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Team:Paris Bettencourt/Encapsulation
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==Experiments and results== | ==Experiments and results== | ||
| - | === | + | ===Cell Containment Assay=== |
| + | Our objective is to entrap cells that are still viable and able to perform metabolism. To asses this, beads were suspended in buffer and allowed to incubate at room temperature over several days. Presuming that treated beads could result in total cell containment, we wished to see if more viable cells would be released by physically destroying the beads. | ||
====Experimental setup==== | ====Experimental setup==== | ||
| - | + | * 2% Alginate beads containing cells were prepared (50mL saturated culture resuspended in 15 mL fresh LB and mixed with 15 mL 4% Alginate). | |
| + | |||
| + | * 4g beads were set aside in PBS at 4° as a negative control for containment (untreated alginate). | ||
| + | |||
| + | * 8g beads were treated as described above with polyethyleneimine and glutaraldehyde. | ||
| + | |||
| + | * 4g of treated beads were broken by cutting with a razor blade | ||
| + | |||
| + | * 4g of Untreated, Treated, and Treated & Broken beads were suspended in PBS buffer and left at room temperature. | ||
| + | |||
| + | * 100μL of supernatant was plated periodically to quantify release of cells. | ||
| + | |||
====Results==== | ====Results==== | ||
Revision as of 19:10, 24 September 2012
- Main
- Team
- Project
- Achievements
- Human Practice
- Safety
- Notebook
- Attributions
Overview
Delay system
Semantic containment
Restriction enzyme system
MAGE
Suicide system
Encapsulation
Synthetic import domain
Safety Questions
Safety AssessmentContents |
Overview
Provide an overview of this sub-project in a couple of phrases
Objectives
Our goal is to design a live-bacteria entrapment system. More than just encapsulating bacteria, we want to fully prevent their escape from the bead body into the surroundings. Alginate and other gel-based beads have been used successfully to prolong enzymatic activity in bioreactors[REF], but systems such as these are designed to allow steady release of microbes.
Design
Experiments and results
Cell Containment Assay
Our objective is to entrap cells that are still viable and able to perform metabolism. To asses this, beads were suspended in buffer and allowed to incubate at room temperature over several days. Presuming that treated beads could result in total cell containment, we wished to see if more viable cells would be released by physically destroying the beads.
Experimental setup
- 2% Alginate beads containing cells were prepared (50mL saturated culture resuspended in 15 mL fresh LB and mixed with 15 mL 4% Alginate).
- 4g beads were set aside in PBS at 4° as a negative control for containment (untreated alginate).
- 8g beads were treated as described above with polyethyleneimine and glutaraldehyde.
- 4g of treated beads were broken by cutting with a razor blade
- 4g of Untreated, Treated, and Treated & Broken beads were suspended in PBS buffer and left at room temperature.
- 100μL of supernatant was plated periodically to quantify release of cells.
Results
Present your results
Testing of the system
Experimental setup
Describe the experiment
Results
Present your results
