runner index 5.png
  • Permeability Tests
  • Cell-free extract in-veso
  • Application-based experiments

IC07 veso-roadmap.png