iLCD Reproduce the chemical activation of Calcium channels in - PowerPoint PPT Presentation

iGEM Valencia Lightning Cell Display iLCD

Reproduce the chemical activation of Calcium channels in yeast. Design a model for the electrical activation. Construction of device to supply the initial conditions of the characterization experiments. Characterization of the system parameters.

Luminometer

30 μ L KOH 100 Luminiscence ¡(a.u.) 80 60 40 20 0 0 50 100 150 200 Time ¡(s)

Characterization. Different volumes of KOH ( μ L)

Controls. Alkali shock Cch1: Calcium channel deficient mutant EDTA: Ethylenediaminetetraacetic acid (calcium quelant) Mid1: Calcium channel deficient mutant KCl: Potassium chloride WT: Wild Type

Controls. Alkali shock Channel deficient mutants Cch1: Calcium channel deficient mutant EDTA: Ethylenediaminetetraacetic acid (calcium quelant) Mid1: Calcium channel deficient mutant KCl: Potassium chloride WT: Wild Type

Device Voltage amplitude: ±0’1V During time intervals from 20 ms

4,5 Volts during 5 seconds

4,5 Volts during 5 seconds

Controls. Electrical Shock. WT: Wild Type SD + Coel: Medium with no yeasts EDTA: Ethylenediaminetetraacetic acid incubated with coelenterazine. (calcium quelant) Cch1: Calcium channel deficient mutant SD - Coel: Medium with yeasts Mid1: Calcium channel deficient mutant incubated without coelenterazine.

Most important parameters • Quantity of light emission • Reproduibility • Refreshing time

Characterization. Different time, same voltage Dead yeasts :( 1s 5s 10s

4,5 Volts during 5 seconds WT: Wild Type WT - coe: Wild Type incubated without coelenterazine

Device Voltage amplitude ±0’1V During time intervals from 20 ms 96 outputs

VIDEO JUNY

VIDEO JUNY

Reproduce the chemical activation of Calcium channels in yeast. Design a model for the electrical activation of these channels which points to a feasible activation at acceptable voltage levels. Construction of a homemade device to supply, very precisely, the initial conditions for the experiments. Mesurements and precise characterization of LECs (Lighting Emiting Cell)

Reproduce the chemical activation of Calcium channels in yeast. Design a model for the electrical activation of these channels which points to a feasible activation at acceptable voltage levels. Construction of a homemade device to supply, very precisely, the initial conditions for the experiments. Mesurements and precise characterization of LECs (Lighting Emiting Cell) Construction of a labview program which allows the cooperative work of several cultures to build the iLCD!!!

Achievements: • Electrical induction for a desired cell behaviour

Achievements: • Electrical induction for a desired cell behaviour • Characterization and construction of the first bio-screen of the history

Achievements: • Electrical induction for a desired cell behaviour • Characterization and construction of the first bio-screen of the history • BBa_K222000 (Aequorin) + Full characterization

Achievements: • Electrical induction for a desired cell behaviour • Characterization and construction of the first bio-screen of the history • BBa_K222000 (Aequorin) + Full characterization • Human Practices Report First review of iGEM and Human Practices (2005-2009) Biggest survey ever made (1288 people)